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  • Product Name
    Phospho-ERK1/2 (Thr202/Tyr204) Antibody
  • Catalog No.
    AF1015
  • Source
    Rabbit
  • Application
    WB,IHC,IF/ICC,ELISA
  • Reactivity
    Hm,Ms,Rt
  • UniProt
  • Mol.Wt.
    42,44kDa
  • Concentration
    1mg/ml
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Product Information

Alternative Names:Expand▼

ERK 1; ERK; ERK-1; ERK1; ERT 2; ERT2; Extracellular Signal Regulated Kinase 1; Extracellular signal related kinase 1; Extracellular signal-regulated kinase 1; HGNC6877; HS44KDAP; HUMKER1A; Insulin Stimulated MAP2 Kinase; Insulin-stimulated MAP2 kinase; MAP kinase 1; MAP kinase 3; MAP Kinase; MAP kinase isoform p44; MAPK 1; MAPK 3; MAPK; MAPK1; Mapk3; MGC20180; Microtubule Associated Protein 2 Kinase; Microtubule-associated protein 2 kinase; Mitogen Activated Protein Kinase 3; Mitogen-activated protein kinase 1; Mitogen-activated protein kinase 3; MK03_HUMAN; OTTHUMP00000174538; OTTHUMP00000174541; p44 ERK1; p44 MAPK; p44-ERK1; p44-MAPK; P44ERK1; P44MAPK; PRKM 3; PRKM3; Protein Kinase Mitogen Activated 3; ERK 2; ERK; ERK-2; ERT1; Extracellular Signal Regulated Kinase 2; Extracellular signal-regulated kinase 2; MAP kinase 1; MAP kinase 2; MAP kinase isoform p42; MAPK 1; MAPK 2; Mapk1; MAPK2; Mitogen-activated protein kinase 1; Mitogen-activated protein kinase 2; MK01_HUMAN; P38; P40; P41; p42-MAPK; P42MAPK; PRKM1; PRKM2; protein kinase, mitogen-activated, 1; protein kinase, mitogen-activated, 2; protein tyrosine kinase ERK2;

Applications:

WB 1:500-1:2000 IHC 1:50-1:200 IF 1:200, ELISA(peptide) 1:20000-1:40000

Reactivity:

Human,Mouse,Rat

Source:

Rabbit

Clonality:

Polyclonal

Purification:

The antibody is from purified rabbit serum by affinity purification via sequential chromatography on phospho- and non-phospho-peptide affinity columns.

Specificity:

Phospho-ERK1/2 (Thr202/Tyr204) Antibody detects endogenous levels of ERK1/2 only when phosphorylated at Threonine 202/Tyrosine 204.

Format:

Liquid

Concentration:

1mg/ml

Storage Condition and Buffer:

Rabbit IgG in phosphate buffered saline , pH 7.4, 150mM NaCl, 0.02% sodium azide and 50% glycerol.Store at -20 °C.Stable for 12 months from date of receipt.

Immunogen Information

Immunogen:

A synthesized peptide derived from human ERK1/2 around the phosphorylation site of Threonine 202/Tyrosine 204.

Uniprot:



>>Visit The Human Protein Atlas

Gene id:

Molecular Weight:

Observed Mol.Wt.: 42,44kDa.
Predicted Mol.Wt.: 44kDa.

Subcellular Location:

Nucleus.

Description:

Serine/threonine kinase which acts as an essential component of the MAP kinase signal transduction pathway. MAPK1/ERK2 and MAPK3/ERK1 are the 2 MAPKs which play an important role in the MAPK/ERK cascade. They participate also in a signaling cascade initiated by activated KIT and KITLG/SCF. Depending on the cellular context, the MAPK/ERK cascade mediates diverse biological functions such as cell growth, adhesion, survival and differentiation through the regulation of transcription, translation, cytoskeletal rearrangements. Publishing reference https://www.spandidos-publications.com/10.3892/mmr.2016.5305/download http://www.sciencedirect.com/science/article/pii/S0306452216000464 http://www.sciencedirect.com/science/article/pii/S0171298514002782 https://www.spandidos-publications.com/etm/12/1/499/download

Sequence:
        10         20         30         40         50
MAAAAAQGGG GGEPRRTEGV GPGVPGEVEM VKGQPFDVGP RYTQLQYIGE
60 70 80 90 100
GAYGMVSSAY DHVRKTRVAI KKISPFEHQT YCQRTLREIQ ILLRFRHENV
110 120 130 140 150
IGIRDILRAS TLEAMRDVYI VQDLMETDLY KLLKSQQLSN DHICYFLYQI
160 170 180 190 200
LRGLKYIHSA NVLHRDLKPS NLLINTTCDL KICDFGLARI ADPEHDHTGF
210 220 230 240 250
LTEYVATRWY RAPEIMLNSK GYTKSIDIWS VGCILAEMLS NRPIFPGKHY
260 270 280 290 300
LDQLNHILGI LGSPSQEDLN CIINMKARNY LQSLPSKTKV AWAKLFPKSD
310 320 330 340 350
SKALDLLDRM LTFNPNKRIT VEEALAHPYL EQYYDPTDEP VAEEPFTFAM
360 370
ELDDLPKERL KELIFQETAR FQPGVLEAP

Background

Function:

Serine/threonine kinase which acts as an essential component of the MAP kinase signal transduction pathway. MAPK1/ERK2 and MAPK3/ERK1 are the 2 MAPKs which play an important role in the MAPK/ERK cascade. They participate also in a signaling cascade initiated by activated KIT and KITLG/SCF. Depending on the cellular context, the MAPK/ERK cascade mediates diverse biological functions such as cell growth, adhesion, survival and differentiation through the regulation of transcription, translation, cytoskeletal rearrangements. The MAPK/ERK cascade plays also a role in initiation and regulation of meiosis, mitosis, and postmitotic functions in differentiated cells by phosphorylating a number of transcription factors. About 160 substrates have already been discovered for ERKs. Many of these substrates are localized in the nucleus, and seem to participate in the regulation of transcription upon stimulation. However, other substrates are found in the cytosol as well as in other cellular organelles, and those are responsible for processes such as translation, mitosis and apoptosis. Moreover, the MAPK/ERK cascade is also involved in the regulation of the endosomal dynamics, including lysosome processing and endosome cycling through the perinuclear recycling compartment (PNRC); as well as in the fragmentation of the Golgi apparatus during mitosis. The substrates include transcription factors (such as ATF2, BCL6, ELK1, ERF, FOS, HSF4 or SPZ1), cytoskeletal elements (such as CANX, CTTN, GJA1, MAP2, MAPT, PXN, SORBS3 or STMN1), regulators of apoptosis (such as BAD, BTG2, CASP9, DAPK1, IER3, MCL1 or PPARG), regulators of translation (such as EIF4EBP1) and a variety of other signaling-related molecules (like ARHGEF2, FRS2 or GRB10). Protein kinases (such as RAF1, RPS6KA1/RSK1, RPS6KA3/RSK2, RPS6KA2/RSK3, RPS6KA6/RSK4, SYK, MKNK1/MNK1, MKNK2/MNK2, RPS6KA5/MSK1, RPS6KA4/MSK2, MAPKAPK3 or MAPKAPK5) and phosphatases (such as DUSP1, DUSP4, DUSP6 or DUSP16) are other substrates which enable the propagation the MAPK/ERK signal to additional cytosolic and nuclear targets, thereby extending the specificity of the cascade.

Post-translational Modifications:

Phosphorylated upon KIT and FLT3 signaling (By similarity). Dually phosphorylated on Thr-202 and Tyr-204, which activates the enzyme. Ligand-activated ALK induces tyrosine phosphorylation. Dephosphorylated by PTPRJ at Tyr-204.

Subcellular Location:

Nucleus;

Extracellular region or secreted Cytosol Plasma membrane Cytoskeleton Lysosome Endosome Peroxisome ER Golgi apparatus Nucleus Mitochondrion Manual annotation Automatic computational assertionGraphics by Christian Stolte

Subunit Structure:

Binds both upstream activators and downstream substrates in multimolecular complexes. Found in a complex with at least BRAF, HRAS, MAP2K1/MEK1, MAPK3 and RGS14 (By similarity). Binds to HIV-1 Nef through its SH3 domain. This interaction inhibits its tyrosine-kinase activity. Interacts with ADAM15, ARRB2, CANX, DAPK1 (via death domain), HSF4, IER3, MAP2K1/MEK1, MORG1, NISCH, and SGK1. Interacts with PEA15 and MKNK2 (By similarity). MKNK2 isoform 1 binding prevents from dephosphorylation and inactivation (By similarity). Interacts with TPR. Interacts with CDKN2AIP. Interacts with HSF1 (via D domain and preferentially with hyperphosphorylated form); this interaction occurs upon heat shock (PubMed:10747973). Interacts with CAVIN4 (By similarity).

Similarity:

The TXY motif contains the threonine and tyrosine residues whose phosphorylation activates the MAP kinases.Belongs to the protein kinase superfamily. CMGC Ser/Thr protein kinase family. MAP kinase subfamily.

Research Fields

Research Fields:

· Cellular Processes > Cellular community - eukaryotes > Focal adhesion.(View pathway)
· Cellular Processes > Cell growth and death > Oocyte meiosis.(View pathway)
· Cellular Processes > Cellular community - eukaryotes > Gap junction.(View pathway)
· Cellular Processes > Cell growth and death > Apoptosis.(View pathway)
· Cellular Processes > Transport and catabolism > Autophagy - animal.(View pathway)
· Cellular Processes > Cellular community - eukaryotes > Adherens junction.(View pathway)
· Cellular Processes > Cellular community - eukaryotes > Signaling pathways regulating pluripotency of stem cells.(View pathway)
· Cellular Processes > Cell growth and death > Cellular senescence.(View pathway)
· Cellular Processes > Cell motility > Regulation of actin cytoskeleton.(View pathway)
· Environmental Information Processing > Signal transduction > TNF signaling pathway.(View pathway)
· Environmental Information Processing > Signal transduction > MAPK signaling pathway.(View pathway)
· Environmental Information Processing > Signal transduction > ErbB signaling pathway.(View pathway)
· Environmental Information Processing > Signal transduction > Sphingolipid signaling pathway.(View pathway)
· Environmental Information Processing > Signal transduction > FoxO signaling pathway.(View pathway)
· Environmental Information Processing > Signal transduction > mTOR signaling pathway.(View pathway)
· Environmental Information Processing > Signal transduction > Apelin signaling pathway.(View pathway)
· Environmental Information Processing > Signal transduction > cGMP-PKG signaling pathway.(View pathway)
· Environmental Information Processing > Signal transduction > Ras signaling pathway.(View pathway)
· Environmental Information Processing > Signal transduction > TGF-beta signaling pathway.(View pathway)
· Environmental Information Processing > Signal transduction > HIF-1 signaling pathway.(View pathway)
· Environmental Information Processing > Signal transduction > Rap1 signaling pathway.(View pathway)
· Environmental Information Processing > Signal transduction > Phospholipase D signaling pathway.(View pathway)
· Environmental Information Processing > Signal transduction > PI3K-Akt signaling pathway.(View pathway)
· Environmental Information Processing > Signal transduction > cAMP signaling pathway.(View pathway)
· Human Diseases > Cancers: Specific types > Pancreatic cancer.(View pathway)
· Human Diseases > Cancers: Specific types > Renal cell carcinoma.(View pathway)
· Human Diseases > Cancers: Specific types > Thyroid cancer.(View pathway)
· Human Diseases > Cancers: Specific types > Bladder cancer.(View pathway)
· Human Diseases > Cancers: Overview > Pathways in cancer.(View pathway)
· Human Diseases > Cancers: Specific types > Gastric cancer.(View pathway)
· Human Diseases > Cancers: Specific types > Non-small cell lung cancer.(View pathway)
· Human Diseases > Cancers: Specific types > Glioma.(View pathway)
· Human Diseases > Cancers: Specific types > Colorectal cancer.(View pathway)
· Human Diseases > Cancers: Overview > Proteoglycans in cancer.
· Human Diseases > Cancers: Specific types > Hepatocellular carcinoma.(View pathway)
· Human Diseases > Cancers: Specific types > Acute myeloid leukemia.(View pathway)
· Human Diseases > Cancers: Specific types > Endometrial cancer.(View pathway)
· Human Diseases > Endocrine and metabolic diseases > Type II diabetes mellitus.
· Human Diseases > Cancers: Specific types > Breast cancer.(View pathway)
· Human Diseases > Cancers: Specific types > Chronic myeloid leukemia.(View pathway)
· Human Diseases > Infectious diseases: Bacterial > Pertussis.
· Human Diseases > Infectious diseases: Bacterial > Salmonella infection.
· Human Diseases > Cancers: Overview > Central carbon metabolism in cancer.(View pathway)
· Human Diseases > Cancers: Overview > Choline metabolism in cancer.(View pathway)
· Human Diseases > Infectious diseases: Bacterial > Shigellosis.
· Human Diseases > Infectious diseases: Viral > Hepatitis C.
· Human Diseases > Cancers: Overview > Viral carcinogenesis.
· Human Diseases > Infectious diseases: Viral > Hepatitis B.
· Human Diseases > Cancers: Specific types > Melanoma.(View pathway)
· Human Diseases > Infectious diseases: Viral > Human papillomavirus infection.
· Human Diseases > Infectious diseases: Parasitic > Chagas disease (American trypanosomiasis).
· Human Diseases > Drug resistance: Antineoplastic > EGFR tyrosine kinase inhibitor resistance.
· Human Diseases > Infectious diseases: Bacterial > Tuberculosis.
· Human Diseases > Infectious diseases: Viral > Influenza A.
· Human Diseases > Drug resistance: Antineoplastic > Platinum drug resistance.
· Human Diseases > Infectious diseases: Parasitic > Leishmaniasis.
· Human Diseases > Substance dependence > Alcoholism.
· Human Diseases > Neurodegenerative diseases > Prion diseases.
· Human Diseases > Neurodegenerative diseases > Alzheimer's disease.
· Human Diseases > Infectious diseases: Parasitic > Toxoplasmosis.
· Human Diseases > Drug resistance: Antineoplastic > Endocrine resistance.
· Human Diseases > Cancers: Specific types > Prostate cancer.(View pathway)
· Organismal Systems > Immune system > Th1 and Th2 cell differentiation.(View pathway)
· Organismal Systems > Immune system > Toll-like receptor signaling pathway.(View pathway)
· Organismal Systems > Immune system > IL-17 signaling pathway.(View pathway)
· Organismal Systems > Immune system > Platelet activation.(View pathway)
· Organismal Systems > Immune system > T cell receptor signaling pathway.(View pathway)
· Organismal Systems > Immune system > Fc epsilon RI signaling pathway.(View pathway)
· Organismal Systems > Immune system > Fc gamma R-mediated phagocytosis.(View pathway)
· Organismal Systems > Endocrine system > Progesterone-mediated oocyte maturation.
· Organismal Systems > Immune system > Natural killer cell mediated cytotoxicity.(View pathway)
· Organismal Systems > Endocrine system > Relaxin signaling pathway.
· Organismal Systems > Endocrine system > Thyroid hormone signaling pathway.(View pathway)
· Organismal Systems > Circulatory system > Adrenergic signaling in cardiomyocytes.(View pathway)
· Organismal Systems > Excretory system > Aldosterone-regulated sodium reabsorption.
· Organismal Systems > Endocrine system > Insulin signaling pathway.(View pathway)
· Organismal Systems > Immune system > Th17 cell differentiation.(View pathway)
· Organismal Systems > Endocrine system > Prolactin signaling pathway.(View pathway)
· Organismal Systems > Nervous system > Long-term potentiation.
· Organismal Systems > Nervous system > Neurotrophin signaling pathway.(View pathway)
· Organismal Systems > Circulatory system > Vascular smooth muscle contraction.(View pathway)
· Organismal Systems > Development > Osteoclast differentiation.(View pathway)
· Organismal Systems > Endocrine system > Oxytocin signaling pathway.
· Organismal Systems > Development > Axon guidance.(View pathway)
· Organismal Systems > Nervous system > Long-term depression.
· Organismal Systems > Nervous system > Glutamatergic synapse.
· Organismal Systems > Nervous system > Cholinergic synapse.
· Organismal Systems > Environmental adaptation > Circadian entrainment.
· Organismal Systems > Immune system > NOD-like receptor signaling pathway.(View pathway)
· Organismal Systems > Endocrine system > Melanogenesis.
· Organismal Systems > Immune system > Chemokine signaling pathway.(View pathway)
· Organismal Systems > Nervous system > Retrograde endocannabinoid signaling.(View pathway)
· Organismal Systems > Nervous system > Serotonergic synapse.
· Organismal Systems > Immune system > B cell receptor signaling pathway.(View pathway)
· Organismal Systems > Endocrine system > Estrogen signaling pathway.(View pathway)

FCN‐A/2, acting as a new regulator of macrophage polarization, mediates the inflammatory response in experimental mouse colitis YF Yang, YD Zhou, JC Hu, FL Luo, Y Xie… …, 2017 Wiley Online Library
This image is a courtesy of anonymous review
This image is a courtesy of anonymous review
Western blot analysis of ERK1/2 phosphorylation expression in HeLa whole cell lysates,The lane on the left is treated with the antigen-specific peptide.
This image is a courtesy of anonymous review
AF1015 at 1/200 staining Rat lung tissue sections by IHC-P. The tissue was formaldehyde fixed and a heat mediated antigen retrieval step in citrate buffer was performed. The tissue was then blocked and incubated with the antibody for 1.5 hours at 22°C. An HRP conjugated goat anti-rabbit antibody was used as the secondary.
AF1015 at 1/200 staining Rat lung tissue sections by IHC-P. The tissue was formaldehyde fixed and a heat mediated antigen retrieval step in citrate buffer was performed. The tissue was then blocked and incubated with the antibody for 1.5 hours at 22°C. An HRP conjugated goat anti-rabbit antibody was used as the secondary.
AF1015 at 1/200 staining Mouse liver tissue sections by IHC-P. The tissue was formaldehyde fixed and a heat mediated antigen retrieval step in citrate buffer was performed. The tissue was then blocked and incubated with the antibody for 1.5 hours at 22°C. An HRP conjugated goat anti-rabbit antibody was used as the secondary.
AF1015 at 1/200 staining Human pancreas tissue sections by IHC-P. The tissue was formaldehyde fixed and a heat mediated antigen retrieval step in citrate buffer was performed. The tissue was then blocked and incubated with the antibody for 1.5 hours at 22°C. An HRP conjugated goat anti-rabbit antibody was used as the secondary.
AF1015 at 1/200 staining Human heart tissue sections by IHC-P. The tissue was formaldehyde fixed and a heat mediated antigen retrieval step in citrate buffer was performed. The tissue was then blocked and incubated with the antibody for 1.5 hours at 22°C. An HRP conjugated goat anti-rabbit antibody was used as the secondary.
AF1015 at 1/200 staining Human heart tissue sections by IHC-P. The tissue was formaldehyde fixed and a heat mediated antigen retrieval step in citrate buffer was performed. The tissue was then blocked and incubated with the antibody for 1.5 hours at 22°C. An HRP conjugated goat anti-rabbit antibody was used as the secondary.
Phospho-ERK1/2 (Thr202/Tyr204) Antibody for IHC in human testis
AF1015 staining lovo cells by ICC/IF. Cells were fixed with PFA and permeabilized in 0.1% saponin prior to blocking in 10% serum for 45 minutes at 37°C. The primary antibody was diluted 1/400 and incubated with the sample for 1 hour at 37°C. A Alexa Fluor® 594 conjugated goat polyclonal to rabbit IgG (H+L), diluted 1/600 was used as secondary antibody.
ELISA analysis of AF1015 showing specificity to Phospho-ERK1/2 (Thr202/Tyr204) peptide. Peptides concentration: 1ug/ml.
P-peptide: phospho-peptide; N-peptide: non-phospho-peptide.

Reference Citations:

1). Jin X et al. Tiliroside, the major component of Agrimonia pilosa Ledeb ethanol extract, inhibits MAPK/JNK/p38-mediated inflammation in lipopolysaccharide-activated RAW 264.7 macrophages. Exp Ther Med 2016 Jul;12(1):499-505 (PubMed: 27347085)

Application: WB    Species: mouse;    Sample:Not available;

Figure 7. Effect of tiliroside on the phosphorylation of mitogen‑activated protein kinase‑ERK/JNK/p38 proteins. (A) RAW 264.7 cells were treated with LPS 1 µg/ml with or without tiliroside (12.5, 25, 50, and 100 µM) for 45 min and the expression of p‑ERK1/2, p‑JNK and p‑p38 was assessed by western blot analysis. Detection of β‑actin was conducted to confirm the equal loading of proteins. Densitometric analysis of p‑ERK1/2 (B) p‑JNK (C) and p‑p38 (D) expression represent the mean±standard deviation of three separate experiments. Data were normalized with respect to β‑actin levels. **P


2). Jin X et al. 11-O-acetylcyathatriol inhibits MAPK/p38-mediated inflammation in LPS-activated RAW 264.7 macrophages and has a protective effect on ethanol-induced gastric injury. Mol Med Rep 2016 Jul;14(1):874-80 (PubMed: 27222252)

Application: WB    Species: mouse;    Sample:Not available;

Figure 3. Effects of 11‑O‑acetylcyathatriol on the protein expression levels of iNOS and COX‑2. The RAW 264.7 cells were treated by 1 µg/ml of LPS with indicated concentrations of 11-O‑acetylcyathatriol (12.5, 25, 50 and 100 µM) for 24 h, and the expression levels of (A) iNOS and COX‑2 were detected using western blot analysis. (B) Effects of 11‑O-acetylcyathatriol on the phosphorylation of ERK1/2, JNK and p38 proteins. RAW 264.7 cells were treated with 1 µg/ml LPS with 11‑O‑acetylcyathatriol (12.5, 25, 50 and 100 µM) for 30 min, and the protein expression levels of p‑ERK1/2, p‑JNK and p‑p38 were detected using western blot analysis. (C) Effects of 11‑O-acetylcyathatriol on the protein degradation of IκB-α. RAW 264.7 cells were treated with 1 µg/ml of LPS with 11-O‑acetylcyathatriol (12.5, 25, 50 and 100 µM) for 10 min, and the protein expression of IκB-α was detected using western blot analysis. iNOS, inducible nitric oxide synthase; COX‑2, cyclooxygenase‑2; LPS, lipopolysaccharide; ERK, extracellular signal‑regulated kinase; JNK, c‑Jun N‑terminal kinase; IκB-α, inhibitor of nuclear factor-κB-α; p‑, phosphorylated.


3). Su Z et al. HMGB1 modulates Lewis cell autophagy and promotes cell survival via RAGE-HMGB1-Erk1/2 positive feedback during nutrient depletion. Immunobiology 2015 May;220(5):539-44 (PubMed: 25578401)

Application: WB    Species:Not available;    Sample:Not available;


4). Huang C et al. Estrogen regulates excitatory amino acid carrier 1 (EAAC1) expression through sphingosine kinase 1 (SphK1) transacting FGFR-mediated ERK signaling in rat C6 astroglial cells. Neuroscience 2016 Apr 5;319:9-22 (PubMed: 26804240)

Application: WB 1/800    Species: rat;    Sample: rat;

Fig. 4. FGFR- ERK signaling pathway is involved in FGF2-induced EAAC1 expression. (A) C6 cells were incubated with FGF2 (10 ng/mL) for different durations. (B, C) Before incubation with FGF2 (10 ng/mL) for 15 min (B) or 24 h (C), cells were pretreated with the FGFR kinase inhibitor SU5402 (SU; 25 lM, 5 min) or the ERK inhibitor U0126 (U0; 10 lM, 1 h). Phosphorylation of FRS2a or ERK1/2 (A and B, respectively), and EAAC1 expression (C) were analyzed by Western blots. Data are shown as mean ± SEM from three independent experiments. * p < 0.05; **p < 0.01 versus the control group. # p < 0.05; ##p < 0.01 versus the FGF2 alone group.


5). Song T et al. GPR120 promotes adipogenesis through intracellular calcium and extracellular signal-regulated kinase 1/2 signal pathway. Mol Cell Endocrinol 2016 Oct 15;434:1-13 (PubMed: 27302893)

Application: WB    Species:Not available;    Sample:Not available;


6). Xie XC et al. Relaxin Attenuates Contrast-Induced Human Proximal Tubular Epithelial Cell Apoptosis by Activation of the PI3K/Akt Signaling Pathway In Vitro. Biomed Res Int 2017;2017:2869405 (PubMed: 28540295)

7). Deng LJ et al. Oxabicycloheptene Sulfonate Protects Against β-Amyloid-induced Toxicity by Activation of PI3K/Akt and ERK Signaling Pathways Via GPER1 in C6 Cells. Neurochem Res 2017 Aug;42(8):2246-2256 (PubMed: 28374135)

Application: WB 1/500    Species: rat;    Sample: C6 Cells;

Fig. 3 OBHS and RAL increase Akt and ERK phosphorylation through GPER1. a Akt and ERK inhibitors attenuated the pro-survival effects of OBHS and RAL against Aβ toxicity. Cells were pretreated with specific inhibitors LY294002 (LY, 20  μM), U0126 (U,10 μM) for one hour and then incubated with OBHS (1 μM) or RAL (1 μM) for 45 min, after Aβ treatment for an additional 24 h. Cell viability was measured by MTT method. b,c C6 cells were preincubated with PI3K/Akt inhibitor LY (b) or ERK inhibitor U (c) for one hour, followed by OBHS (1 μM) or RAL (1 μM) treatment for 15 min. d GPER1 antagonist G15 (10  μM) was added to the media for 24  h, before OBHS (1  μM) or RAL (1  μM) treatment for 15  min of C6 cells. The proteins were analyzed by the western blot method. The data were shown as the mean±SD from three independent experiments.


8). Li ZH et al. miR-29a regulated ER-positive breast cancer cell growth and invasion and is involved in the insulin signaling pathway. Oncotarget 2017 May 16;8(20):32566-32575 (PubMed: 28427228)

Application: WB    Species: human;    Sample: MCF-7;

Figure 5: Effect of miR-29a on the insulin signaling pathways. (A) IGF-1R, CDC42, p85α, p-ERK and ERK mRNA levels were determined by qPCR in ER-positive cells transfected with a miR-29a mimic or miR-29a inhibitor. (B, C) Western blotting analysis was performed to monitor IGF-1R, CDC42, p85α, p-ERK and ERK expression in ER-positive cells transfected with a miR-29a mimic or miR-29a inhibitor. The results showed that ERK, CDC42 and p85α are the target genes of miR-29a; however, miR-29a promotes breast cancer cell growth and proliferation mainly by activating ERK phosphorylation.


9). Cui C et al. Hypoxic postconditioning attenuates apoptosis via inactivation of adenosine A2a receptor through NDRG3-Raf-ERK pathway. Biochem Biophys Res Commun 2017 Sep 16;491(2):277-284 (PubMed: 28743501)

Application: WB 1/1000    Species: human;    Sample:Not available;

Fig.2 Protein Expression of NDRD3-Raf-ERK pathway, A2a receptors, Cytochrome C.


10). Peng J et al. MiR-377 promotes white adipose tissue inflammation and decreases insulin sensitivity in obesity via suppression of sirtuin-1 (SIRT1). Oncotarget 2017 Jul 31;8(41):70550-70563 (PubMed: 29050301)

Application: WB    Species: mouse;    Sample:Not available;

Figure 4: MiR-377 promotes inflammation and insulin-resistance in mature 3T3-L1 cells. After transfection with 100 nM miR-377 mimics or inhibitor for 24 h, differentiated 3T3-L1 adipocytes were treated with 10 ng/ml TNFα for 24 h and then stimulated with 100 nM insulin for 15 min. Cells were then harvested for real-time PCR and immunoblotting analyses. (A) The effect of miR-377 overexpression on inflammation-related gene expression. *P < 0.05, **P < 0.01 vs. NC 0.1% BSA; ##P < 0.01 vs. NC 10 ng/ml TNFα; ns, not signifcant (n = 3). (B) The effect of miR-377 inhibition on inflammation-related gene expression under conditions of TNFα-induced insulin-resistance. ##P < 0.01 vs. NC 10 ng/ml TNFα(n = 3). (C) The effect of miR-377 overexpression on JNK phosphorylation under conditions of TNFα-induced insulin-resistance. *P < 0.05 vs. NC without insulin; #P < 0.05 vs. NC with insulin (n = 3). (D and E) The effect of miR-377 overexpression on AKT and ERK phosphorylation. *P < 0.05 vs. NC with 0.1% BSA and insulin; #P < 0.05 vs. NC with 10 ng/ml TNFα and insulin (n = 3). (F) The effect of miR-377 inhibition on JNK phosphorylation under conditions of TNFα-induced insulin-resistance. *P < 0.05 vs. NC without insulin; #P < 0.05 vs. NC with insulin (n = 3). (G and H) The effect of miR-377 inhibition on AKT and ERK phosphorylation. *P < 0.05 vs. NC with 0.1% BSA and insulin; #P < 0.05 vs. NC with 10 ng/ml TNFα and insulin (n = 3).


11). Zhang X et al. Neuroprotective Effect of Modified Xijiao Dihuang Decoction against Oxygen-Glucose Deprivation and Reoxygenation-Induced Injury in PC12 Cells: Involvement of TLR4-MyD88/NF-κB Signaling Pathway. Evid Based Complement Alternat Med 2017;2017:3848595 (PubMed: 29234386)

12). Li H et al. Dysifragilone A inhibits LPS‑induced RAW264.7 macrophage activation by blocking the p38 MAPK signaling pathway. Mol Med Rep 2018 Jan;17(1):674-682 (PubMed: 29115475)

13). Liu X et al. MicroRNA-370 inhibits the growth and metastasis of lung cancer by down-regulating epidermal growth factor receptor expression. Oncotarget 2017 Oct 4;8(50):88139-88151 (PubMed: 29152147)

Application: WB 1/1000    Species: human;    Sample:Not available;

Figure 4: Induction of miR-370 over-expression reduces EGFR and HIF-1α expression and inhibits the ERK and AKT phosphorylation in XWLC-05 and H157 cells. XWLC-05 and H157 cells were transfected with miR-370 mimics, miR-370 inhibitor or corresponding controls for 24 h. The relative levels of EGFR, HIF-1α, ERK, AKT expression, ERK and AKT phosphorylation were determined by Western blot assays. Data are representative images or expressed as the means ± SEM of each group of cells from three separate experiments. *P<0.05, **P<0.01, ***P<0.001, ****P<0.0001.


14). Chai L et al. Biological functions of lung cancer cells are suppressed in co-culture with mesenchymal stem cells isolated from umbilical cord. Exp Ther Med 2018 Jan;15(1):1076-1080 (PubMed: 29399109)

15). Liu Y et al. LncRNA-TCONS_00034812 in cell proliferation and apoptosis of pulmonary artery smooth muscle cells and its mechanism. J Cell Physiol 2018 Jun;233(6):4801-4814 (PubMed: 29150946)

16). Wu Z et al. MFAP5 promotes tumor progression and bone metastasis by regulating ERK/MMP signaling pathways in breast cancer. Biochem Biophys Res Commun 2018 Apr 6;498(3):495-501 (PubMed: 29526753)

17). Liu Y et al. LncRNA-TCONS_00034812 in cell proliferation and apoptosis of pulmonary artery smooth muscle cells and its mechanism. J Cell Physiol 2018 Jun;233(6):4801-4814 (PubMed: 29150946)

18). Xin H et al. Chemokine CXCL3 mediates prostate cancer cells proliferation, migration and gene expression changes in an autocrine/paracrine fashion. Int Urol Nephrol 2018 May;50(5):861-868 (PubMed: 29524043)

19). Tang Q et al. Ferroptosis is newly characterized form of neuronal cell death in response to arsenite exposure. Neurotoxicology 2018 Jul;67:27-36 (PubMed: 29678591)

20). Huang YX et al. Ovostatin 2 knockdown significantly inhibits the growth, migration, and tumorigenicity of cutaneous malignant melanoma cells. PLoS One 2018 Apr 23;13(4):e0195610 (PubMed: 29684087)

21). Wang W et al. Umbilical cord‑derived mesenchymal stem cells can inhibit the biological functions of melanoma A375 cells. Oncol Rep 2018 Jul;40(1):511-517 (PubMed: 29767256)

22). Zhang Q et al. Anti-inflammatory action of ambuic acid, a natural product isolated from the solid culture of Pestalotiopsis neglecta, through blocking ERK/JNK mitogen-activated protein kinase signaling pathway. Exp Ther Med 2018 Aug;16(2):1538-1546 (PubMed: 30116402)

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Catalog Number :

AF1015-BP

Price/Size :

$200/1mg.
Tips: For phospho antibody, we provide phospho peptide(0.5mg) and non-phospho peptide(0.5mg).

Function :

Blocking peptides are peptides that bind specifically to the target antibody and block antibody binding. These peptide usually contains the epitope recognized by the antibody. Antibodies bound to the blocking peptide no longer bind to the epitope on the target protein. This mechanism is useful when non-specific binding is an issue, for example, in Western blotting (immunoblot) and immunohistochemistry (IHC). By comparing the staining from the blocked antibody versus the antibody alone, one can see which staining is specific; Specific binding will be absent from the western blot or immunostaining performed with the neutralized antibody.

Format and storage :

Synthetic peptide was lyophilized with 100% acetonitrile and is supplied as a powder. Reconstitute with 0.1 ml DI water for a final concentration of 1 mg/ml.The purity is >90%,tested by HPLC and MS.Storage Maintain refrigerated at 2-8°C for up to 6 months. For long term storage store at -20°C.

Precautions :

This product is for research use only. Not for use in diagnostic or therapeutic procedures.

IMPORTANT: For western blots, incubate membrane with diluted antibody in 5% w/v milk , 1X TBS, 0.1% Tween®20 at 4°C with gentle shaking, overnight.