Phospho-CREB (Ser133) Antibody | Affinity Biosciences

Product:	Phospho-CREB (Ser133) Antibody
Catalog:	AF3189
Source:	Rabbit
Application:	WB, IHC, IF/ICC
Reactivity:	Human, Mouse, Rat
Prediction:	Pig, Bovine, Horse, Sheep, Rabbit, Dog, Chicken, Xenopus
Mol.Wt.:	43kDa; 37kD(Calculated).
Uniprot:	P16220
RRID:	AB_2834621

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Cat.#S0018

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. By comparing the staining from the blocked antibody versus the antibody alone, one can see which staining is specific.

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100ul	$280	In stock
200ul	$350	In stock

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MSDS

Data Sheet

COA

Protocols

WB Handbook

IHC Protocol

IF/ICC Protocol

Product Info

Source:

Rabbit

Application:

WB 1:500-1:2000, IHC 1:100-1:500, IF/ICC 1:100-1:500
*The optimal dilutions should be determined by the end user.
*Tips:

WB: For western blot detection of denatured protein samples. IHC: For immunohistochemical detection of paraffin sections (IHC-p) or frozen sections (IHC-f) of tissue samples. IF/ICC: For immunofluorescence detection of cell samples. ELISA(peptide): For ELISA detection of antigenic peptide.

Reactivity:

Human,Mouse,Rat

Prediction:

Pig(100%), Bovine(100%), Horse(100%), Sheep(100%), Rabbit(100%), Dog(100%), Chicken(100%), Xenopus(100%)

Clonality:

Polyclonal

Specificity:

Phospho-CREB (Ser133) Antibody detects endogenous levels of CREB only when phosphorylated at Serine 133.

RRID:

AB_2834621
Cite Format: Affinity Biosciences Cat# AF3189, RRID:AB_2834621.

Conjugate:

Unconjugated.

Purification:

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

Storage:

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.

Alias:

Fold/Unfold

Active transcription factor CREB; cAMP response element binding protein 1; cAMP response element binding protein; cAMP responsive element binding protein 1; cAMP-responsive element-binding protein 1; CREB; CREB-1; CREB1; CREB1_HUMAN; Cyclic AMP-responsive element-binding protein 1; MGC9284; OTTHUMP00000163864; OTTHUMP00000163865; OTTHUMP00000206660; OTTHUMP00000206662; OTTHUMP00000206667; Transactivator protein;

Immunogens

Immunogen:

Uniprot:

P16220(CREB1_HUMAN) >>Visit HPA database.

Gene(ID):

CREB1(1385)

Description:

This gene encodes a transcription factor that is a member of the leucine zipper family of DNA binding proteins. This protein binds as a homodimer to the cAMP-responsive element, an octameric palindrome.

Sequence:

P16220 CREB1_HUMAN:
Protein BLAST With
NCBI/
ExPASy/
Uniprot

MTMESGAENQQSGDAAVTEAENQQMTVQAQPQIATLAQVSMPAAHATSSAPTVTLVQLPNGQTVQVHGVIQAAQPSVIQSPQVQTVQSSCKDLKRLFSGTQISTIAESEDSQESVDSVTDSQKRREILSRRPSYRKILNDLSSDAPGVPRIEEEKSEEETSAPAITTVTVPTPIYQTSSGQYIAITQGGAIQLANNGTDGVQGLQTLTMTNAAATQPGTTILQYAQTTDGQQILVPSNQVVVQAASGDVQTYQIRTAPTSTIAPGVVMASSPALPTQPAEEAARKREVRLMKNREAARECRRKKKEYVKCLENRVAVLENQNKTLIEELKALKDLYCHKSD

Predictions

Predictions:

Score>80(red) has high confidence and is suggested to be used for WB detection. *The prediction model is mainly based on the alignment of immunogen sequences, the results are for reference only, not as the basis of quality assurance.

Species

Results

Score

Pig

100

Horse

100

Bovine

100

Sheep

100

Dog

100

Xenopus

100

Chicken

100

Rabbit

100

Zebrafish

Model Confidence:
High(score>80) Medium(80>score>50) Low(score<50) No confidence

PTMs - P16220 As Substrate

P16220

Site	PTM Type	Enzyme	Source
S40	Phosphorylation		Uniprot
S80	Phosphorylation		Uniprot
S98	Phosphorylation	Q15139 (PRKD1)	Uniprot
T100	Phosphorylation	Q13315 (ATM)	Uniprot
S108	Phosphorylation	P68400 (CSNK2A1) , P48729 (CSNK1A1)	Uniprot
S111	Phosphorylation	P68400 (CSNK2A1) , P48729 (CSNK1A1) , Q13315 (ATM)	Uniprot
S114	Phosphorylation	P48729 (CSNK1A1)	Uniprot
S117	Phosphorylation		Uniprot
T119	Phosphorylation		Uniprot
S121	Phosphorylation	Q13535 (ATR) , Q13315 (ATM)	Uniprot
S129	Phosphorylation	P49841 (GSK3B)	Uniprot
S133	Phosphorylation	Q16539 (MAPK14) , P31751 (AKT2) , Q02156 (PRKCE) , O43781 (DYRK3) , Q96NX5 (CAMK1G) , Q16566 (CAMK4) , P17612 (PRKACA) , Q6SA08 (TSSK4) , Q99986 (VRK1) , Q13557 (CAMK2D) , Q15418 (RPS6KA1) , O75676 (RPS6KA4) , P28482 (MAPK1) , Q14012 (CAMK1) , Q06187 (BTK) , P51812 (RPS6KA3) , Q13627 (DYRK1A) , O75582 (RPS6KA5) , P41279 (MAP3K8) , O00141 (SGK1) , P27361 (MAPK3) , P31749 (AKT1) , Q13315 (ATM) , Q15349 (RPS6KA2) , Q9Y243 (AKT3) , P49137 (MAPKAPK2) , Q15139 (PRKD1)	Uniprot
Y134	Phosphorylation		Uniprot
K136	Acetylation		Uniprot
K136	Ubiquitination		Uniprot
S142	Phosphorylation	Q9UQM7 (CAMK2A) , Q13557 (CAMK2D)	Uniprot
S143	Phosphorylation		Uniprot
S156	Phosphorylation	P48729 (CSNK1A1)	Uniprot
T256	Phosphorylation		Uniprot
S270	Phosphorylation	P06493 (CDK1)	Uniprot
S271	Phosphorylation	Q9H2X6 (HIPK2) , P06493 (CDK1)	Uniprot
K285	Sumoylation		Uniprot
K304	Sumoylation		Uniprot
K309	Ubiquitination		Uniprot
R314	Methylation		Uniprot
K323	Ubiquitination		Uniprot
K330	Ubiquitination		Uniprot
K333	Ubiquitination		Uniprot
K339	Ubiquitination		Uniprot
S340	Phosphorylation		Uniprot

Research Backgrounds

P16220_CREB1_HUMAN

Function:

Phosphorylation-dependent transcription factor that stimulates transcription upon binding to the DNA cAMP response element (CRE), a sequence present in many viral and cellular promoters. Transcription activation is enhanced by the TORC coactivators which act independently of Ser-133 phosphorylation. Involved in different cellular processes including the synchronization of circadian rhythmicity and the differentiation of adipose cells.

PTMs:

Stimulated by phosphorylation. Phosphorylation of both Ser-133 and Ser-142 in the SCN regulates the activity of CREB and participates in circadian rhythm generation. Phosphorylation of Ser-133 allows CREBBP binding. In liver, phosphorylation is induced by fasting or glucagon in a circadian fashion (By similarity). CREBL2 positively regulates phosphorylation at Ser-133 thereby stimulating CREB1 transcriptional activity (By similarity). Phosphorylated upon calcium influx by CaMK4 and CaMK2 on Ser-133. CaMK4 is much more potent than CaMK2 in activating CREB. Phosphorylated by CaMK2 on Ser-142. Phosphorylation of Ser-142 blocks CREB-mediated transcription even when Ser-133 is phosphorylated. Phosphorylated by CaMK1 (By similarity). Phosphorylation of Ser-271 by HIPK2 in response to genotoxic stress promotes CREB1 activity, facilitating the recruitment of the coactivator CBP. Phosphorylated at Ser-133 by RPS6KA3, RPS6KA4 and RPS6KA5 in response to mitogenic or stress stimuli. Phosphorylated by TSSK4 on Ser-133.

Sumoylated with SUMO1. Sumoylation on Lys-304, but not on Lys-285, is required for nuclear localization of this protein. Sumoylation is enhanced under hypoxia, promoting nuclear localization and stabilization.

Subcellular Location:

Nucleus.

Subunit Structure:

Interacts with PPRC1. Binds DNA as a dimer. This dimer is stabilized by magnesium ions. Interacts, through the bZIP domain, with the coactivators TORC1/CRTC1, TORC2/CRTC2 and TORC3/CRTC3. When phosphorylated on Ser-133, binds CREBBP (By similarity). Interacts with CREBL2; regulates CREB1 phosphorylation, stability and transcriptional activity (By similarity). Interacts (phosphorylated form) with TOX3. Interacts with ARRB1. Binds to HIPK2. Interacts with SGK1. Interacts with TSSK4; this interaction facilitates phosphorylation on Ser-133. Forms a complex with KMT2A and CREBBP.

(Microbial infection) Interacts with hepatitis B virus/HBV protein X.

(Microbial infection) Interacts with HTLV-1 protein Tax.

Family&Domains:

Belongs to the bZIP family.

Research Fields

· Environmental Information Processing > Signal transduction > cGMP-PKG signaling pathway. (View pathway)

· Environmental Information Processing > Signal transduction > cAMP signaling pathway. (View pathway)

· Environmental Information Processing > Signal transduction > PI3K-Akt signaling pathway. (View pathway)

· Environmental Information Processing > Signal transduction > AMPK signaling pathway. (View pathway)

· Environmental Information Processing > Signal transduction > TNF signaling pathway. (View pathway)

· Human Diseases > Endocrine and metabolic diseases > Insulin resistance.

· Human Diseases > Neurodegenerative diseases > Huntington's disease.

· Human Diseases > Substance dependence > Cocaine addiction.

· Human Diseases > Substance dependence > Amphetamine addiction.

· Human Diseases > Substance dependence > Alcoholism.

· Human Diseases > Infectious diseases: Bacterial > Tuberculosis.

· Human Diseases > Infectious diseases: Viral > Hepatitis B.

· Human Diseases > Infectious diseases: Viral > Human papillomavirus infection.

· Human Diseases > Infectious diseases: Viral > HTLV-I infection.

· Human Diseases > Cancers: Overview > Viral carcinogenesis.

· Human Diseases > Cancers: Specific types > Prostate cancer. (View pathway)

· Organismal Systems > Aging > Longevity regulating pathway. (View pathway)

· Organismal Systems > Circulatory system > Adrenergic signaling in cardiomyocytes. (View pathway)

· Organismal Systems > Development > Osteoclast differentiation. (View pathway)

· Organismal Systems > Immune system > Antigen processing and presentation. (View pathway)

· Organismal Systems > Environmental adaptation > Circadian rhythm. (View pathway)

· Organismal Systems > Environmental adaptation > Circadian entrainment.

· Organismal Systems > Nervous system > Cholinergic synapse.

· Organismal Systems > Nervous system > Dopaminergic synapse.

· Organismal Systems > Endocrine system > Insulin secretion. (View pathway)

· Organismal Systems > Endocrine system > Estrogen signaling pathway. (View pathway)

· Organismal Systems > Endocrine system > Melanogenesis.

· Organismal Systems > Endocrine system > Thyroid hormone synthesis.

· Organismal Systems > Endocrine system > Glucagon signaling pathway.

· Organismal Systems > Endocrine system > Renin secretion.

· Organismal Systems > Endocrine system > Aldosterone synthesis and secretion.

· Organismal Systems > Endocrine system > Relaxin signaling pathway.

· Organismal Systems > Excretory system > Vasopressin-regulated water reabsorption.

References

1). Peng Z et al. MiR-29b-3p aggravates NG108-15 cell apoptosis triggered by fluorine combined with aluminum. Ecotoxicol Environ Saf 2021 Aug 20;224:112658. (PubMed: 34425535) [IF=7.129]

Application: WB Species: Rat Sample: NG108–15 cells

Fig. 4. mRNA and protein expression of Dusp2, p38 MAPK, CREB and JNK in NG108–15 cells treated with F, Al and FA. (A/B) The graphs show the results of the statistical analysis of Dusp2, p38 MAPK, CREB and JNK mRNA expression levels based on qRT-PCR. (C/D/E/F/G/H/I) The graphs show signaling pathway protein expression in NG108–15 cells as assessed by Western blotting analysis. a P < 0.05, compared with the control group; b P < 0.05, compared with the F group; c P < 0.05, compared with the Al group (n = 6).

2). Qiao YL et al. Uncaria rhynchophylla ameliorates unpredictable chronic mild stress-induced depression in mice via activating 5-HT1A receptor: Insights from transcriptomics. Phytomedicine 2021 Jan;81:153436. (PubMed: 33360346) [IF=6.656]

Application: WB Species: mice Sample: hippocampus

Fig. 11. Effects of URE on 5-HT1A signaling pathway. (A) Western blotting analysis of 5-HT1A, BDNF, p-CREB, CREB, p-PKA, and PKA in hippocampus. (B-E) Quantitative data of 5-HT1A (B), BDNF (C), p-CREB/CREB (D), and p-PKA/PKA (E), data represent mean ± SEM (n = 3), ** p < 0.01, *** p < 0.001 vs. the control group; # p < 0.05, ## p < 0.01, ### p < 0.001 vs. the UCMS group. (F) The agonistic effect of URE against 5-HT1A receptor.

3). Li S et al. Fibroblast growth factor 2 contributes to the effect of salidroside on dendritic and synaptic plasticity after cerebral ischemia/reperfusion injury. Aging (Albany NY) 2020 Jun 9;12. (PubMed: 32518214) [IF=5.955]

Application: WB Species: rat Sample:

Figure 6. |Sal upregulates the FGF2-mediated cAMP/PKA/CREB signaling pathway following MCAO/R. (A–G) Representative western blot bands of cAMP, PKA, CREB, p-CREB, FGF2 and FGFR1 in each group.

4). Han X et al. Metformin Modulates High Glucose-Incubated Human Umbilical Vein Endothelial Cells Proliferation and Apoptosis Through AMPK/CREB/BDNF Pathway. Front Pharmacol 2018 Nov 6;9:1266 (PubMed: 30459620) [IF=5.810]

Application: WB Species: human Sample: HUVECs

FIGURE 5 | BDNF expression was regulated by metformin through CREB activation. (A) Western blot analysis of CREB and pCREB expression in HUVECs treated with NG, HG (33.3 mmol/l), and HG + MET (0.01 mmol/l) conditions. Histograms represent the relative pCREB/CREB intensity.

5). Wang C et al. β-Estradiol antagonizes the inhibitory effects of caffeine in BMMSCs via the ERβ-mediated cAMP-dependent PKA pathway. Toxicology 2018 Feb 1;394:1-10 (PubMed: 29154944) [IF=4.571]

Application: WB Species: rat Sample: BMMSCs

Fig. 6.|β-Estradiol antagonizes the caffeine-induced inhibition of osteogenesis by regulating the cAMP/PKA signaling pathway. The PKA activator IBMX and the PKA inhibitor H-89 were added to the culture media.(E) ALP activity, (F) cAMP, (G) CREB gene expression and (H) CREB protein phosphorylation levels were measured by spectrophotometry, ELISA, qRT-PCR, and western blot, respectively. Comparisons between groups were performed using ANOVA,n = 5; #P < 0.05, significantly different from the control group; *P < 0.05, significant difference between each group. ca: caffeine, es: β-estradiol.

6). Chen S et al. The Role of cAMP-PKA Pathway in Lactate-Induced Intramuscular Triglyceride Accumulation and Mitochondria Content Increase in Mice. Front Physiol 2021 Sep 13;12:709135. (PubMed: 34588991) [IF=4.566]

Application: WB Species: Mice Sample: adipose tissue

Figure 3 The time-dependent effects of acute lactate and forskolin injection on the cAMP-PKA pathway. (A) Western blot analysis of cAMP-PKA pathway proteins after acute forskolin injection. (B) Western blot analysis of cAMP-PKA pathway proteins after acute lactate injection. (C,D) The ratios of P-PKA/PKA and P-CREB/CREB after acute forskolin injection. (E,F) The ratios of P-PKA/PKA and P-CREB/CREB after acute lactate injection. AL-15, AL-30, AL-60, and AL-120 represent 15, 30, 60, and 120min after acute lactate injections. AF-15, AF-30, AF-60, and AF-120 represent 15, 30, 60, and 120min after acute forskolin injections. AP and AD mean sacrificed after acute PBS and DMSO injection, respectively. Three bands are used for statistics. The data are presented as the mean±SD, and significant differences between the two groups were analyzed with the independent-samples t-test. * p<0.05 vs. AD and AP.

7). Wu J et al. Zhile Capsule Exerts Antidepressant-Like Effects through Upregulation of the BDNF Signaling Pathway and Neuroprotection. Int J Mol Sci 2019 Jan 8;20(1) (PubMed: 30625977) [IF=4.556]

Application: WB Species: rat Sample: hippocampus

Figure 6.| ZL activated cAMP-CREB-BDNF signaling. (A–C) Protein expressions of BDNF, p-CREB and CREB in the hippocampus at the end of experiment determined by western blotting.Quantified protein level was normalized to that of GAPDH using densitometry (n = 3)

8). Yu L et al. CREB1 protects against the renal injury in a rat model of kidney stone disease and calcium oxalate monohydrate crystals-induced injury in NRK-52E cells. Toxicol Appl Pharmacol 2021 Jan 7;413:115394. (PubMed: 33421503) [IF=4.460]

Application: IF/ICC Species: rat Sample: renal tissue

Fig. 1. CREB1 was activated in kidney stone disease (KSD) model of rats. Rats were injected with 2 × 107 TU lentivirus (LV)-CREB1 vector to enhance the expression of CREB1 or injected with LV-control vector. After 48 h, KSD model was established by treating the rats with 1% (v/v) ethylene glycol +1% (v/v) ammonium chloride by gavage administration daily for four weeks. (A) Immunofluorescence staining for p-CREB1 in renal tissue of control and KSD rats. Western blot analysis for (B) pPKA/PKA and (C) p-CREB1/CREB1 in renal tissue of the rats. Relative protein expression of (D) PKA, (E) p-PKA, (F) CREB1 and (G) p-CREB1 in renal tissue of the rats. + p < 0.05, ++ p < 0.01 and +++ p < 0.001, compared with Control group. * p < 0.05 and *** p < 0.001, compared with KSD + LV-control group.

Application: WB Species: rat Sample: renal tissue

9). Ou J et al. Sinomenine Protects Against Morphine Dependence through the NMDAR1/CAMKII/CREB Pathway: A Possible Role of Astrocyte-Derived Exosomes. Molecules 2018 Sep 17;23(9) (PubMed: 30227624) [IF=4.411]

Application: WB Species: mouse Sample: SH-SY5Y cells

Figure 6. |Effects of astrocyte-derived exosomes on morphine-treated SH-SY5Y cells and subsequent intervention by sinomenine.(C–E) Effects of Sino-exo on p-NMDAR1/NMDAR1, p-CAMKII/CAMKII, and p-CREB/CREB induced by mor-exo in the morphine-treated SH-SY5Y cells (n = 3)

10). Wu Q et al. Electroacupuncture improves neuronal plasticity through the A2AR/cAMP/PKA signaling pathway in SNL rats. Neurochem Int 2021 May;145:104983. (PubMed: 33577869) [IF=4.297]

Application: WB Species: rat Sample: spinal dorsal horn

Fig. 7. EA upregulated the expression of A2AR, cAMP, PKA and p-CREB in the spinal dorsal horn after SNL, while SCH58261 had the opposite effects. (A), (C), (D), (E) Representative Western blots and quantification data of the expression of A2AR/GAPDH, PKA/GAPDH and p-CREB/CREB in each group. n = 5. (B) The content of cAMP in the spinal dorsal horn measured by ELISA. n = 6. (F), (G) qPCR showing the expression of A2AR and PKA mRNA in the spinal cord. n = 6. (H) Immunofluorescence of A2AR-, cAMP- and PKA-positive cells in the spinal dorsal horn. Scale bars, 200 μm. (I), (J), (K) Number of A2AR-, cAMP- and PKA-positive cells in the spinal dorsal horn (lamina I-II and lamina III-IV). n = 4. Columns represent the mean ± SD. $$p < 0.01 vs. the S group; ##p < 0.01 vs. the M group; &&p < 0.01 vs. the M + EA group.

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Phospho-CREB (Ser133) Antibody - #AF3189