Product: JAK2 Antibody
Catalog: AF6022
Source: Rabbit
Application: WB, IHC, IF/ICC, ELISA(peptide)
Reactivity: Human, Mouse, Rat
Prediction: Pig, Bovine, Horse, Sheep, Rabbit, Dog, Chicken, Xenopus
Mol.Wt.: 125kD; 131kD(Calculated).
Uniprot: O60674
RRID: AB_2834956

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Product Info

Source:
Rabbit
Application:
WB 1:500-1:2000, IHC 1:50-1:200, IF/ICC 1:100-1:500, ELISA(peptide) 1:20000-1:40000
*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:
JAK2 Antibody detects endogenous levels of total JAK2.
RRID:
AB_2834956
Cite Format: Affinity Biosciences Cat# AF6022, RRID:AB_2834956.
Purification:
The antiserum was purified by peptide affinity chromatography using SulfoLink™ Coupling Resin (Thermo Fisher Scientific).
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

JAK 2; JAK-2; JAK2; JAK2_HUMAN; Janus Activating Kinase 2; Janus kinase 2 (a protein tyrosine kinase); Janus kinase 2; JTK 10; JTK10; kinase Jak2; OTTHUMP00000043260; THCYT3; Tyrosine protein kinase JAK2; Tyrosine-protein kinase JAK2;

Immunogens

Immunogen:
Uniprot:
Gene(ID):
Expression:
O60674 JAK2_HUMAN:

Ubiquitously expressed throughout most tissues.

Description:
This gene product is a protein tyrosine kinase involved in a specific subset of cytokine receptor signaling pathways. It has been found to be constituitively associated with the prolactin receptor and is required for responses to gamma interferon.
Sequence:
MGMACLTMTEMEGTSTSSIYQNGDISGNANSMKQIDPVLQVYLYHSLGKSEADYLTFPSGEYVAEEICIAASKACGITPVYHNMFALMSETERIWYPPNHVFHIDESTRHNVLYRIRFYFPRWYCSGSNRAYRHGISRGAEAPLLDDFVMSYLFAQWRHDFVHGWIKVPVTHETQEECLGMAVLDMMRIAKENDQTPLAIYNSISYKTFLPKCIRAKIQDYHILTRKRIRYRFRRFIQQFSQCKATARNLKLKYLINLETLQSAFYTEKFEVKEPGSGPSGEEIFATIIITGNGGIQWSRGKHKESETLTEQDLQLYCDFPNIIDVSIKQANQEGSNESRVVTIHKQDGKNLEIELSSLREALSFVSLIDGYYRLTADAHHYLCKEVAPPAVLENIQSNCHGPISMDFAISKLKKAGNQTGLYVLRCSPKDFNKYFLTFAVERENVIEYKHCLITKNENEEYNLSGTKKNFSSLKDLLNCYQMETVRSDNIIFQFTKCCPPKPKDKSNLLVFRTNGVSDVPTSPTLQRPTHMNQMVFHKIRNEDLIFNESLGQGTFTKIFKGVRREVGDYGQLHETEVLLKVLDKAHRNYSESFFEAASMMSKLSHKHLVLNYGVCVCGDENILVQEFVKFGSLDTYLKKNKNCINILWKLEVAKQLAWAMHFLEENTLIHGNVCAKNILLIREEDRKTGNPPFIKLSDPGISITVLPKDILQERIPWVPPECIENPKNLNLATDKWSFGTTLWEICSGGDKPLSALDSQRKLQFYEDRHQLPAPKWAELANLINNCMDYEPDFRPSFRAIIRDLNSLFTPDYELLTENDMLPNMRIGALGFSGAFEDRDPTQFEERHLKFLQQLGKGNFGSVEMCRYDPLQDNTGEVVAVKKLQHSTEEHLRDFEREIEILKSLQHDNIVKYKGVCYSAGRRNLKLIMEYLPYGSLRDYLQKHKERIDHIKLLQYTSQICKGMEYLGTKRYIHRDLATRNILVENENRVKIGDFGLTKVLPQDKEYYKVKEPGESPIFWYAPESLTESKFSVASDVWSFGVVLYELFTYIEKSKSPPAEFMRMIGNDKQGQMIVFHLIELLKNNGRLPRPDGCPDEIYMIMTECWNNNVNQRPSFRDLALRVDQIRDNMAG

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
0
Model Confidence:
High(score>80) Medium(80>score>50) Low(score<50) No confidence

PTMs - O60674 As Substrate

Site PTM Type Enzyme
Y119 Phosphorylation O60674 (JAK2)
K167 Sumoylation
T174 Phosphorylation
Y201 Phosphorylation O60674 (JAK2)
Y206 Phosphorylation O60674 (JAK2)
K207 Ubiquitination
K212 Ubiquitination
Y221 Phosphorylation O60674 (JAK2)
K244 Ubiquitination
K269 Ubiquitination
Y317 Phosphorylation O60674 (JAK2)
S358 Phosphorylation
Y372 Phosphorylation
Y373 Phosphorylation
Y382 Phosphorylation
Y423 Phosphorylation
Y435 Phosphorylation
S473 Phosphorylation
S518 Phosphorylation
T522 Phosphorylation
S523 Phosphorylation P27361 (MAPK3) , O60674 (JAK2)
Y570 Phosphorylation O60674 (JAK2)
K581 Ubiquitination
K630 Sumoylation
Y637 Phosphorylation O60674 (JAK2)
K728 Ubiquitination
K762 Ubiquitination
Y790 Phosphorylation
Y813 Phosphorylation O60674 (JAK2)
Y868 Phosphorylation O60674 (JAK2)
K882 Ubiquitination
K883 Ubiquitination
K903 Ubiquitination
S904 Phosphorylation
K912 Sumoylation
K914 Sumoylation
Y931 Phosphorylation
Y934 Phosphorylation
S936 Phosphorylation
Y940 Phosphorylation
Y956 Phosphorylation
Y966 Phosphorylation O60674 (JAK2)
Y972 Phosphorylation O60674 (JAK2)
K991 Sumoylation
K999 Ubiquitination
Y1007 Phosphorylation P40189 (IL6ST) , P00519 (ABL1) , O60674 (JAK2)
Y1008 Phosphorylation O60674 (JAK2) , P40189 (IL6ST)
K1011 Sumoylation
K1011 Ubiquitination
K1069 Acetylation

PTMs - O60674 As Enzyme

Substrate Site Source
O14543 (SOCS3) Y204 Uniprot
O14543 (SOCS3) Y221 Uniprot
O14744 (PRMT5) Y297 Uniprot
O14744 (PRMT5) Y304 Uniprot
O14744 (PRMT5) Y307 Uniprot
O60674 (JAK2) Y119 Uniprot
O60674 (JAK2) Y201 Uniprot
O60674 (JAK2) Y206 Uniprot
O60674 (JAK2) Y221 Uniprot
O60674 (JAK2) Y317 Uniprot
O60674 (JAK2) S523 Uniprot
O60674 (JAK2) Y570 Uniprot
O60674 (JAK2) Y637 Uniprot
O60674 (JAK2) Y813 Uniprot
O60674 (JAK2) Y868 Uniprot
O60674 (JAK2) Y966 Uniprot
O60674 (JAK2) Y972 Uniprot
O60674 (JAK2) Y1007 Uniprot
O60674 (JAK2) Y1008 Uniprot
P00533-1 (EGFR) Y1069 Uniprot
P00533 (EGFR) Y1092 Uniprot
P04049-1 (RAF1) Y340 Uniprot
P04049-1 (RAF1) Y341 Uniprot
P10912 (GHR) Y332 Uniprot
P10912 (GHR) Y487 Uniprot
P16410 (CTLA4) Y201 Uniprot
P19235-1 (EPOR) Y368 Uniprot
P19235-1 (EPOR) Y426 Uniprot
P19235-1 (EPOR) Y454 Uniprot
P19235-1 (EPOR) Y456 Uniprot
P19235-1 (EPOR) Y468 Uniprot
P19235-1 (EPOR) Y485 Uniprot
P19235-1 (EPOR) Y489 Uniprot
P19235-1 (EPOR) Y504 Uniprot
P27361 (MAPK3) Y204 Uniprot
P28482 (MAPK1) Y187 Uniprot
P40763-2 (STAT3) Y704 Uniprot
P40763-3 (STAT3) Y705 Uniprot
P41597 (CCR2) Y139 Uniprot
P42224 (STAT1) Y701 Uniprot
P42229-1 (STAT5A) Y694 Uniprot
P42680 (TEC) Y519 Uniprot
P46527 (CDKN1B) Y88 Uniprot
P51692 (STAT5B) Y699 Uniprot
P62993 (GRB2) Y7 Uniprot
P62993 (GRB2) Y37 Uniprot
P62993 (GRB2) Y52 Uniprot
P62993 (GRB2) Y209 Uniprot
P78324-1 (SIRPA) Y496 Uniprot
P78347 (GTF2I) Y248 Uniprot
P84243 (H3F3B) Y42 Uniprot
Q13153 (PAK1) Y153 Uniprot
Q13153 (PAK1) Y201 Uniprot
Q13153 (PAK1) Y285 Uniprot
Q14765 (STAT4) Y693 Uniprot
Q15118 (PDK1) Y243 Uniprot
Q15910 (EZH2) Y641 Uniprot
Q92888 (ARHGEF1) Y738 Uniprot
Q99683 (MAP3K5) Y718 Uniprot
Q9UGK3 (STAP2) Y22 Uniprot
Q9UGK3-2 (STAP2) Y250 Uniprot
Q9UGK3 (STAP2) Y310 Uniprot
Q9UGK3 (STAP2) Y322 Uniprot
Q9UQC2 (GAB2) Y643 Uniprot

Research Backgrounds

Function:

Non-receptor tyrosine kinase involved in various processes such as cell growth, development, differentiation or histone modifications. Mediates essential signaling events in both innate and adaptive immunity. In the cytoplasm, plays a pivotal role in signal transduction via its association with type I receptors such as growth hormone (GHR), prolactin (PRLR), leptin (LEPR), erythropoietin (EPOR), thrombopoietin (THPO); or type II receptors including IFN-alpha, IFN-beta, IFN-gamma and multiple interleukins. Following ligand-binding to cell surface receptors, phosphorylates specific tyrosine residues on the cytoplasmic tails of the receptor, creating docking sites for STATs proteins. Subsequently, phosphorylates the STATs proteins once they are recruited to the receptor. Phosphorylated STATs then form homodimer or heterodimers and translocate to the nucleus to activate gene transcription. For example, cell stimulation with erythropoietin (EPO) during erythropoiesis leads to JAK2 autophosphorylation, activation, and its association with erythropoietin receptor (EPOR) that becomes phosphorylated in its cytoplasmic domain. Then, STAT5 (STAT5A or STAT5B) is recruited, phosphorylated and activated by JAK2. Once activated, dimerized STAT5 translocates into the nucleus and promotes the transcription of several essential genes involved in the modulation of erythropoiesis. Part of a signaling cascade that is activated by increased cellular retinol and that leads to the activation of STAT5 (STAT5A or STAT5B). In addition, JAK2 mediates angiotensin-2-induced ARHGEF1 phosphorylation. Plays a role in cell cycle by phosphorylating CDKN1B. Cooperates with TEC through reciprocal phosphorylation to mediate cytokine-driven activation of FOS transcription. In the nucleus, plays a key role in chromatin by specifically mediating phosphorylation of 'Tyr-41' of histone H3 (H3Y41ph), a specific tag that promotes exclusion of CBX5 (HP1 alpha) from chromatin.

PTMs:

Autophosphorylated, leading to regulate its activity. Leptin promotes phosphorylation on tyrosine residues, including phosphorylation on Tyr-813 (By similarity). Autophosphorylation on Tyr-119 in response to EPO down-regulates its kinase activity (By similarity). Autophosphorylation on Tyr-868, Tyr-966 and Tyr-972 in response to growth hormone (GH) are required for maximal kinase activity (By similarity). Also phosphorylated by TEC (By similarity). Phosphorylated on tyrosine residues in response to interferon gamma signaling. Phosphorylated on tyrosine residues in response to a signaling cascade that is activated by increased cellular retinol.

Subcellular Location:

Endomembrane system>Peripheral membrane protein. Cytoplasm. Nucleus.

Extracellular region or secreted Cytosol Plasma membrane Cytoskeleton Lysosome Endosome Peroxisome ER Golgi apparatus Nucleus Mitochondrion Manual annotation Automatic computational assertionSubcellular location
Tissue Specificity:

Ubiquitously expressed throughout most tissues.

Subunit Structure:

Interacts with EPOR, LYN, SIRPA, SH2B1 and TEC (By similarity). Interacts with IL23R. Interacts with SKB1. Interacts with STAM2. Interacts with IFNGR2 (via intracellular domain). Interacts with LEPR (Isoform B) (By similarity). Interacts with HSP90AB1; promotes functional activation in a heat shock-dependent manner. Interacts with STRA6. Interacts with RHEX; this interaction occurs in a erythropoietin (EPO)-dependent manner.

Family&Domains:

Possesses 2 protein kinase domains. The second one probably contains the catalytic domain, while the presence of slight differences suggest a different role for protein kinase 1 (By similarity).

Belongs to the protein kinase superfamily. Tyr protein kinase family. JAK subfamily.

Research Fields

· Cellular Processes > Cell growth and death > Necroptosis.   (View pathway)

· Cellular Processes > Cellular community - eukaryotes > Signaling pathways regulating pluripotency of stem cells.   (View pathway)

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

· Environmental Information Processing > Signal transduction > Jak-STAT signaling pathway.   (View pathway)

· Human Diseases > Drug resistance: Antineoplastic > EGFR tyrosine kinase inhibitor resistance.

· Human Diseases > Infectious diseases: Parasitic > Leishmaniasis.

· Human Diseases > Infectious diseases: Parasitic > Toxoplasmosis.

· Human Diseases > Infectious diseases: Bacterial > Tuberculosis.

· Human Diseases > Infectious diseases: Viral > Measles.

· Human Diseases > Infectious diseases: Viral > Influenza A.

· Human Diseases > Infectious diseases: Viral > Herpes simplex infection.

· Human Diseases > Cancers: Overview > Pathways in cancer.   (View pathway)

· Organismal Systems > Immune system > Chemokine signaling pathway.   (View pathway)

· Organismal Systems > Immune system > Th1 and Th2 cell differentiation.   (View pathway)

· Organismal Systems > Immune system > Th17 cell differentiation.   (View pathway)

· Organismal Systems > Nervous system > Cholinergic synapse.

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

· Organismal Systems > Endocrine system > Adipocytokine signaling pathway.

References

1). Cao P et al. MiR-15b is a key regulator of proliferation and apoptosis of chondrocytes from patients with condylar hyperplasia by targeting IGF1, IGF1R and BCL2. Osteoarthritis Cartilage 2019 Feb;27(2):336-346 (PubMed: 30521861) [IF=7.507]

2). Fan X et al. Design, synthesis and biological evaluation of N-anthraniloyl tryptamine derivatives as pleiotropic molecules for the therapy of malignant glioma. Eur J Med Chem 2021 Oct 15;222:113564. (PubMed: 34091208) [IF=7.088]

Application: WB    Species: Human    Sample: U251 and U87MG cells

Fig. 2. NP16 down-regulated the expression of COX-2, p-JAK2 and p-STAT3, and inhibited the nuclear translocation of STAT3 in both U251 and U87MG cells. (A, B) western blot analysis of the expression of JAK2, p-JAK2, p-STAT3 and STAT3: the cells were exposed to NP16 at concentrations of 0.25, 1 and 4 mM for 8 h; western blot analysis of the expression of COX-2: the cells were exposed to NP16 at concentrations of 1, 2 and 4 mM for 24 h (C, D, E) Relative expression of the target protein in the cell lysate was calculated using the ImageJ image analysis software. Each error bar represents the mean ± SD of three independent experiments. *P < 0.05, **P < 0.01 and ***P < 0.001 versus the DMSO group. (F, G) representative fluorescence microscopy images: the cells were exposed to NP16 at concentrations of 4 mM for 8 h. Compound NP16 inhibited nuclear translocation of STAT3 in both U251 and U87MG cells. (Magnification  60, scale bar ¼ 20 mm).

3). Wei X et al. Higenamine alleviates allergic rhinitis by activating AKT1 and suppressing the EGFR/JAK2/c-JUN signaling. Phytomedicine 2021 Jun;86:153565. (PubMed: 33945919) [IF=6.656]

Application: WB    Species: Human    Sample: HNEpCs

Fig. 8. The effects of higenamine (HG) on potential targets in histamine-induced HNEpCs. (A-J) Levels of protein expression of AKT1, p-AKT1, EGFR, p-EGFR, c-Jun, p-c-Jun, iNOS, JAK2, and p-JAK2 were determined by Western blotting, β-actin was used as an internal control. Data are expressed as means ± SD of three experiments. ## p < 0.01, and # p < 0.05 compared with the control group. ** p < 0.01, and * p < 0.05 compared with the histamine group.

4). Liu N et al. Hyperuricemia induces lipid disturbances mediated by LPCAT3 upregulation in the liver. FASEB J 2020 Aug 11. (PubMed: 32780898) [IF=5.834]

Application: WB    Species: human    Sample: L02 cells

FIGURE 6 |Uric acid induces SREBP-1c activation and JAK2/STAT3 pathway inhibition in vitro.F, p-JAK2 (Tyr1007/1008)/JAK2 and p-STAT3 (Tyr705)/STAT3 protein levels were detected by western blotting.

5). Zhang Y et al. The Food Additive β-Caryophyllene Exerts Its Neuroprotective Effects Through the JAK2-STAT3-BACE1 Pathway. Front Aging Neurosci 2022 Feb 28;14:814432. (PubMed: 35296033) [IF=5.702]

Application: WB    Species: Rat    Sample: PC-12 cells

FIGURE 5 Effect of β-caryophyllene on JAK2 and STAT3 phosphorylation and BACE1 protein expression in PC-12 cells. CN denotes the control group without special treatment; EVG denotes the empty vector group in which the PC-12 cells were transfected with plasmid vectors without gene fragments; OE denotes the group in which the PC-12 cells were transfected with a plasmid containing APP cDNA; and BCP denotes the group in which the effect of BCP on the 24-h viability of PC-12 cells transfected for 48 h was assessed. (A) Phosphorylation level of JAK2 protein. (B) Phosphorylation level of STAT3 protein. (C) Relative expression level of BACE1 protein. Compared with CN, ns p > 0.05. Compared with EVG, *p < 0.05, **p < 0.01. Compared with OE group, Δp < 0.05, ΔΔp < 0.01. One-way ANOVA with Bonferroni’s post hoc test.

6). Li Z et al. Total flavonoids of Sophora flavescens and kurarinone ameliorated ulcerative colitis by regulating Th17/Treg cell homeostasis. J Ethnopharmacol 2022 Oct 28;297:115500. (PubMed: 35863614) [IF=5.195]

7). Lu Z et al. Huanglian Jiedu Decoction ameliorates DSS-induced colitis in mice via the JAK2/STAT3 signalling pathway. Chin Med 2020 May 8;15:45. (PubMed: 32411291) [IF=4.546]

Application: WB    Species: Mice    Sample: colonic tissue

Fig. 4 Effect of HJD on the protein expression of JAK2 and STAT3 in colon. a Western blotting of JAK2 and STAT3 protein. b Semi-quantitative analysis of JAK2 and STAT3 proteins. c, d Immunofluorescence staining of JAK2 and STAT3 co-localized in control group, UC group, AG490 group, AG490 + HJD group and HJD group. All data are expressed as mean ± SD, **P < 0.01 compared with UC group, and ##P < 0.01 compared with control group. Scale bar = 50 μm

Application: IF/ICC    Species: Mice    Sample: colonic tissue

Fig. 4 Effect of HJD on the protein expression of JAK2 and STAT3 in colon. a Western blotting of JAK2 and STAT3 protein. b Semi-quantitative analysis of JAK2 and STAT3 proteins. c, d Immunofluorescence staining of JAK2 and STAT3 co-localized in control group, UC group, AG490 group, AG490 + HJD group and HJD group. All data are expressed as mean ± SD, **P < 0.01 compared with UC group, and ##P < 0.01 compared with control group. Scale bar = 50 μm

8). Liu J et al. Sphk1 promotes ulcerative colitis via activating JAK2/STAT3 signaling pathway. Hum Cell 2019 Oct 12 (PubMed: 31606874) [IF=4.374]

Application: WB    Species: mouse    Sample: RAW264.7 cells

Fig. 5 |Overexpression of SphK1 signifcantly activated NF-kB and JAK2/STAT3 signaling pathway. a Expression levels of SphK1, SphK2, S1PR1, p-p65, p65, p-STAT3, STAT3, p-jak2 and jak2 in RAW264.7 cells were detected by Western blot. GAPDH was used as an internal control.

9). Chen L et al. Combination of gemcitabine and erlotinib inhibits recurrent pancreatic cancer growth in mice via the JAK-STAT pathway. Oncol Rep 2018 Mar;39(3):1081-1089 (PubMed: 29328487) [IF=4.136]

Application: WB    Species: human    Sample: BxPC-3 and PANC‑1 cells

Figure 3.| The gemcitabine-erlotinib (E+G) combination group inhibits the activity of the JAK-STAT pathway, as well as the expression of downstream HIF‑1α, cyclin D1 and p53. (E) Phosphorylation levels of JAK1 (Tyr1022), JAK2 (Tyr221), JAK3(Tyr981) and STAT3 (Tyr701) in BxPC-3 and PANC‑1 cells were analyzed by western blotting, respectively.

10). Zhu L et al. Cerebroprotein hydrolysate‐I protects senescence‐induced by D‐galactose in PC12 cells and mice. Food Sci Nutr 2021 May 17;9(7):3722-3731. (PubMed: 34262731) [IF=3.553]

Application: WB    Species: Rat    Sample: PC12 cells

FIGURE 4 Effects of CH‐I on related protein expressions of PC12 cells induced by D‐gal. (a) The original bands of BDNF, JAK2, p‐JAK2, STAT3, p‐STAT2, TERT and β‐actin. β‐actin was considered as loading controls. (b) The ratio of different proteins to β‐actin was calculated by the band density. *p < .05, **p < .01 vs control group; # p < .05 and ## p < .01 vs D‐gal group

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