Recombinant
RabMAb

Anti-JAK2 (phospho Y1007 + Y1008) antibody [E132] (ab32101)

Abreviews (2)Q&A (6)Specific References (33)

Overview

  • Product name
    Anti-JAK2 (phospho Y1007 + Y1008) antibody [E132]
    See all JAK2 primary antibodies
  • Description
    Rabbit monoclonal [E132] to JAK2 (phospho Y1007 + Y1008)
  • Host species
    Rabbit
  • Specificity
    The antibody is phospho-specific and only detects phosphorylated JAK2 on Tyrosine 1007 and 1008 (pY1007+Y1008). According to our ELISA results, this antibody preferentially recognizes phospho Y1007. Stimulation may be required to allow detection of the phosphorylated protein. Please see images below for recommended treatment conditions and positive controls.
  • Tested applications
    Suitable for: ELISA, WB, IHC-Fr, ICC/IF, Flow Cyt, IHC-P, IP, Dot blotmore details
  • Species reactivity
    Reacts with: Mouse, Rat, Human
  • Immunogen

    Synthetic peptide (the amino acid sequence is considered to be commercially sensitive) within Human JAK2 (phospho Y1007 + Y1008). The exact sequence is proprietary.

  • Positive control
    • WB: Jurkat or K562 cells (treated with Pervanadate). IHC-P: Human lung adenocarcinoma tissue. ICC/IF: Jurkat cells (treated with Pervanadate). Flow Cyt: Jurkat starved of serum for 16 hours then treated with 1mM Pervanadate for 30 minutes.
  • General notes

    Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMab® patents

    This product is a recombinant rabbit monoclonal antibody.

Properties

Applications

Our Abpromise guarantee covers the use of ab32101 in the following tested applications.

The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.

Application Abreviews Notes
ELISA Use at an assay dependent concentration.
WB 1/1000 - 1/10000. Detects a band of approximately 120 kDa (predicted molecular weight: 130 kDa).

The samples may require stimulation (E.g., Jurkat cells treated with pervanadate for 5 min)
IHC-Fr Use at an assay dependent concentration. PubMed: 21420414
ICC/IF 1/1000.
Flow Cyt 1/20.
IHC-P Use at an assay dependent concentration.
IP 1/50.
Dot blot 1/1000.

Target

  • Function
    Non-receptor tyrosine kinase involved in various processes such as cell cycle progression, apoptosis, mitotic recombination, genetic instability and histone modifications. In the cytoplasm, plays a pivotal role in signal transduction via its association with cytokine receptors, which constitutes an initiating step in signaling for many members of the cytokine receptor superfamily including the receptors for growth hormone (GHR), prolactin (PRLR), leptin (LEPR), erythropoietin (EPOR), granulocyte-macrophage colony-stimulating factor (CSF2), thrombopoietin (THPO) and multiple interleukins. Following stimulation with erythropoietin (EPO) during erythropoiesis, it is autophosphorylated and activated, leading to its association with erythropoietin receptor (EPOR) and tyrosine phosphorylation of residues in the EPOR cytoplasmic domain. Also involved in promoting the localization of EPOR to the plasma membrane. Also acts downstream of some G-protein coupled receptors. Plays a role in the control of body weight (By similarity). Mediates angiotensin-2-induced ARHGEF1 phosphorylation. 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.
  • Tissue specificity
    Expressed in blood, bone marrow and lymph node.
  • Involvement in disease
    Note=Chromosomal aberrations involving JAK2 are found in both chronic and acute forms of eosinophilic, lymphoblastic and myeloid leukemia. Translocation t(8;9)(p22;p24) with PCM1 links the protein kinase domain of JAK2 to the major portion of PCM1. Translocation t(9;12)(p24;p13) with ETV6.
    Defects in JAK2 are a cause of susceptibility to Budd-Chiari syndrome (BCS) [MIM:600880]. It is a syndrome caused by obstruction of hepatic venous outflow involving either the hepatic veins or the terminal segment of the inferior vena cava. Obstructions are generally caused by thrombosis and lead to hepatic congestion and ischemic necrosis. Clinical manifestations observed in the majority of patients include hepatomegaly, right upper quadrant pain and abdominal ascites. Budd-Chiari syndrome is associated with a combination of disease states including primary myeloproliferative syndromes and thrombophilia due to factor V Leiden, protein C deficiency and antithrombin III deficiency. Budd-Chiari syndrome is a rare but typical complication in patients with polycythemia vera.
    Defects in JAK2 are a cause of polycythemia vera (PV) [MIM:263300]. A myeloproliferative disorder characterized by abnormal proliferation of all hematopoietic bone marrow elements, erythroid hyperplasia, an absolute increase in total blood volume, but also by myeloid leukocytosis, thrombocytosis and splenomegaly.
    Defects in JAK2 gene may be a cause of essential thrombocythemia (ET) [MIM:187950]. ET is characterized by elevated platelet levels due to sustained proliferation of megakaryocytes, and frequently lead to thrombotic and haemorrhagic complications.
    Defects in JAK2 are a cause of myelofibrosis (MYELOF) [MIM:254450]. Myelofibrosis is a disorder characterized by replacement of the bone marrow by fibrous tissue, occurring in association with a myeloproliferative disorder. Clinical manifestations may include anemia, pallor, splenomegaly, hypermetabolic state, petechiae, ecchymosis, bleeding, lymphadenopathy, hepatomegaly, portal hypertension.
    Defects in JAK2 are a cause of acute myelogenous leukemia (AML) [MIM:601626]. AML is a malignant disease in which hematopoietic precursors are arrested in an early stage of development.
  • Sequence similarities
    Belongs to the protein kinase superfamily. Tyr protein kinase family. JAK subfamily.
    Contains 1 FERM domain.
    Contains 1 protein kinase domain.
    Contains 1 SH2 domain.
  • Domain
    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.
  • Post-translational
    modifications
    Autophosphorylated, leading to regulate its activity. Leptin promotes phosphorylation on tyrosine residues, including phosphorylation on Tyr-813. Autophosphorylation on Tyr-119 in response to EPO down-regulates its kinase activity. Autophosphorylation on Tyr-868, Tyr-966 and Tyr-972 in response to growth hormone (GH) are required for maximal kinase activity.
  • Cellular localization
    Endomembrane system. Nucleus.
  • Information by UniProt
  • Database links
    see all
  • Alternative names
    • JAK 2 antibody
    • JAK-2 antibody
    • JAK2 antibody
    • JAK2_HUMAN antibody
    • Janus Activating Kinase 2 antibody
    • Janus kinase 2 (a protein tyrosine kinase) antibody
    • Janus kinase 2 antibody
    • JTK 10 antibody
    • JTK10 antibody
    • kinase Jak2 antibody
    • OTTHUMP00000043260 antibody
    • THCYT3 antibody
    • Tyrosine protein kinase JAK2 antibody
    • Tyrosine-protein kinase JAK2 antibody
    see all

Images

  • Immunocytochemistry/ Immunofluorescence - Anti-JAK2 (phospho Y1007 + Y1008) antibody [E132] (ab32101)
    Immunocytochemistry/ Immunofluorescence - Anti-JAK2 (phospho Y1007 + Y1008) antibody [E132] (ab32101)

    Immunocytochemistry/Immunofluorescence analysis of Jurkat +/- pervanadate (1mM, 30min) and Jurkat + pervanadate (1mM, 30min) + LP cells labelling JAK2 (phospho Y1007 + Y1008) with ab32101 at a dilution of 1/1000. Cells were fixed with 4% paraformaldehyde and permeabilized with 0.1% tritonX-100. ab150077 (goat anti-rabbit IgG Alexa Fluor® 488) (1/1000) was used as the secondary antibody. The cells were co-stained with ab195889 (Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor® 594)) at a 1/200 dilution. Nuclei counterstained with DAPI (blue).

  • Western blot - Anti-JAK2 (phospho Y1007 + Y1008) antibody [E132] (ab32101)
    Western blot - Anti-JAK2 (phospho Y1007 + Y1008) antibody [E132] (ab32101)
    All lanes : Anti-JAK2 (phospho Y1007 + Y1008) antibody [E132] (ab32101) at 1/5000 dilution

    Lane 1 : Untreated Jurkat cells whole cell lysates
    Lane 2 : Jurkat cells were treated with 50mM Pervanadate for 5 minutes whole cell lysates
    Lane 3 : Jurkat cells were treated with 50mM Pervanadate for 5 minutes whole cell lysates. Then the membrane was incubated with Alkaline phosphatase.

    Lysates/proteins at 10 µg per lane.

    Secondary
    All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/20000 dilution

    Predicted band size: 130 kDa
    Observed band size: 120 kDa (why is the actual band size different from the predicted?)


    Exposure time: 5 seconds


    Blocking and diluting buffer 5% NFDM/TBST

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-JAK2 (phospho Y1007 + Y1008) antibody [E132] (ab32101)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-JAK2 (phospho Y1007 + Y1008) antibody [E132] (ab32101)
    ab32101 showing positive staining in Lung adenocarcinoma tissue.

  • Dot Blot - Anti-JAK2 (phospho Y1007 + Y1008) antibody [E132] (ab32101)
    Dot Blot - Anti-JAK2 (phospho Y1007 + Y1008) antibody [E132] (ab32101)

    Dot blot analysis of JAK2 (phospho Y1007 & Y1008) phospho peptide (Lane 1), JAK2 (phospho Y1007) phospho peptide (Lane 2), JAK2 (phospho Y1008) phospho peptide (Lane 3) and JAK2 non-phospho peptide (Lane 4) labelling JAK2 (phospho Y1007 & Y1008) with ab32101 at a dilution of 1/1000. A Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated (ab97051) was used as the secondary antibody at a dilution of 1/20,000. Blocking buffer: 5% NFDM/TBST. Dilution buffer: 5% NFDM /TBST.

  • Western blot - Anti-JAK2 (phospho Y1007 + Y1008) antibody [E132] (ab32101)
    Western blot - Anti-JAK2 (phospho Y1007 + Y1008) antibody [E132] (ab32101)
    All lanes : Anti-JAK2 (phospho Y1007 + Y1008) antibody [E132] (ab32101) at 1/1000 dilution

    Lane 1 : Jurkat (Human T cell leukemia T lymphocyte) whole cell lysates
    Lane 2 : Jurkat (Human T cell leukemia T lymphocyte) treated with 50mM Pervanadate for 5 minutes whole cell lysates

    Lysates/proteins at 15 µg per lane.

    Secondary
    All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/20000 dilution

    Predicted band size: 130 kDa
    Observed band size: 120 kDa (why is the actual band size different from the predicted?)
    Additional bands at: 60 kDa. We are unsure as to the identity of these extra bands.



    Blocking and diluting buffer: 5% NFDM/TBST

    Exposure time: 
    Left image: 1 second
    Right image: 5 minutes

  • Flow Cytometry - Anti-JAK2 (phospho Y1007 + Y1008) antibody [E132] (ab32101)
    Flow Cytometry - Anti-JAK2 (phospho Y1007 + Y1008) antibody [E132] (ab32101)

    Flow Cytometry analysis of Jurkat (human acute T cell leukemia) cells starved of serum for 16 hours then treated with 1 mM Pervanadate for 30 minutes labeling JAK2 (phospho Y1007 + Y1008) with ab32101 at 1/20 dilution (10 ug/mL) (red). Cells were fixed with 4% paraformaldehyde and permeabilised with 90% methanol. A Goat anti rabbit IgG (Alexa Fluor®488) at 1/2000 dilution was used as the secondary antibody. Rabbit monoclonal IgG (Black) was used as the isotype control, cells without incubation with primary antibody and secondary antibody (Blue) was used as the unlabeled control. 

  • ELISA - Anti-JAK2 (phospho Y1007 + Y1008) antibody [E132] (ab32101)
    ELISA - Anti-JAK2 (phospho Y1007 + Y1008) antibody [E132] (ab32101)

    Direct ELISA antigen dose-response curve using ab32101 at 0~1000 ng/mL. Antigen concentration of 100 ng/mL. An Alkaline Phosphatase-conjugated AffiniPure Goat Anti-Rabbit IgG(H+L) (1/2500) was used as the secondary antibody.

    This antibody preferentially recognizes phospho Y1007. When the concentration of peptides is higher than 100 ng/mL, it also recognizes phospho Y1008.

  • ELISA - Anti-JAK2 (phospho Y1007 + Y1008) antibody [E132] (ab32101)
    ELISA - Anti-JAK2 (phospho Y1007 + Y1008) antibody [E132] (ab32101)

    Direct ELISA antigen dose-response curve using ab32101 at 0~1000 ng/mL. Antigen concentration of 10 ng/mL. An Alkaline Phosphatase-conjugated AffiniPure Goat Anti-Rabbit IgG(H+L) (1/2500) was used as the secondary antibody.

    This antibody preferentially recognizes phospho Y1007. When the concentration of peptides is lower than 10 ng/mL, it cannot recognize phospho Y1008.

     

  • Dot Blot - Anti-JAK2 (phospho Y1007 + Y1008) antibody [E132] (ab32101)
    Dot Blot - Anti-JAK2 (phospho Y1007 + Y1008) antibody [E132] (ab32101)
    Analysis of immunogen phosphopeptide (A) and non phosphopeptide (B). Antibody used at 1/1000 dilution.
  • Western blot - Anti-JAK2 (phospho Y1007 + Y1008) antibody [E132] (ab32101)
    Western blot - Anti-JAK2 (phospho Y1007 + Y1008) antibody [E132] (ab32101)This image is courtesy of an anonymous Abreview
    All lanes : Anti-JAK2 (phospho Y1007 + Y1008) antibody [E132] (ab32101) at 1/2000 dilution

    Lane 1 : Rat uterine cell line - whole cell lysate. Treated with 1µg/mL Prolactin for 0 hours.
    Lane 2 : Rat uterine cell line - whole cell lysate. Treated with 1µg/mL Prolactin for 15 minutes.
    Lane 3 : Rat uterine cell line - whole cell lysate. Treated with 1µg/mL Prolactin for 30 minutes.
    Lane 4 : Rat uterine cell line - whole cell lysate. Treated with 1µg/mL Prolactin for 1 hour.
    Lane 5 : Rat uterine cell line - whole cell lysate. Treated with 1µg/mL Prolactin for 2 hours.
    Lane 6 : Rat uterine cell line - whole cell lysate. Treated with 1µg/mL Prolactin for 4 hours.
    Lane 7 : Rat uterine cell line - whole cell lysate. Treated with 1µg/mL Prolactin for 6 hours.
    Lane 8 : Rat uterine cell line - whole cell lysate. Treated with 1µg/mL Prolactin for 24 hours.

    Lysates/proteins at 30 µg per lane.

    Secondary
    All lanes : An HRP-conjugated donkey anti-rabbit polyclonal. at 1/10000 dilution

    Developed using the ECL technique.

    Performed under reducing conditions.

    Predicted band size: 130 kDa
    Observed band size: 110 kDa (why is the actual band size different from the predicted?)
    Additional bands at: 55 kDa (possible non-specific binding)

    See Abreview

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-JAK2 (phospho Y1007 + Y1008) antibody [E132] (ab32101)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-JAK2 (phospho Y1007 + Y1008) antibody [E132] (ab32101)
    ab32101 showing positive staining in Ovarian carcinoma tissue.

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-JAK2 (phospho Y1007 + Y1008) antibody [E132] (ab32101)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-JAK2 (phospho Y1007 + Y1008) antibody [E132] (ab32101)
    ab32101 showing positive staining in Cervical carcinoma tissue.

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-JAK2 (phospho Y1007 + Y1008) antibody [E132] (ab32101)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-JAK2 (phospho Y1007 + Y1008) antibody [E132] (ab32101)
    ab32101 showing positive staining in Endometrial carcinoma tissue.

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-JAK2 (phospho Y1007 + Y1008) antibody [E132] (ab32101)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-JAK2 (phospho Y1007 + Y1008) antibody [E132] (ab32101)
    ab32101 showing positive staining in Urinary bladder transitional carcinoma tissue.

References

This product has been referenced in:

See all 33 Publications for this product

Publishing research using ab32101? Please let us know so that we can cite the reference in this datasheet.

Customer reviews and Q&As

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8 reviews or Q&A

10
Votes: Highest First

I purchased 12 antibodies from your company.
Some didn't work and I need your help.

I used mouse liver tissue. I attached our lab's protocol.
Apply 25ug of protein in each well.
The gel I used is NuPAGE 4-12% Bis-Tris Gel.

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Thank you for bringing this to our attention. I am sorry to read that two of the antibodies you purchased are not staining blots of your mouse liver samples.

I am assuming the transfer from gel to blot was effective, if your protocol worked ...

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What is the conc of lot GR18831-6 (ab32101)

Thank you for contacting us.

The concentration of lot GR57259-1 is XXXXXXX mg/ml.

I hope this information is helpful to you. Please do not hesitate to contact us if you need any more advice or information.

Use our products? S...

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I am very interested in purchasing JAK2 (phospho Y1007 + Y1008)
antibody [E132] (ab32101) , but I would know whic positive control
(name of cell line, type and time of treatment) is used in western
blot image and moreover if this lysate ...

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Thank you for your enquiry.

I can confirm that this antibody has been tested on the following and these would be suitable positive controls:

Rat uterine cell line Treated with 1µg/mL Prolactin for 1 hour.

Jurkat c...

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Haben Sie Erfahrungen zur Verwendung dieses AKs an Gefrierschnitten?

Vielen Dank für Ihren Anruf und für Ihr Interesse an unseren Produkten. Gerne versichere ich Ihnen, dass alle unsere Produkte unsere Abcam Garantier unterliegen: Falls sich herausstellt, dass der Antikörper nicht so funktioniert, wie auf dem Datenbl...

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I have a question regarding the treatment of tissue prior to applying the primary antibody. Do you recommend microwave? Enzymatic? The tissue I have in mind is human salivary gland. I'm planning to use ab6672, ab59389, ab39636, ab32101, ab31...

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Thank you for contacting us. The treatment of PFA-fixed, paraffin embedded sections (IHC-P) before incubation with priamry antibody would include antigen retrieval, blocking of endogenous peroxidases (if HRP is being used for detection), and bl...

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Western blot abreview for Anti-JAK2 (phospho Y1007 + Y1008) antibody [E132]

 Excellent
Abreviews
Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Western blot
Sample
Rat Cell lysate - whole cell (Uterine Cell line)
Loading amount
30 µg
Specification
Uterine Cell line
Treatment
1ug/ml Prolactin
Gel Running Conditions
Reduced Denaturing (10)
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 24°C
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Submitted Mar 19 2010

Western blot abreview for Anti-JAK2 (phospho Y1007 + Y1008) antibody [E132]

 Poor
Abreviews
Application
Western blot
Sample
Human Cell lysate - whole cell (Lung)
Loading amount
10 µg
Specification
Lung
Treatment
10ng/mL IFN gamma for 24hrs
Gel Running Conditions
Reduced Denaturing
Blocking step
Milk as blocking agent for 18 hour(s) and 0 minute(s) · Concentration: 10% · Temperature: 4°C
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Username

Abcam user community

Verified customer

Submitted Jul 31 2007

Dear Sirs, I am potentially very interested in purchasing this mAb but I would like first to know: 1) whether this mAb is specific for the Y1007-Y1008 phosphorylated form of JAK2 or also detects the non-phosphorylated form (could you please provide a ...

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Thank you for contacting us. In answer to your questions: 1. The antibody ab32101 is specific to the phosphorylated protein. We do not have another Western blot but we are confident, given the dot blot result, that this antibody will not recognize ...

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