Overview

  • Product name

    Anti-STAT5b antibody [EPR16671]
    See all STAT5b primary antibodies
  • Description

    Rabbit monoclonal [EPR16671] to STAT5b
  • Host species

    Rabbit
  • Specificity

    ab178941 shows no cross reactivity with STAT5a.
  • Tested applications

    Suitable for: WB, IHC-P, ICC/IF, IP, Flow Cytmore details
  • Species reactivity

    Reacts with: Mouse, Rat, Human
  • Immunogen

    Recombinant fragment within Mouse STAT5b aa 600 to the C-terminus. The exact sequence is proprietary.
    Database link: P42232

  • Positive control

    • WB: K562, HeLa, Jurkat, Daudi whole cell lysates. C6, RAW 264.7, PC-12 and NIH/3T3 whole cell lysates. Mouse and Rat brain, heart, kidney and spleen lysates. Human fetal heart, kidney and spleen lysates. IHC-P: Rat colon, Mouse spleen and Human spleen tissues. ICC/IF: HeLa cells. IP: K562 whole cell extract.
  • 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.

    We are constantly working hard to ensure we provide our customers with best in class antibodies. As a result of this work we are pleased to now offer this antibody in purified format. We are in the process of updating our datasheets. The purified format is designated 'PUR' on our product labels. If you have any questions regarding this update, please contact our Scientific Support team.

    This product is a recombinant rabbit monoclonal antibody.

Properties

Applications

Our Abpromise guarantee covers the use of ab178941 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
WB 1/5000. Detects a band of approximately 90 kDa (predicted molecular weight: 90 kDa).
IHC-P 1/500. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
ICC/IF 1/100.
IP 1/40.
Flow Cyt 1/40.

Target

  • Function

    Carries out a dual function: signal transduction and activation of transcription. Mediates cellular responses to the cytokine KITLG/SCF and other growth factors. Binds to the GAS element and activates PRL-induced transcription.
  • Involvement in disease

    Growth hormone insensitivity with immunodeficiency
  • Sequence similarities

    Belongs to the transcription factor STAT family.
    Contains 1 SH2 domain.
  • Post-translational
    modifications

    Tyrosine phosphorylated in response to signaling via activated KIT, resulting in translocation to the nucleus. Tyrosine phosphorylated in response to signaling via activated FLT3; wild-type FLT3 results in much weaker phosphorylation than constitutively activated mutant FLT3. Alternatively, can be phosphorylated by JAK2. Phosphorylation at Tyr-699 by PTK6 or HCK leads to an increase of its transcriptional activity. Dephosphorylation on tyrosine residues by PTPN2 negatively regulates prolactin signaling pathway.
  • Cellular localization

    Cytoplasm. Nucleus. Translocated into the nucleus in response to phosphorylation.
  • Information by UniProt
  • Database links

  • Alternative names

    • Signal transducer and activator of transcription 5B antibody
    • STA5B_HUMAN antibody
    • STAT5 antibody
    • Stat5b antibody
    • Transcription factor STAT5B antibody
    see all

Images

  • All lanes : Anti-STAT5b antibody [EPR16671] (ab178941) at 1/20000 dilution

    Lane 1 : K562 (Human chronic myelogenous leukemia cells from bone marrow) whole cell lysates
    Lane 2 : HeLa (Human epithelial cells from cervix adenocarcinoma) whole cell lysates
    Lane 3 : Jurkat (Human T cell leukemia cells from peripheral blood) whole cell lysates
    Lane 4 : Daudi (Human Burkitt's lymphoma cell line) whole cell lysates

    Lysates/proteins at 20 µg per lane.

    Secondary
    All lanes : Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated at 1/1000 dilution

    Predicted band size: 90 kDa
    Observed band size: 90 kDa



    Blocking/dilution buffer: 5% NFDM/TBST.

  • Immunofluorescent analysis of 4% paraformaldehyde-fixed HeLa (Human epithelial cells from cervix adenocarcinoma) cells labeling STAT5b with ab178941 at 1/100 dilution. The cells were permeabilised with 0.1% Triton X-100. Goat anti-rabbit IgG (Alexa Fluor® 488) (ab150077) at 1/200 dilution was used as the secondary antibody (green). Nuclear and cytoplasm staining is detected. The nuclear counter stain is DAPI (blue). Tubulin is detected with ab7291 (Tubulin mouse mAb) at 1/500 and ab150120 (AlexaFluor®594 Goat anti-Mouse secondary) at 1/400 dilution (red). 

     

    The negative controls are as follows;

    1. ab178941 at 1/100 dilution followed by Goat anti mouse IgG (Alexa Fluor®594) at 1/400 dilution.
    2. ab7291 (anti-Tubulin mouse mAb) at 1/500 dilution followed by Goat anti rabbit IgG (Alexa Fluor®488) ar 1/200 dilution.

     

     

  • Immunohistochemical analysis of paraffin-embedded Human spleen tissue labeling STAT5b with ab178941 at 1/500 dilution, followed by prediluted HRP Polymer for Rabbit/Mouse IgG. Nucleus staining on lymphocytes of Human spleen is detected. The negative control utilised PBS instead of primary antibody. Counter stained with Hematoxylin.

    Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

  • ab178941 staining STAT5b in the human cell line HeLa (human cervix adenocarcinoma) by flow cytometry. Cells were fixed with 4% paraformaldehyde and the sample was incubated with the primary antibody at a dilution of 1/40. A goat anti rabbit IgG (Alexa Fluor® 488) at a dilution of 1/2000 was used as the secondary antibody.

    Isoytype control: Rabbit monoclonal IgG (Black)

    Unlabelled control: Cell without incubation with primary antibody and secondary antibody (Blue)

  • All lanes : Anti-STAT5b antibody [EPR16671] (ab178941) at 1/5000 dilution

    Lane 1 : Mouse brain lysates
    Lane 2 : Mouse heart lysates
    Lane 3 : Mouse kidney lysates
    Lane 4 : Mouse spleen lysates
    Lane 5 : Rat brain lysates
    Lane 6 : Rat heart lysates
    Lane 7 : Rat kidney lysates
    Lane 8 : Rat spleen lysates
    Lane 9 : C6 (Rat glial tumor cells) whole cell lysates
    Lane 10 : RAW 264.7 (Mouse macrophage cells transformed with Abelson murine leukemia virus) whole cell lysates
    Lane 11 : PC-12 (Rat adrenal gland pheochromocytoma) whole cell lysates
    Lane 12 : NIH/3T3 (Mouse embyro fibroblast cells) whole cell lysates

    Lysates/proteins at 10 µg per lane.

    Secondary
    All lanes : Anti-Rabbit IgG (HRP), specific to the non-reduced form of IgG at 1/1000 dilution

    Predicted band size: 90 kDa
    Observed band size: 90 kDa



    Blocking/dilution buffer: 5% NFDM/TBST.

  • All lanes : Anti-STAT5b antibody [EPR16671] (ab178941) at 1/20000 dilution

    Lane 1 : Human fetal heart lysates
    Lane 2 : Human fetal kidney lysates
    Lane 3 : Human fetal spleen lysates

    Lysates/proteins at 20 µg per lane.

    Secondary
    All lanes : Anti-Rabbit IgG (HRP), specific to the non-reduced form of IgG at 1/1000 dilution

    Predicted band size: 90 kDa
    Observed band size: 90 kDa



    Blocking/dilution buffer: 5% NFDM/TBST.

  • Lane 1 : Anti-STAT5b antibody [EPR16671] (ab178941) at 1/1000 dilution
    Lane 2 : Anti-STAT5a antibody [E289] (ab32043) at 1/5000 dilution

    All lanes : STAT5a recombinant protein

    Developed using the ECL technique.

    Predicted band size: 90 kDa



    WB showing no cross reactivity with STAT5a.

  • Immunohistochemical analysis of paraffin-embedded Mouse spleen tissue labeling STAT5b with ab178941 at 1/500 dilution, followed by prediluted HRP Polymer for Rabbit/Mouse IgG. Nucleus staining on lymphocytes of Mouse spleen is detected. The negative control utilised PBS instead of primary antibody. Counter stained with Hematoxylin.

    Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

  • Immunohistochemical analysis of paraffin-embedded Rat colon tissue labeling STAT5b with ab178941 at 1/500 dilution, followed by prediluted HRP Polymer for Rabbit/Mouse IgG. Nucleus staining on lymphocytes and weak nucleus staining on gland epithelium of colon is detected. The negative control utilised PBS insead of primary antibody and the slide is counter stained with Hematoxylin.

    Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

  • Immunoprecipitation of K562 (Human chronic myelogenous leukemia cells from bone marrow) whole cell extract using ab178941 at 1/40 dilution. Western blot detection of STAT5b utilised ab178941 at 1/2000 dilution and Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated secondary antibody at 1/1000 dilution. The blocking and dilution buffer was 5% NFDM/TBST.

  • Cross Immunoprecipitation of K562 (Human chronic myelogenous leukemia cells from bone marrow) whole cell extract showing no cross reactivity with STAT5a. Protein captured by anti-STAT5a antibody (ab32042) was detected by the same antibody in WB (image 1) but not by anti-STAT5b, ab178941 (image 2).

     

    For WB detection, ab178941 was used at a 1/2000 dilution and Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated secondary antibody at a 1/1000 dilution. The blocking and dilution buffer was 5% NFDM/TBST.

  • Flow cytometry analysis of 2% paraformaldehyde K562 (Human chronic myelogenous leukemia cells from bone marrow) cells labeling STAT5b with ab178941 at 1:60 dilution (red line). Secondary antibody used is a goat anti rabbit IgG (FITC) at 1:150 dilution. The isotype control is rabbit monoclonal IgG (black line).

References

This product has been referenced in:

See all 4 Publications for this product

Customer reviews and Q&As

1-3 of 3 Abreviews or Q&A

Application
Western blot
Sample
Human Cell lysate - whole cell (prostate cancer, DU-145)
Gel Running Conditions
Reduced Denaturing (10% gel)
Loading amount
20 µg
Specification
prostate cancer, DU-145
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: RT°C

Abcam user community

Verified customer

Submitted Mar 14 2019

Application
Western blot
Sample
Human Cell lysate - whole cell (monocyte)
Gel Running Conditions
Reduced Denaturing (10%)
Loading amount
10 µg
Specification
monocyte
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C

Matija Hedl

Verified customer

Submitted Mar 02 2016

Application
Western blot
Loading amount
15 µg
Gel Running Conditions
Reduced Denaturing (10%)
Sample
Mouse Cell lysate - whole cell (dendritic cells)
Specification
dendritic cells
Treatment
50 ng/ml GM-CSF (6hrs) or LPS 1´g/ml(24hrs)
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: RT°C

Dr. A Amrani

Verified customer

Submitted Oct 27 2014

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