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  1. Link

    shp1-antibody-epr5519-ab124942.pdf

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Cell Biology Cell Cycle Kinases/Phosphatases Phosphatases
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Validated using a knockout cell lineRecombinantRabMAb

Recombinant Anti-SHP1 antibody [EPR5519] (ab124942)

  • Datasheet
  • SDS
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Western blot - Anti-SHP1 antibody [EPR5519] (ab124942)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-SHP1 antibody [EPR5519] (ab124942)
  • Immunocytochemistry/ Immunofluorescence - Anti-SHP1 antibody [EPR5519] (ab124942)
  • Western blot - Anti-SHP1 antibody [EPR5519] (ab124942)
  • Flow Cytometry (Intracellular) - Anti-SHP1 antibody [EPR5519] (ab124942)
  • OI-RD Scanning - Anti-SHP1 antibody [EPR5519] (ab124942)
  • Anti-SHP1 antibody [EPR5519] (ab124942)

Key features and details

  • Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
  • Rabbit monoclonal [EPR5519] to SHP1
  • Suitable for: Flow Cyt (Intra), WB, IHC-P, ICC/IF
  • Knockout validated
  • Reacts with: Human

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Overview

  • Product name

    Anti-SHP1 antibody [EPR5519]
    See all SHP1 primary antibodies
  • Description

    Rabbit monoclonal [EPR5519] to SHP1
  • Host species

    Rabbit
  • Tested applications

    Suitable for: Flow Cyt (Intra), WB, IHC-P, ICC/IFmore details
  • Species reactivity

    Reacts with: Human
    Does not react with: Mouse, Rat
  • Immunogen

    Synthetic peptide within Human SHP1 aa 1-100. The exact sequence is proprietary.
    Database link: P29350

  • Positive control

    • IHC-P: Human tonsil tissue. WB: THP1, Jurkat, Raji and lymph node lysates. ICC: Raji cells. Flow Cyt (intra): Jurkat cells.
  • General notes

    This product is a recombinant monoclonal antibody, which offers several advantages including:

    • - High batch-to-batch consistency and reproducibility
    • - Improved sensitivity and specificity
    • - Long-term security of supply
    • - Animal-free production
    For more information see here.

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

Properties

  • Form

    Liquid
  • Storage instructions

    Shipped at 4°C. Store at -20°C. Stable for 12 months at -20°C.
  • Dissociation constant (KD)

    KD = 4.95 x 10 -11 M
    Learn more about KD
  • Storage buffer

    pH: 7.2
    Preservative: 0.01% Sodium azide
    Constituents: 9% PBS, 40% Glycerol (glycerin, glycerine), 0.05% BSA, 50% Tissue culture supernatant
  • Concentration information loading...
  • Purity

    Tissue culture supernatant
  • Clonality

    Monoclonal
  • Clone number

    EPR5519
  • Isotype

    IgG
  • Research areas

    • Cell Biology
    • Cell Cycle
    • Kinases/Phosphatases
    • Phosphatases
    • Signal Transduction
    • Signaling Pathway
    • G Protein Signaling
    • GPCR
    • Cancer
    • Cell cycle
    • Kinases/phosphatases
    • Phosphatases
    • Cancer
    • Signal transduction
    • G protein signaling
    • GPCR
    • Developmental Biology
    • Organogenesis
    • Hematopoietic system development

Associated products

  • Alternative Versions

    • PE Anti-SHP1 antibody [EPR5519] (ab209002)
    • Alexa Fluor® 488 Anti-SHP1 antibody [EPR5519] (ab211381)
    • Anti-SHP1 antibody [EPR5519] - BSA and Azide free (ab240949)
  • Compatible Secondaries

    • Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077)
    • Goat Anti-Rabbit IgG H&L (HRP) (ab205718)
  • Isotype control

    • Rabbit IgG, monoclonal [EPR25A] - Isotype Control (ab172730)
  • Positive Controls

    • Jurkat cytoplasmic extract lysate (ab14842)
  • Recombinant Protein

    • Recombinant human SHP1 protein (ab51289)

Applications

The Abpromise guarantee

Our Abpromise guarantee covers the use of ab124942 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
Flow Cyt (Intra)
1/10 - 1/100.

ab172730 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody.  

WB
1/1000 - 1/10000. Predicted molecular weight: 68 kDa.
IHC-P
1/1000 - 1/2000. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.
ICC/IF
1/250 - 1/500.
Notes
Flow Cyt (Intra)
1/10 - 1/100.

ab172730 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody.  

WB
1/1000 - 1/10000. Predicted molecular weight: 68 kDa.
IHC-P
1/1000 - 1/2000. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.
ICC/IF
1/250 - 1/500.

Target

  • Function

    Plays a key role in hematopoiesis. This PTPase activity may directly link growth factor receptors and other signaling proteins through protein-tyrosine phosphorylation. The SH2 regions may interact with other cellular components to modulate its own phosphatase activity against interacting substrates. Together with MTUS1, induces UBE2V2 expression upon angiotensin II stimulation.
  • Tissue specificity

    Isoform 1 is expressed in hematopoietic cells. Isoform 2 is expressed in non-hematopoietic cells.
  • Sequence similarities

    Belongs to the protein-tyrosine phosphatase family. Non-receptor class 2 subfamily.
    Contains 2 SH2 domains.
    Contains 1 tyrosine-protein phosphatase domain.
  • Post-translational
    modifications

    Phosphorylated on serine and tyrosine residues.
  • Cellular localization

    Cytoplasm. Nucleus. In neurons, translocates into the nucleus after treatment with angiotensin II.
  • Target information above from: UniProt accession P29350 The UniProt Consortium
    The Universal Protein Resource (UniProt) in 2010
    Nucleic Acids Res. 38:D142-D148 (2010) .

    Information by UniProt
  • Database links

    • Entrez Gene: 5777 Human
    • Omim: 176883 Human
    • SwissProt: P29350 Human
    • Unigene: 63489 Human
    • Alternative names

      • 70Z-SHP antibody
      • EC 3.1.3.48 antibody
      • HCP antibody
      • HCPH antibody
      • Hematopoietic cell phosphatase antibody
      • Hematopoietic cell protein tyrosine phosphatase antibody
      • Hematopoietic cell protein-tyrosine phosphatase antibody
      • HPTP1C antibody
      • Protein tyrosine phosphatase 1C antibody
      • Protein tyrosine phosphatase non receptor type 6 antibody
      • Protein tyrosine phosphatase SHP1 antibody
      • Protein-tyrosine phosphatase 1C antibody
      • protein-tyrosine phosphatase SHP 1 antibody
      • Protein-tyrosine phosphatase SHP-1 antibody
      • PTN6_HUMAN antibody
      • PTP 1C antibody
      • PTP-1C antibody
      • PTP1C antibody
      • Ptpn6 antibody
      • SH PTP 1 antibody
      • SH PTP1 antibody
      • SH-PTP1 antibody
      • SHP 1 antibody
      • SHP 1L antibody
      • SHP1 antibody
      • SHP1L antibody
      • tyrosine protein phosphatase non receptor type 6 antibody
      • Tyrosine-protein phosphatase non-receptor type 6 antibody
      see all

    Images

    • Western blot - Anti-SHP1 antibody [EPR5519] (ab124942)
      Western blot - Anti-SHP1 antibody [EPR5519] (ab124942)
      All lanes : Anti-SHP1 antibody [EPR5519] (ab124942) at 1/1000 dilution

      Lane 1 : Wild-type THP-1 cell lysate
      Lane 2 : PTPN6 knockout THP-1 cell lysate
      Lane 3 : Jurkat cell lysate
      Lane 4 : K562 cell lysate

      Lysates/proteins at 20 µg per lane.

      Performed under reducing conditions.

      Predicted band size: 68 kDa
      Observed band size: 70 kDa why is the actual band size different from the predicted?



      Lanes 1 - 4: Merged signal (red and green). Green - ab124942 observed at 70 kDa. Red - loading control ab8245 (Mouse anti-GAPDH antibody [6C5]) observed at 37 kDa.

      ab124942 was shown to react with SHP1 in wild-type THP-1 cells in Western blot with loss of signal observed in PTPN6 knockout sample. Wild-type THP-1 and PTPN6 knockout cell lysates were subjected to SDS-PAGE. Membranes were blocked in 3 % milk in TBS-T (0.1 % Tween®) before incubation with ab124942 and ab8245 (Mouse anti-GAPDH antibody [6C5]) overnight at 4 °C at a 1 in 1000 dilution and a 1 in 20000 dilution respectively. Blots were incubated with Goat anti-Rabbit IgG H&L (IRDye® 800CW) preabsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye® 680RD) preabsorbed (ab216776) secondary antibodies at 1 in 20000 dilution for 1 h at room temperature before imaging.

    • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-SHP1 antibody [EPR5519] (ab124942)
      Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-SHP1 antibody [EPR5519] (ab124942)

      ab124942, at a 1/1000 dilution, staining SHP1 in paraffin-embedded Human tonsil tissue by immunohistochemistry

      Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.

    • Immunocytochemistry/ Immunofluorescence - Anti-SHP1 antibody [EPR5519] (ab124942)
      Immunocytochemistry/ Immunofluorescence - Anti-SHP1 antibody [EPR5519] (ab124942)

      Immunocytochemistry/Immunofluorescence analysis of Raji (human Burkitt's lymphoma) cells labelling SHP1 with purified ab124942 at a dilution of 1/500. Cells were fixed with 4% Paraformaldehyde and permeabilised with 0.1% tritonX-100. An Alexa Fluor® 488-conjugated goat anti-rabbit IgG (ab150077) at dilution of 1/1000 was used as the secondary antibody. Nuclei counterstained with DAPI (blue).

      Secondary Only Control: PBS was used instead of the primary antibody as the negative control.

    • Western blot - Anti-SHP1 antibody [EPR5519] (ab124942)
      Western blot - Anti-SHP1 antibody [EPR5519] (ab124942)
      All lanes : Anti-SHP1 antibody [EPR5519] (ab124942) at 1/1000 dilution

      Lane 1 : Jurkat cell lysate
      Lane 2 : Raji cell lysate
      Lane 3 : lymph node lysate

      Lysates/proteins at 10 µg per lane.

      Secondary
      All lanes : standard HRP labelled goat anti-rabbit at 1/2000 dilution

      Predicted band size: 68 kDa

    • Flow Cytometry (Intracellular) - Anti-SHP1 antibody [EPR5519] (ab124942)
      Flow Cytometry (Intracellular) - Anti-SHP1 antibody [EPR5519] (ab124942)

      Intracellular Flow Cytometry analysis of Jurkat (Human T cell leukemia T lymphocyte) cells labeling SHP1 with purified ab124942 at 1/100 dilution (10µg/mL) (Red). Cells were fixed with 4% Paraformaldehyde and permeabilised with 90% Methanol. A Goat anti rabbit IgG (Alexa Fluor® 488, ab150077) secondary antibody was used at 1/2000. Isotype control - Rabbit monoclonal IgG (Black). Unlabeled control - Cells without incubation with primary antibody and secondary antibody (Blue). 

       

       

    • OI-RD Scanning - Anti-SHP1 antibody [EPR5519] (ab124942)
      OI-RD Scanning - Anti-SHP1 antibody [EPR5519] (ab124942)
      Equilibrium disassociation constant (KD)
      Learn more about KD

      Click here to learn more about KD
    • Anti-SHP1 antibody [EPR5519] (ab124942)
      Anti-SHP1 antibody [EPR5519] (ab124942)

    Protocols

    • Flow cytometry protocols
    • Immunohistochemistry protocols
    • Immunocytochemistry & immunofluorescence protocols
    • Western blot protocols

    Click here to view the general protocols

    Datasheets and documents

    • SDS download

    • Datasheet download

      Download

    References (7)

    Publishing research using ab124942? Please let us know so that we can cite the reference in this datasheet.

    ab124942 has been referenced in 7 publications.

    • Bian Y  et al. SMURF1-mediated ubiquitylation of SHP-1 promotes cell proliferation and invasion of endometrial stromal cells in endometriosis. Ann Transl Med 9:362 (2021). PubMed: 33842583
    • Chen X  et al. Ropivacaine inhibits cervical cancer cell growth via suppression of the miR-96/MEG2/pSTAT3 axis. Oncol Rep 43:1659-1668 (2020). PubMed: 32323811
    • Cui X  et al. FCRL3 promotes IL-10 expression in B cells through the SHP-1 and p38 MAPK signaling pathways. Cell Biol Int N/A:N/A (2020). PubMed: 32374464
    • Koh JS  et al. Inhibition of STAT3 in gastric cancer: role of pantoprazole as SHP-1 inducer. Cell Biosci 8:50 (2018). PubMed: 30202514
    • Yao D  et al. CD47 is associated with the up-regulation of the PD-1 oncogenic signaling pathway. Int J Clin Exp Pathol 11:5612-5621 (2018). PubMed: 31949648
    • Gladkikh AA  et al. Comparison of the mRNA expression profile of B-cell receptor components in normal CD5-high B-lymphocytes and chronic lymphocytic leukemia: a key role of ZAP70. Cancer Med 6:2984-2997 (2017). PubMed: 29125235
    • Hagberg N  et al. Systemic lupus erythematosus immune complexes increase the expression of SLAM family members CD319 (CRACC) and CD229 (LY-9) on plasmacytoid dendritic cells and CD319 on CD56(dim) NK cells. J Immunol 191:2989-98 (2013). Human . PubMed: 23956418

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