Anti-SHP1 antibody [EPR5519]

7 product images

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  Western blot - Anti-SHP1 antibody [EPR5519] (ab124942)

  Lanes 1 - 4: Merged signal (red and green). Green - ab124942 observed at 70 kDa. Red - loading control Anti-GAPDH antibody [6C5] - 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 Anti-GAPDH antibody [6C5] - Loading Control 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 (Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed ab216773) and Goat anti-Mouse IgG H&L (IRDye^® 680RD) preabsorbed (Goat anti-Mouse IgG H&L (IRDye® 680RD) preadsorbed ab216776) secondary antibodies at 1 in 20000 dilution for 1 h at room temperature before imaging.

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

  Lane 1: Wild-type THP-1 cell lysate at 20 µg

  Lane 2: PTPN6 knockout THP-1 cell lysate at 20 µg

  Lane 2: Western blot - Human PTPN6 knockout THP-1 cell line (Human PTPN6 knockout THP-1 cell line ab273717)

  Lane 3: Jurkat cell lysate at 20 µg

  Lane 4: K562 cell lysate at 20 µg

  Performed under reducing conditions.

  Predicted band size: 68 kDa

  Observed band size: 70 kDa

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  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.

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  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 (Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) 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.

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  Western blot - Anti-SHP1 antibody [EPR5519] (ab124942)

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

  Lane 1: Jurkat cell lysate at 10 µg

  Lane 2: Raji cell lysate at 10 µg

  Lane 3: lymph node lysate at 10 µg

  Secondary

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

  Predicted band size: 68 kDa

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  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, Goat Anti-Rabbit IgG H&L (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).
  

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  OI-RD Scanning - Anti-SHP1 antibody [EPR5519] (ab124942)

  We have systematically measured KD (the equilibrium dissociation constant between the antibody and its antigen), of more than 840 recombinant antibodies to assess not only their individual KD values but also to see the average affinity of antibody.
  Based on the comparison with published literature values for mouse monoclonal antibodies, Recombinant antibodies appear to be on average 1-2 order of magnitude higher affinity.

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  Image collected and cropped by CiteAb under a CC-BY license from the publication

  Western blot - Anti-SHP1 antibody [EPR5519] (ab124942)

  SHP1 western blot using anti-SHP1 antibody [EPR5519] ab124942. Publication image and figure legend from Saidova, A., Potashnikova, D. M., et al., 2017, Cancer Med, PubMed 29125235.

  
  

  ab124942 was used in this publication in western blot. This may not be the same as the application(s) guaranteed by Abcam. For a full list of applications guaranteed by Abcam for ab124942 please see the product overview.

  Protein expression level of BCR components in B‐CLL ZAP70‐high and ZAP70‐low representative cases. (A) Immunoblot of signaling tyrosine kinases in ZAP70‐low sample (sample 1) and ZAP70‐high sample (sample 2), histone H3 was taken as a technical control. (B) Sample 1 (see A) analyzed by flow cytometry. Median fluorescence intensities of Zap70 in B‐CLL and T‐cell populations differ ×2.85‐fold; MFI(Zap70)T‐cells/MFI(Zap70)B‐CLL = 2.85. (C) Sample 2 (see A) analyzed by flow cytometry. Median fluorescence intensities of Zap70 in B‐CLL and T‐cell populations do not differ. MFI(Zap70)T‐cells/MFI(Zap70)B‐CLL = 1.