Anti-Paxillin antibody [Y113]

8 product images

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  Western blot - Anti-Paxillin antibody [Y113] (ab32084)

  All lanes: Western blot - Anti-Paxillin antibody [Y113] (ab32084) at 1/1000 dilution

  Lane 1: HeLa (Human cervix adenocarcinoma epithelial cell) whole cell lysate

  Lane 2: RAW 264.7 (Mouse Abelson murine leukemia virus-induced tumor macrophage) whole cell lysate

  Lane 3: Mouse heart lysate

  Secondary

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

  Predicted band size: 65 kDa

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  Immunoprecipitation - Anti-Paxillin antibody [Y113] (ab32084)

  Purified ab32084 at 1/20 dilution (0.7μg) immunoprecipitating Paxillin in HeLa whole cell lysate.
  Lane 1 (input): HeLa (Human cervix adenocarcinoma epithelial cell) whole cell lysate 10μg.
  Lane 2 (+): ab32084 + HeLa whole cell lysate.
  Lane 3 (-): Rabbit monoclonal IgG (Rabbit IgG, monoclonal [EPR25A] - Isotype Control ab172730) instead of ab32084 in HeLa whole cell lysate.
  VeriBlot for IP Detection Reagent (HRP) (VeriBlot for IP Detection Reagent (HRP) ab131366) (1/1000 dilution) was used for Western blotting.
  Blocking Buffer and concentration: 5% NFDM/TBST.
  Diluting buffer and concentration: 5% NFDM/TBST.
  Observed band size: 48 kDa

  All lanes: Immunoprecipitation - Anti-Paxillin antibody [Y113] (ab32084)

  Predicted band size: 65 kDa

  Observed band size: 68 kDa

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  Immunocytochemistry/ Immunofluorescence - Anti-Paxillin antibody [Y113] (ab32084)

  Immunocytochemistry analysis of HeLa (Human cervix adenocarcinoma epithelial cell) cells labeling Paxillin with purified ab32084 at 1/50 dilution (2.88 μg/mL). Cells were fixed in 4% Paraformaldehyde and permeabilized with 0.1% tritonX-100. Cells were counterstained with Alexa Fluor® 594 Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker ab195889 Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor^® 594) 1/200 (2.5 μg/mL). Goat anti rabbit IgG (Alexa Fluor^® 488, Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) ab150077) was used as the secondary antibody at 1/1000 (2 μg/mL) dilution. DAPI (blue) was used as nuclear counterstain. PBS instead of the primary antibody was used as the secondary antibody only control.

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  OI-RD Scanning - Anti-Paxillin antibody [Y113] (ab32084)

  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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  Western blot - Anti-Paxillin antibody [Y113] (ab32084)

  Anti-PXN antibody [Y113] (ab32084) staining at 1/1000 dilution, shown in green; Mouse anti-GAPDH antibody [6C5] (Anti-GAPDH antibody [6C5] - Loading Control ab8245) loading control staining at 1/20000 dilution, shown in red. In Western blot, ab32084 was shown to bind specifically to PXN. A band was observed at 70 kDa in wild-type A431 cell lysates with no signal observed at this size in PXN knockout cell line Human PXN (Paxillin) knockout A-431 cell line ab261892 (knockout cell lysate Human PXN (Paxillin) knockout A-431 cell lysate ab261701). To generate this image, wild-type and PXN knockout A431 cell lysates were analysed. First, samples were run on an SDS-PAGE gel then transferred onto a nitrocellulose membrane. Membranes were blocked in 3 % milk in TBS-0.1 % Tween$®$ 20 (TBS-T) before incubation with primary antibodies overnight at 4 °C. Blots were washed four times in TBS-T, incubated with secondary antibodies for 1 h at room temperature, washed again four times then imaged. Secondary antibodies used were Goat anti-Rabbit IgG H&L 800CW and Goat anti-Mouse IgG H&L 680RD at 1/20000 dilution

  All lanes: Western blot - Anti-Paxillin antibody [Y113] (ab32084) at 1/1000 dilution

  Lane 2: Western blot - Human PXN (Paxillin) knockout A-431 cell lysate (Human PXN (Paxillin) knockout A-431 cell lysate ab261701)

  Lane 2: Western blot - Human PXN (Paxillin) knockout A-431 cell line (Human PXN (Paxillin) knockout A-431 cell line ab261892)

  Predicted band size: 65 kDa

  Observed band size: 70 kDa

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  Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Paxillin antibody [Y113] (ab32084)

  Immunohistochemistry analysis of paraffin-embedded human breast carcinoma tissue sections labelling Paxillin with ab32084 at 1/1200 dilution. The section was incubated with ab32084 for 30 mins at room temperature. Rabbit specific IHC polymer detection kit HRP/DAB (Rabbit specific IHC polymer detection kit HRP/DAB ab209101) was used as the secondary antibody. Sections were counterstained with Hematoxylin. Antigen retrieval was heat mediated antigen retrieval with Citrate buffer (pH 6.0, epitope retrieval solution 1) for 20 mins.
  Positive staining on human breast carcinoma tissue. The immunostaining was performed on a Leica Biosystems BOND^® RX instrument.

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  Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Paxillin antibody [Y113] (ab32084)

  Immunohistochemistry analysis of paraffin-embedded human cerebrum tissue sections labelling Paxillin with ab32084 at 1/1200 dilution. The section was incubated with ab32084 for 30 mins at room temperature. Rabbit specific IHC polymer detection kit HRP/DAB (Rabbit specific IHC polymer detection kit HRP/DAB ab209101) was used as the secondary antibody. Sections were counterstained with Hematoxylin. Antigen retrieval was heat mediated antigen retrieval with Citrate buffer (pH 6.0, epitope retrieval solution 1) for 20 mins.
  Positive staining on human cerebrum tissue. The immunostaining was performed on a Leica Biosystems BOND^® RX instrument.

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  Flow Cytometry (Intracellular) - Anti-Paxillin antibody [Y113] (ab32084)

  Intracellular Flow Cytometry analysis ofHeLa (human cervix adenocarcinoma) cells labeling with purified ab32084 at 1/100 dilution (10μg/ml) (Red). Cells were fixed with4% paraformaldehydeand permeabilised with 90% methanol. A Goat anti rabbit IgG (Alexa Fluor^® 488) (Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) ab150077) (1/2000 dilution) was used as the secondary antibody.Rabbit monoclonal IgG (Black) (Rabbit IgG, monoclonal [EPR25A] - Isotype Control ab172730) was used as a isotype control.Cell without incubation with primary antibody and secondary antibody (Blue) were used as unlabeled control.