Anti-Paxillin antibody [E228] - BSA and Azide free

• ICC/IF

PubMed

Immunocytochemistry/ Immunofluorescence - Anti-Paxillin antibody [E228] - BSA and Azide free (AB238950)

Immunofluorescence analysis of HeLa cells, staining Paxillin with ab32115.

Cells on the right were treated with MGAT1 shRNA. Cells were fixed with 2% paraformaldehyde, permeabilized using 0.2% Triton-X-100 and blocked by 5% BSA for 1 hour. Cells were incubated with primary antibody (1/400) overnight at 4°C. A FITC-conjugated donkey anti-rabbit IgG (1/500) was used as the secondary antibody.

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab32115).

Image from Beheshti Zavareh R et al. PLoS One. 2012;7(9):e43721. doi: 10.1371/journal.pone.0043721. Epub 2012 Sep 5. Fig 3.; doi:10.1371/journal.pone.0043721; September 5 2012 PLoS ONE 7(9): e43721.

• ICC/IF

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Immunocytochemistry/ Immunofluorescence - Anti-Paxillin antibody [E228] - BSA and Azide free (AB238950)

Immunocytochemistry/ Immunofluorescence analysis of HeLa (Human cervix adenocarcinoma epithelial cell) cells labeling Paxillin with Purified ab32115 at 1 : 100 dilution (1.2 µg/ml). Cells were fixed in 4% Paraformaldehyde and permeabilized with 0.1% tritonX-100. Cells were counterstained with ab195889 Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor^® 594) 1 : 200 (2.5 µg/ml). Goat anti rabbit IgG (Alexa Fluor^® 488, ab150077) was used as the secondary antibody at 1 : 1000 (2 µg/ml) dilution. DAPI nuclear counterstain. PBS instead of the primary antibody was used as the secondary antibody only control.

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab32115).

• Flow Cyt (Intra)

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Flow Cytometry (Intracellular) - Anti-Paxillin antibody [E228] - BSA and Azide free (AB238950)

Intracellular Flow Cytometry analysis of HeLa (Human cervix adenocarcinoma epithelial cell) cells labeling Paxillin with Purified ab32115 at 1/20 dilution (10 μg/ml) (red). Cells were fixed with 4% Paraformaldehyde. A Goat anti rabbit IgG (Alexa Fluor^® 488, ab150077) secondary antibody was used at 1/2000. Isotype control - Rabbit monoclonal IgG (Black). Unlabeled control - Cell without incubation with primary antibody and secondary antibody (Blue).

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab32115).

• ICC/IF

Supplier Data

Immunocytochemistry/ Immunofluorescence - Anti-Paxillin antibody [E228] - BSA and Azide free (AB238950)

Immunocytochemistry/ Immunofluorescence analysis of HeLa (Human epithelial cell line from cervix adenocarcinoma) cells labeling Paxillin with ab32115 at 1/100 (1 μg/ml). Cells were fixed with 4% paraformaldehyde and permeabilized with 0.1% Triton X-100. ab150077, an Alexa Fluor^® 488-conjugated goat anti-rabbit IgG (1/1000, 2 μg/ml) was used as the secondary antibody. The cells were counterstained with ab195889, anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor® 594) at 1/200, 2.5 μg/ml. Nuclei counterstained with DAPI (blue).

Confocal image showing membranous staining on HeLa cells.

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab32115).

• IHC-P

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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Paxillin antibody [E228] - BSA and Azide free (AB238950)

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of Human breast carcinoma tissue sections labeling Paxillin with Purified ab32115 at 1 : 50 dilution (2.34 µg/ml). Heat mediated antigen retrieval was performed using ab93684 (Tris/EDTA buffer, pH 9.0). ImmunoHistoProbe one step HRP Polymer (ready to use)was used. Negative control : PBS instead of the primary antibody. Hematoxylin was used as a counterstain.This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab32115).

• IHC-P

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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Paxillin antibody [E228] - BSA and Azide free (AB238950)

Immunohistochemical analysis of Paxillin expression in paraffin-embedded human colon carcinoma using 1/100 ab32115.

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab32115).

• ELISA

Unknown

ELISA - Anti-Paxillin antibody [E228] - BSA and Azide free (AB238950)

Direct ELISA antigen dose-response curve using ab32115 at 0-1000 ng/mL. Antigen (human Paxillin phospho Y31 peptide/ unmodified peptide) concentration of 1000 ng/mL. An alkaline phosphatase-conjugated goat anti-rabbit IgG (H+L) (1/2500) was used as the secondary antibody.

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab32115).

• IP

Unknown

Immunoprecipitation - Anti-Paxillin antibody [E228] - BSA and Azide free (AB238950)

ab32115 (purified) at 1 : 20 dilution (0.5μg) immunoprecipitating Paxillin in HEK-293 whole cell lysate.
Lane 1 (input) : HEK-293 (Human embryonic kidney epithelial cell) whole cell lysate 10μg
Lane 2 (+) : ab32115 & HEK-293 whole cell lysate
Lane 3 (-) : Rabbit monoclonal IgG (ab172730) instead of ab32115 in HEK-293 whole cell lysate
For western blotting, VeriBlot for IP Detection Reagent (HRP) (ab131366) was used for detection at 1 : 1000 dilution.
Blocking and diluting buffer : 5% NFDM/TBST.

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab32115).

All lanes:

Immunoprecipitation - Anti-Paxillin antibody [E228] (<a href='/en-us/products/primary-antibodies/paxillin-antibody-e228-ab32115'>ab32115</a>)

Predicted band size: 65 kDa

false

• Dot

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Dot Blot - Anti-Paxillin antibody [E228] - BSA and Azide free (AB238950)

Dot blot analysis of Paxillin (pY31) peptide (Lane 1) and Paxillin non-phospho peptide (Lane 2) labelling Paxillin with ab32115 at a dilution of 1/1000. ab97051 (Peroxidase conjugated goat anti-rabbit IgG (H+L)) was used as the secondary antibody at a dilution of 1/100000.

Blocking and dilution buffer : 5% NFDM/TBST.

Exposure time : 3 minutes.

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab32115).