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.

• IHC-P

Unknown

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).

• ICC/IF

Unknown

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)

Unknown

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

Unknown

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).

• ICC/IF

Supplier Data

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

This data was developed using ab32115, the same antibody clone in a different buffer formulation.

ab32115 was shown to react with PXN in wild-type A-431 cells in immunocytochemistry with loss of signal observed in PXN knockout cell line ab261892. Wild-type and knockout cells were mixed and pelleted at a 1 : 1 ratio on coverslips. The cells were fixed with 4% paraformaldehyde (15 min) then permeabilized with 0.1% Triton X-100 (10min) and then blocked with 1x PBS, 0.01% Triton X-100, 5% BSA, 5% NGS. The cells were then incubated with ab32115 at 1/100 dilution overnight at 4°C followed by a further incubation at room temperature for 1h with a goat anti-rabbit secondary antibody to (Alexa Fluor^® 555) at 0.5 μg/ml. Acquisition of the green (wild-type), red (antibody staining) and far-red (knockout) channels was performed. Representative grayscale images of the red channel are shown. Wild-type and knockout cells are outlined with yellow and magenta dashed line, respectively. Schematic representation of the mosaic strategy used is shown on the bottom-right panel. Image was acquired with a Zeiss(LSM-880).

This data was provided by YCharOS Inc., an open science company with the mission of characterizing commercially available antibody reagents for all human proteins. Abcam and YCharOS are working together to help address the reproducibility crisis by enabling the life science community to better evaluate commercially available antibodies.

• IP

Supplier Data

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

This data was developed using ab32115, the same antibody clone in a different buffer formulation.

Immunoprecipitation of PXN in A-431 cells. Lysates were prepared and immunoprecipitation was performed using 16.7 μg of ab32115 pre-coupled to Protein A beads. Samples were then washed and processed for western blot.

This data was provided by YCharOS Inc., an open science company with the mission of characterizing commercially available antibody reagents for all human proteins. Abcam and YCharOS are working together to help address the reproducibility crisis by enabling the life science community to better evaluate commercially available antibodies.

All lanes:

Immunoprecipitation - Anti-Paxillin antibody [E228] (<a href='/ko/products/primary-antibodies/paxillin-antibody-e228-ab32115'>ab32115</a>) at 16.7 µg

All lanes:

A-431 cell Lysates

Observed band size: 64 kDa

false

• 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='/ko/products/primary-antibodies/paxillin-antibody-e228-ab32115'>ab32115</a>)

Predicted band size: 65 kDa

false

• 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).

• WB

Supplier Data

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

This data was developed using ab32115, the same antibody clone in a different buffer formulation.

ab32115 was shown to react with PXN in wild-type A-431 cells in Western blot with loss of signal observed in PXN knockout cell line ab261892. Wild-type A-431 and PXN knockout cell lysates were subjected to SDS-PAGE. Membranes were blocked in 5% milk in TBST for 1 hr before incubation with ab32115 overnight at 4 °C at a 1/10000 dilution. Blots were incubated with secondary antibodies at 0.2 µg/mL before imaging.

This data was provided by YCharOS Inc., an open science company with the mission of characterizing commercially available antibody reagents for all human proteins. Abcam and YCharOS are working together to help address the reproducibility crisis by enabling the life science community to better evaluate commercially available antibodies.

All lanes:

Western blot - Anti-Paxillin antibody [E228] (<a href='/ko/products/primary-antibodies/paxillin-antibody-e228-ab32115'>ab32115</a>) at 1/10000 dilution

Lane 1:

Wild-type A-431 lysate at 30 µg

Lane 2:

PXN knock-out A-431 lysate at 30 µg

Lane 2:

Western blot - Human PXN (Paxillin) knockout A-431 cell line (<a href='/ko/products/cell-lines/human-pxn-paxillin-knockout-a-431-cell-line-ab261892'>ab261892</a>)

Observed band size: 63 kDa

false

• Dot

Unknown

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).