Anti-Paxillin antibody [Y113] - BSA and Azide free

• Flow Cyt (Intra)

Unknown

Flow Cytometry (Intracellular) - Anti-Paxillin antibody [Y113] - BSA and Azide free (AB216652)

Intracellular Flow Cytometry analysis of HeLa (human cervix adenocarcinoma) cells labeling with purified ab32084 at 1/100 dilution (10ug/ml) (Red). Cells were fixed with 4% paraformaldehyde and permeabilised with 90% methanol. A Goat anti rabbit IgG (Alexa Fluor^® 488) (ab150077) (1/2000 dilution) was used as the secondary antibody. Rabbit monoclonal IgG (Black) (ab172730) was used as a isotype control.Cell without incubation with primary antibody and secondary antibody (Blue) were used as unlabeled control.

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab32084).

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Paxillin antibody [Y113] - BSA and Azide free (AB216652)

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 (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.
This data was developed using ab32084, the same antibody clone in a different buffer formulation.

• ICC/IF

Supplier Data

Immunocytochemistry/ Immunofluorescence - Anti-Paxillin antibody [Y113] - BSA and Azide free (AB216652)

This data was developed using ab32084, the same antibody clone in a different buffer formulation.

ab32084 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 ab32084 at 1/150 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.

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Paxillin antibody [Y113] - BSA and Azide free (AB216652)

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 (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.
This data was developed using ab32084, the same antibody clone in a different buffer formulation.

• ICC/IF

Supplier Data

Immunocytochemistry/ Immunofluorescence - Anti-Paxillin antibody [Y113] - BSA and Azide free (AB216652)

This data was developed using ab32084, the same antibody clone in a different buffer formulation.
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 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 (blue) was used as nuclear counterstain. PBS instead of the primary antibody was used as the secondary antibody only control.

• IP

Lab

Immunoprecipitation - Anti-Paxillin antibody [Y113] - BSA and Azide free (AB216652)

This data was developed using ab32084, the same antibody clone in a different buffer formulation.
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 (ab172730) instead of ab32084 in HeLa whole cell lysate.
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.

All lanes:

Immunoprecipitation - Anti-Paxillin antibody [Y113] (<a href='/ko/products/primary-antibodies/paxillin-antibody-y113-ab32084'>ab32084</a>)

Predicted band size: 65 kDa

Observed band size: 68 kDa

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• IP

Supplier Data

Immunoprecipitation - Anti-Paxillin antibody [Y113] - BSA and Azide free (AB216652)

This data was developed using ab32084, the same antibody clone in a different buffer formulation.

Immunoprecipitation of PXN in A-431 cells. Lysates were prepared and immunoprecipitation was performed using 13.6 μg of ab32084 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 [Y113] (<a href='/ko/products/primary-antibodies/paxillin-antibody-y113-ab32084'>ab32084</a>) at 13.6 µg

All lanes:

A-431 cell Lysates

Observed band size: 64 kDa

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• WB

Supplier Data

Western blot - Anti-Paxillin antibody [Y113] - BSA and Azide free (AB216652)

This data was developed using ab32084, the same antibody clone in a different buffer formulation.

ab32084 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 ab32084 overnight at 4 °C at a 1/1000 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 [Y113] (<a href='/ko/products/primary-antibodies/paxillin-antibody-y113-ab32084'>ab32084</a>) at 1/1000 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

• WB

Lab

Western blot - Anti-Paxillin antibody [Y113] - BSA and Azide free (AB216652)

Anti-PXN antibody [Y113] (ab32084) staining at 1/1000 dilution, shown in green; Mouse anti-GAPDH antibody [6C5] (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 ab261892 (knockout 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] (<a href='/ko/products/primary-antibodies/paxillin-antibody-y113-ab32084'>ab32084</a>) at 1/1000 dilution

Lane 2:

Western blot - Human PXN (Paxillin) knockout A-431 cell lysate (<a href='/ko/products/cell-lysates/human-pxn-paxillin-knockout-a-431-cell-lysate-ab261701'>ab261701</a>)

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>)

Predicted band size: 65 kDa

Observed band size: 70 kDa

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• IHC-P

Unknown

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Paxillin antibody [Y113] - BSA and Azide free (AB216652)

ab32084 (Unpurified format) showing positive staining in Normal ovary tissue.

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab32084).

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

• IHC-P

Unknown

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Paxillin antibody [Y113] - BSA and Azide free (AB216652)

ab32084 (Unpurified format) showing positive staining in Transitional cell carcinoma of kidney tissue.

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab32084).

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

• IHC-P

Unknown

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Paxillin antibody [Y113] - BSA and Azide free (AB216652)

ab32084 (Unpurified format) showing positive staining in Ovarian carcinoma tissue.

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab32084).

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

• WB

Unknown

Western blot - Anti-Paxillin antibody [Y113] - BSA and Azide free (AB216652)

All lanes:

Western blot - Anti-Paxillin antibody [Y113] (<a href='/ko/products/primary-antibodies/paxillin-antibody-y113-ab32084'>ab32084</a>) 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) (<a href='/ko/products/secondary-antibodies/goat-rabbit-igg-h-l-hrp-ab97051'>ab97051</a>) at 1/20000 dilution

Predicted band size: 65 kDa

false

• OI-RD Scanning

Unknown

OI-RD Scanning - Anti-Paxillin antibody [Y113] - BSA and Azide free (AB216652)

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.