Anti-Transglutaminase 2 antibody [EPR28142-86] - BSA and Azide free

• ICC/IF

Lab

Immunocytochemistry/ Immunofluorescence - Anti-Transglutaminase 2 antibody [EPR28142-86] - BSA and Azide free (AB310334)

This data was developed using ab310333, the same antibody clone in a different buffer formulation.

ab310333 was shown to react with TGM2 in wild-type A549 cells in immunocytochemistry with loss of signal observed in TGM2 knockout cell line ab261876. 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 ab310333 at 1/500 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).

These data were 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-Transglutaminase 2 antibody [EPR28142-86] - BSA and Azide free (AB310334)

This data was developed using ab310333, the same antibody clone in a different buffer formulation. Immunohistochemical analysis of paraffin-embedded human placenta tissue labelling Transglutaminase 2 with ab310333 at 1/200 (2.485 ug/ml) followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection). Positive staining on human placenta. The section was incubated with ab310333 for 30 mins at room temperature. The immunostaining was performed on a Leica Biosystems BOND® RX instrument incubate slides with 3% Hydrogen Peroxide for 10 mins at room temperature after secondary antibody incubation to reduce the background. Counterstained with Hematoxylin. Secondary antibody only control : Secondary antibody is a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection). Heat mediated antigen retrieval was performed with Tris-EDTA buffer (pH 9.0, Epitope Retrieval Solution2) for 20 mins.

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Transglutaminase 2 antibody [EPR28142-86] - BSA and Azide free (AB310334)

This data was developed using ab310333, the same antibody clone in a different buffer formulation. Immunohistochemical analysis of paraffin-embedded human colon tissue labelling Transglutaminase 2 with ab310333 at 1/200 (2.485 ug/ml) followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection). Positive staining on human colon (PMID : 34103685). The section was incubated with ab310333 for 30 mins at room temperature. The immunostaining was performed on a Leica Biosystems BOND® RX instrumentIncubate slides with 3% Hydrogen Peroxide for 10 mins at room temperature after secondary antibody incubation to reduce the backgroundIncubate slides with 3% Hydrogen Peroxide for 10 mins at room temperature after secondary antibody incubation to reduce the background. Counterstained with Hematoxylin. Secondary antibody only control : Secondary antibody is a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection). Heat mediated antigen retrieval was performed with Tris-EDTA buffer (pH 9.0, Epitope Retrieval Solution2) for 20 mins.

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Transglutaminase 2 antibody [EPR28142-86] - BSA and Azide free (AB310334)

This data was developed using ab310333, the same antibody clone in a different buffer formulation. Immunohistochemical analysis of paraffin-embedded human gastric carcinoma tissue labelling Transglutaminase 2 with ab310333 at 1/200 (2.485 ug/ml) followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection). Positive staining on human gastric carcinoma (PMID : 32467608). The section was incubated with ab310333 for 30 mins at room temperature. The immunostaining was performed on a Leica Biosystems BOND® RX instrument incubate slides with 3% Hydrogen Peroxide for 10 mins at room temperature after secondary antibody incubation to reduce the background. Counterstained with Hematoxylin. Secondary antibody only control : Secondary antibody is a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection). Heat mediated antigen retrieval was performed with Tris-EDTA buffer (pH 9.0, Epitope Retrieval Solution2) for 20 mins.

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Transglutaminase 2 antibody [EPR28142-86] - BSA and Azide free (AB310334)

This data was developed using ab310333, the same antibody clone in a different buffer formulation. Immunohistochemical analysis of paraffin-embedded Human lung tissue labeling Transglutaminase 2 with ab310333 at 1/200 (2.485 ug/ml) followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection). Positive staining on human lung. The section was incubated with ab310333 for 30 mins at room temperature. The immunostaining was performed on a Leica Biosystems BOND® RX instrument incubate slides with 3% Hydrogen Peroxide for 10 mins at room temperature after secondary antibody incubation to reduce the background. Counterstained with Hematoxylin. Secondary antibody only control : Secondary antibody is a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection). Heat mediated antigen retrieval was performed with Tris-EDTA buffer (pH 9.0, Epitope Retrieval Solution2) for 20 mins.

• ICC/IF

Supplier Data

Immunocytochemistry/ Immunofluorescence - Anti-Transglutaminase 2 antibody [EPR28142-86] - BSA and Azide free (AB310334)

This data was developed using ab310333, the same antibody clone in a different buffer formulation. Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% TritonX-100 permeabilized TGM2 KO A549 (human lung carcinoma epithelial cell)(ab267109) cells labelling Transglutaminase 2 with ab310333 at 1/500 (0.994 ug/ml) dilution, followed by ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed antibody at 1/1000 2ug/ml dilution (Green). Confocal image showing cytoplasmic staining in parental A549 cell line, and no staining in TGM2 KO A549 cell line. Image was taken with a confocal microscope(Leica-Microsystems, TCS SP8). ab195889 Anti-alpha Tubulin mouse monoclonal antibody - Microtubule Marker (Alexa Fluor® 594) was used to counterstain tubulin at 1/200 2.5ug/ml dilution (Red). The Nuclear counterstain was DAPI (Blue). Secondary antibody only control : Secondary antibody is ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed at 1/1000 2ug/ml dilution.

• WB

Supplier Data

Western blot - Anti-Transglutaminase 2 antibody [EPR28142-86] - BSA and Azide free (AB310334)

This data was developed using ab310333, the same antibody clone in a different buffer formulation. Blocking and diluting buffer and concentration : 5% NFDM/TBST Performed under reducing conditions. In Western blot, ab310333 was shown to bind specifically to Transglutaminase 2. A band was observed at 77 kDa in wild-type A549 cell lysates whereas no signal observed at this size in TGM2 knockout cell line ab267109 (knockout cell lysate ab257086). In Western blot, anti-GAPDH antibody (ab181602) loading control staining at 1/200000 dilution. Exposure time : 26 seconds

All lanes:

Western blot - Anti-Transglutaminase 2 antibody [EPR28142-86] (<a href='/en-us/products/primary-antibodies/transglutaminase-2-antibody-epr28142-86-ab310333'>ab310333</a>) at 1/1000 dilution

Lane 1:

Wild-type A549 (human lung carcinoma epithelial cell) whole cell lysate at 20 µg

Lane 2:

TGM2 knockout cell line ab267109 (knockout cell lysate <a href='/en-us/products/unavailable/human-tgm2-transglutaminase-2-knockout-a549-cell-lysate-ab257086'>ab257086</a>) at 20 µg

Lane 3:

HeLa (human cervical adenocarcinoma epithelial cell) whole cell lysate at 20 µg

Lane 4:

Human lung tisue lysate at 20 µg

Lane 5:

Human testis tisue lysate at 20 µg

Secondary

All lanes:

Goat Anti-Rabbit IgG (HRP) with minimal cross-reactivity with human IgG at 1/2000 dilution

Observed band size: 77 kDa

false

Exposure time: 26s

• WB

Lab

Western blot - Anti-Transglutaminase 2 antibody [EPR28142-86] - BSA and Azide free (AB310334)

This data was developed using ab310333, the same antibody clone in a different buffer formulation.

ab310333 was shown to react with TGM2 in wild-type A549 cells in Western blot with loss of signal observed in TGM2 knockout cell line ab261876. Wild-type A549 and TGM2 knockout cell lysates were subjected to SDS-PAGE. Membranes were blocked in 5% milk in TBST for 1 hr before incubation with ab310333 overnight at 4 °C at a 1/1000 dilution. Blots were incubated with secondary antibodies at 0.2 µg/mL before imaging.

These data were 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-Transglutaminase 2 antibody [EPR28142-86] (<a href='/en-us/products/primary-antibodies/transglutaminase-2-antibody-epr28142-86-ab310333'>ab310333</a>) at 1/1000 dilution

Lane 1:

Wild-type A549 lysate at 40 µg

Lane 2:

TGM2 knock-out A549 lysate at 40 µg

false