Anti-GPA33 antibody [EPR4240] - BSA and Azide free

• IHC-P

Lab

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-GPA33 antibody [EPR4240] - BSA and Azide free (AB247720)

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human colon cancer tissue sections labeling GPA33 with Purified ab108938 at 1 : 400 dilution (0.25 μg/ml). Perform heat mediated antigen retrieval using ab93684 (Tris/EDTA buffer, pH 9.0). ImmunoHistoProbe one step HRP Polymer (ready to use) was used as the secondary antibody. 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 (ab108938).

• ICC/IF

Lab

Immunocytochemistry/ Immunofluorescence - Anti-GPA33 antibody [EPR4240] - BSA and Azide free (AB247720)

This data was developed using ab108938, the same antibody clone in a different buffer formulation.

Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% TritonX-100 permeabilized LoVo (human colorectal adenocarcinoma epithelial cell) cells labelling GPA33 with ab108938 at 1/50 dilution, followed by Goat Anti-Rabbit IgG H&L (Alexa Fluor^® 488) preadsorbed (ab150081) antibody at 1/1000 dilution (Green).

Confocal image showing membrane and weak cytoplasmic staining in LoVo cell line (shown in green). The counterstain was observed in magenta. Nuclear DNA was labelled with DAPI (shown in blue).

Low expression : Caco2.

Image was taken with a confocal microscope (Leica-Microsystems, TCS SP8).

Anti-alpha Tubulin mouse monoclonal antibody (ab7291) was used to counterstain tubulin at 1/1000 dilution (Magenta). The Nuclear counterstain was DAPI (Blue).

Secondary antibody only control : Secondary antibody is Goat Anti-Rabbit IgG H&L (Alexa Fluor^® 488) preadsorbed (ab150081) at 1/1000 dilution.

• IP

Lab

Immunoprecipitation - Anti-GPA33 antibody [EPR4240] - BSA and Azide free (AB247720)

Purified ab108938 at 1 : 20 dilution (0.5ug) immunoprecipitating GPA33 in HT-29 whole cell lysate.

Lane 1 (input) : HT-29 (Human colorectal adenocarcinoma epithelial cell) whole cell lysate (10μg)

Lane 2 (+) : ab108938 + HT-29 whole cell lysate.

Lane 3 (-) : Rabbit monoclonal IgG (ab172730) instead of ab108938 in HT-29 whole cell lysate.

VeriBlot for IP Detection Reagent (HRP) (ab131366) (1/1000) was used for Western blotting.

Blocking Buffer and concentration : 5% NFDM/TBST.

Diluting buffer and concentration : 5% NFDM/TBST.

Observed band size : 50 kDa

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab108938).

All lanes:

Immunoprecipitation - Anti-GPA33 antibody [EPR4240] (<a href='/en-us/products/primary-antibodies/gpa33-antibody-epr4240-ab108938'>ab108938</a>)

Predicted band size: 36 kDa

false

• WB

Unknown

Western blot - Anti-GPA33 antibody [EPR4240] - BSA and Azide free (AB247720)

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab108938).

All lanes:

Western blot - Anti-GPA33 antibody [EPR4240] (<a href='/en-us/products/primary-antibodies/gpa33-antibody-epr4240-ab108938'>ab108938</a>) at 1/1000 dilution

Lane 1:

Human colon lysates at 20 µg

Lane 2:

HT-29 (Human colorectal adenocarcinoma epithelial cell) whole cell lysates at 20 µg

Lane 3:

MOLT-4 (Human lymphoblastic leukemia T lymphoblast) whole cell lysates at 20 µg

Secondary

All lanes:

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

Predicted band size: 36 kDa

Observed band size: 50 kDa

false

• WB

Lab

Western blot - Anti-GPA33 antibody [EPR4240] - BSA and Azide free (AB247720)

This data was developed using ab108938, the same antibody clone in a different buffer formulation.

Blocking and diluting buffer and concentration : 5% NFDM/TBST.

In Western blot, Anti-GAPDH antibody [EPR16891] - Loading Control (ab181602) staining at 1/200000 dilution.

Low expression : Caco-2.

The identity of the lower MW bands at approximately 25 kDa (in lane 1 and 3) are unknown.

All lanes:

Western blot - Anti-GPA33 antibody [EPR4240] (<a href='/en-us/products/primary-antibodies/gpa33-antibody-epr4240-ab108938'>ab108938</a>) at 1/1000 dilution

Lane 1:

Caco-2 (human colorectal adenocarcinoma epithelial cell) whole cell lysate at 20 µg

Lane 2:

HCT 116 (human colorectal carcinoma epithelial cell) whole cell lysate at 20 µg

Lane 3:

LoVo (human colorectal adenocarcinoma epithelial cell) whole cell lysate at 20 µg

Lane 4:

THP-1 (human monocytic leukemia monocyte) whole cell lysate at 20 µg

Secondary

All lanes:

Western blot - Goat Anti-Rabbit IgG H&L (HRP) (<a href='/en-us/products/secondary-antibodies/goat-rabbit-igg-h-l-hrp-ab97051'>ab97051</a>) at 1/20000 dilution

Observed band size: 35 kDa

false

Exposure time: 92s