Anti-CD37 抗体 [EPR25397-149] (BSA and Azide free) (ab300412)

All lanes : Anti-CD37 antibody [EPR25397-149] (ab300400) at 1/1000 dilution

Lane 1 : Human tonsil tissue lysate
Lane 2 : Human skeletal muscle tissue lysate
Lane 3 : Raji (human Burkitt's lymphoma B lymphocyte), whole cell lysate
Lane 4 : MCF7 (human breast adenocarcinoma epithelial cell), whole cell lysate

Lysates/proteins at 20 µg per lane.

Secondary
All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/100000 dilution

Predicted band size: 31 kDa
Observed band size: 40 kDa why is the actual band size different from the predicted?

This data was developed using ab300400, the same antibody clone in a different buffer formulation.

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

Exposure time: 

Lane 1-2: 37 seconds
Lane 3-4: 70 seconds

Negative control: skeletal muscle, MCF7 (PMID: 33652767)
Samples are non-boiled as boiling may cause protein aggregation.

This data was developed using ab300400, the same antibody clone in a different buffer formulation. 

Immunohistochemical analysis of human tonsil lebeling CD37 with ab300400 at 1/500 dilution followed by a ready to use LeicaDS9800 (BOND™ Polymer Refine Detection). Positive staining was found in human tonsil (PMID: 25934707).
The section was incubated with ab300400 for 30 mins at room temperature. The immunostaining was performed on a Leica Biosystems BOND^® RX instrument. Counterstained with Hematoxylin. 

Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution2) for 20 mins. 

Secondary antibody only control: PBS was used instead of primary antibody (ab300400) followed by ready to use secondary antibody LeicaDS9800 (BOND™ Polymer Refine Detection). 

Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution2) for 20 mins. 

This data was developed using ab300400, the same antibody clone in a different buffer formulation. 

Immunohistochemical analysis of human spleen lebeling CD37 with ab300400 at 1/500 dilution followed by a ready to use LeicaDS9800 (BOND™ Polymer Refine Detection). Positive staining was found in human spleen (PMID: 25934707). The section was incubated with ab300400 for 30 mins at room temperature. The immunostaining was performed on a Leica Biosystems BOND^® RX instrument. Counterstained with Hematoxylin.

Secondary antibody only control: PBS was used instead of primary antibody (ab300400) followed by ready to use secondary antibody LeicaDS9800 (BOND™ Polymer Refine Detection). 

Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution2) for 20 mins. 

This data was developed using ab300400, the same antibody clone in a different buffer formulation. 

Immunohistochemical analysis of human Hodgkin's lymphoma lebeling CD37 with ab300400 at 1/500 dilution followed by a ready to use LeicaDS9800 (BOND™ Polymer Refine Detection). Positive staining was found in human Hodgkin's lymphoma. The section was incubated with ab300400 for 30 mins at room temperature. The immunostaining was performed on a Leica Biosystems BOND^® RX instrument. Counterstained with Hematoxylin.

Secondary antibody only control: PBS was used instead of primary antibody (ab300400) followed by ready to use secondary antibody LeicaDS9800 (BOND™ Polymer Refine Detection). 

Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution2) for 20 mins. 

This data was developed using ab300400, the same antibody clone in a different buffer formulation. 

Immunohistochemical analysis of human skeletal muscle lebeling CD37 with ab300400 at 1/500 dilution followed by a ready to use LeicaDS9800 (BOND™ Polymer Refine Detection). Negative control: No staining was found on human skeletal muscle.
The section was incubated with ab300400 for 30 mins at room temperature. The immunostaining was performed on a Leica Biosystems BOND^® RX instrument. Counterstained with Hematoxylin. 

Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution2) for 20 mins. 

This data was developed using ab300400, the same antibody clone in a different buffer formulation. 

Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized Raji (human Burkitt's lymphoma B lymphocyte) whole cell lysate labeling CD37 with ab300400 at 1/500 dilution, followed by Goat Anti-Rabbit IgG H&L (Alexa Fluor^® 488) (ab150081) secondary antibody at 1/1000 dilution (green). Confocal image showing cytoplasmic and membranous staining in Raji cell line. Negative control: MCF7 (PMID: 33652767). The nuclear counter stain is DAPI (blue).

Tubulin is detected with ab195889 Anti-alpha Tubulin mouse monoclonal antibody - Microtubule Marker (Alexa Fluor® 594) at 1/200 dilution. 

Control: PBS was used instead of primary antibody followed by secondary antibody.

This data was developed using ab300400, the same antibody clone in a different buffer formulation. 

Flow cytometric analysis of human peripheral blood mononuclear cell (PBMC) labelling CD37 with ab300400 at 1/500 dilution compared with a rabbit monoclonal IgG (ab172730) (Left) isotype control. Goat Anti-Rabbit IgG (Alexa Fluor^® 488, ab150081) was used as the secondary antibody at 1/2000 dilution.  Gated on viable cells.
Cells were stained with rabbit IgG or ab300400. Then stained with anti-CD19 conjugated to Alexa Fluor^® 647.

This data was developed using ab300400, the same antibody clone in a different buffer formulation.

Flow cytometry analysis of human peripheral blood mononuclear cells (PBMC) labelling CD37 with ab300400 at 1/500 dilution (Right) compared with isotype control ab172730 (Left). Gated on viable cells.
Goat Anti-Rabbit IgG (Alexa Fluor® 488, ab150081) was used as a secondary antibody. 

This data was developed using ab300400, the same antibody clone in a different buffer formulation.

Flow cytometry analysis of MCF7 (human breast adenocarcinoma epithelial cell, Left) / Raji (human Burkitt's lymphoma B lymphocyte, Right) labelling CD37 with ab300400 at 1/500 dilution (Right) compared with Rabbit monoclonal IgG (ab172730) / Black isotype control. Cell without incubation with primary antibody and secondary antibody / Blue. Gated on viable cells.
Goat Anti-Rabbit IgG (Alexa Fluor^® 488, ab150081) was used as a secondary antibody. 

This data was developed using ab300400, the same antibody clone in a different buffer formulation. 

CD37 was immunoprecipitated from human tonsil tissue lysate with ab300400 at 1/30 dilution. Western blot was performed from the immunoprecipitate using ab300400 at 1/1000 dilution. VeriBlot for IP secondary antibody (HRP) (ab131366) was used as secondary antibody at 1/5000 dilution. 

Lane 1: Human tonsil tissue lysate 10 μg

Lane 2: ab300400 IP in human tonsil tissue lysate

Lane 3: Rabbit monoclonal IgG (ab172730) instead of ab300400 in Human tonsil tissue lysate

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

Exposure time: 15 seconds.