Anti-CD37 antibody [RM1306]

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD37 antibody [RM1306] (AB323574)

Immunohistochemical analysis of paraffin-embedded Human tonsil tissue labeling CD37 with ab323574 at 1/300 (1.68 ug/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Positive staining on human tonsil.
The image on (A) ab323574 is applied with Anti-CD37 antibody at 1/300 dilution and (B) ab251820 is applied with Anti-CD37 antibody at 1/50 dilution.
The section was incubated with ab323574 for 30 mins at room temperature.
The immunostaining was performed on a Leica Biosystems BOND® RX instrument

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 Citrate buffer (pH 6.0, Epitope Retrieval Solution 1) for 20 mins

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD37 antibody [RM1306] (AB323574)

Immunohistochemical analysis of paraffin-embedded Human testis tissue labeling CD37 with ab323574 at 1/300 (1.68 ug/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Negative control : no staining on human testis.
The section was incubated with ab323574 for 30 mins at room temperature.
The immunostaining was performed on a Leica Biosystems BOND® RX instrument

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 Citrate buffer (pH 6.0, Epitope Retrieval Solution 1) for 20 mins

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD37 antibody [RM1306] (AB323574)

Immunohistochemical analysis of paraffin-embedded Human spleen tissue labeling CD37 with ab323574 at 1/300 (1.68 ug/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Positive staining on human spleen.
The image on (A) ab323574 is applied with Anti-CD37 antibody at 1/300 dilution and (B) ab251820 is applied with Anti-CD37 antibody at 1/50 dilution.
The section was incubated with ab323574 for 30 mins at room temperature.
The immunostaining was performed on a Leica Biosystems BOND® RX instrument

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 Citrate buffer (pH 6.0, Epitope Retrieval Solution 1) for 20 mins

• Flow Cyt

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Flow Cytometry - Anti-CD37 antibody [RM1306] (AB323574)

Flow cytometric analysis of MCF7 (human breast adenocarcinoma epithelial cell, Left) / Daudi (human Burkitt's lymphoma lymphoblast, Right) cells labelling CD37 with ab323574 at 1/500 dilution (0.1 ug) (Red) compared with a Rabbit monoclonal IgG (ab172730) / Left isotype control and an unlabelled control (cells without incubation with primary antibody and secondary antibody).

Goat Anti-Rabbit IgG (Alexa Fluor® 488, ab150081) at 1/5000 dilution was used as the secondary antibody.

Low expression : MCF7.

• IP

Supplier Data

Immunoprecipitation - Anti-CD37 antibody [RM1306] (AB323574)

CD37 was immunoprecipitated from 0.35 mg Daudi (human Burkitt's lymphoma lymphoblast) whole cell lysate with ab323574 at 1/1000 dilution (2ug in 0.35mg lysates). Western blot was performed on the immunoprecipitate using ab323574 at 1/1000 dilution.

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

All lanes:

Immunoprecipitation - Anti-CD37 antibody [RM1306] (ab323574) at 1/1000 dilution

Lane 1:

Daudi (human Burkitt's lymphoma lymphoblast) whole cell lysate at 10 µg

Lane 2:

ab323574 at 1/30 IP in Daudi (human Burkitt's lymphoma lymphoblast) whole cell lysate at 10 µg

Lane 3:

Rabbit monoclonal IgG (<a href='/en-us/products/primary-antibodies/rabbit-igg-monoclonal-epr25a-isotype-control-ab172730'>ab172730</a>) instead of ab323574 in Daudi whole cell lysate at 10 µg

Secondary

All lanes:

Immunoprecipitation - VeriBlot for IP Detection Reagent (HRP) (<a href='/en-us/products/reagents/veriblot-for-ip-detection-reagent-hrp-ab131366'>ab131366</a>) at 1/5000 dilution

Observed band size: 40 kDa

false

Exposure time: 6s

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD37 antibody [RM1306] (AB323574)

Immunohistochemical analysis of paraffin-embedded Mouse skeletal muscle tissue labeling CD37 with ab323574 at 1/1200 (0.42 ug/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Negative control : no staining on mouse skeletal muscle.
The section was incubated with ab323574 for 30 mins at room temperature.
The immunostaining was performed on a Leica Biosystems BOND® RX instrument

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 Citrate buffer (pH 6.0, Epitope Retrieval Solution 1) for 20 mins

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD37 antibody [RM1306] (AB323574)

Immunohistochemical analysis of paraffin-embedded Mouse spleen tissue labeling CD37 with ab323574 at 1/1200 (0.42 ug/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Positive staining on mouse spleen.
The section was incubated with ab323574 for 30 mins at room temperature.
The immunostaining was performed on a Leica Biosystems BOND® RX instrument

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 Citrate buffer (pH 6.0, Epitope Retrieval Solution 1) for 20 mins

• ICC/IF

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Immunocytochemistry/ Immunofluorescence - Anti-CD37 antibody [RM1306] (AB323574)

Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% Tween-20 permeabilized A20 (mouse reticulum sarcoma B lymphocyte) cells labelling CD37 with ab323574 at 1/50 (10.08 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 membranous staining in A20 cells (shown in green). The counterstain was observed in magenta. Nuclear DNA was labelled with DAPI (shown in blue).
Low expression : 4T1.
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 (Magenta). 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.

• Flow Cyt

Supplier Data

Flow Cytometry - Anti-CD37 antibody [RM1306] (AB323574)

Flow cytometric analysis of 4T1 (mouse mammary gland carcinoma epithelial cell, Left) / A20 (mouse reticulum sarcoma B lymphocyte, Right) cells labelling CD37 with ab323574 at 1/500 dilution (0.1 ug) (Red) compared with a Rabbit monoclonal IgG (ab172730) / Left isotype control and an unlabelled control (cells without incubation with primary antibody and secondary antibody).

Goat Anti-Rabbit IgG (Alexa Fluor® 488, ab150081) at 1/5000 dilution was used as the secondary antibody.

Negative control : 4T1.

• IP

Supplier Data

Immunoprecipitation - Anti-CD37 antibody [RM1306] (AB323574)

CD37 was immunoprecipitated from 0.35 mg A20 (mouse reticulum sarcoma B lymphocyte) whole cell lysate with ab323574 at 1/1000 dilution (2ug in 0.35mg lysates). Western blot was performed on the immunoprecipitate using ab323574 at 1/1000 dilution.

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

All lanes:

Immunoprecipitation - Anti-CD37 antibody [RM1306] (ab323574) at 1/1000 dilution

Lane 1:

A20 (mouse reticulum sarcoma B lymphocyte) whole cell lysate at 10 µg

Lane 2:

ab323574 at 1/30 IP in A20 (mouse reticulum sarcoma B lymphocyte) whole cell lysate at 10 µg

Lane 3:

Rabbit monoclonal IgG (<a href='/en-us/products/primary-antibodies/rabbit-igg-monoclonal-epr25a-isotype-control-ab172730'>ab172730</a>) instead of ab323574 in A20 whole cell lysate at 10 µg

Secondary

All lanes:

Immunoprecipitation - VeriBlot for IP Detection Reagent (HRP) (<a href='/en-us/products/reagents/veriblot-for-ip-detection-reagent-hrp-ab131366'>ab131366</a>) at 1/5000 dilution

Observed band size: 40 kDa

false

Exposure time: 6s

• WB

Supplier Data

Western blot - Anti-CD37 antibody [RM1306] (AB323574)

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

Negative control : testis.

Samples are non-boiled as boiling may cause protein aggregation.

The identity of the bands between 50 kDa and 100 kDa are unknown.

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

All lanes:

Western blot - Anti-CD37 antibody [RM1306] (ab323574) at 1/1000 dilution

Lane 1:

Human tonsil tissue lysate at 20 µg

Lane 2:

Human testis tissue 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 1/100000 dilution

Observed band size: 40 kDa,36 kDa

false

Exposure time: 37s

• WB

Supplier Data

Western blot - Anti-CD37 antibody [RM1306] (AB323574)

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

Negative control : MCF7 (PMID : 33652767), HepG2, heart.

Samples are non-boiled as boiling may cause protein aggregation.

The identity of the higher MW band at approximately 75 kDa (in lane 6) is unknown.

The molecular weight observed is consistent with what has been described in the literature (PMID : 33652767, PMID : 23267131, PMID : 38846084).

This blot of lane 5-6 was developed using a high-sensitivity ECL substrate, allowing for the detection of proteins in the mid-femtogram range.

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

Exposure time : Lane 1-4 : 81 seconds Lane 5-6 : 92 seconds

All lanes:

Western blot - Anti-CD37 antibody [RM1306] (ab323574) at 1/1000 dilution

Lane 1:

Daudi (human Burkitts lymphoma lymphoblast) whole cell lysate at 20 µg

Lane 2:

Raji (human Burkitts lymphoma B lymphocyte) whole cell lysate at 20 µg

Lane 3:

MCF7 (human breast adenocarcinoma epithelial cell) whole cell lysate at 20 µg

Lane 4:

HepG2 (human hepatocellular carcinoma epithelial cell) whole cell lysate at 20 µg

Lane 5:

Mouse spleen tissue lysate at 20 µg

Lane 6:

Mouse heart tissue 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 1/100000 dilution

Observed band size: 40-50 kDa,36 kDa

false

• WB

Supplier Data

Western blot - Anti-CD37 antibody [RM1306] (AB323574)

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

Low expression : 4T1.

Samples are non-boiled as boiling may cause protein aggregation.

The molecular weight observed is consistent with what has been described in the literature (PMID : 33333768).

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

All lanes:

Western blot - Anti-CD37 antibody [RM1306] (ab323574) at 1/1000 dilution

Lane 1:

A20 (mouse reticulum sarcoma B lymphocyte) whole cell lysate at 20 µg

Lane 2:

WEHI-231 (mouse B cell lymphoma B lymphocyte) whole cell lysate at 20 µg

Lane 3:

4T1 (mouse mammary gland carcinoma epithelial cell) 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 1/100000 dilution

Observed band size: 40-64 kDa,36 kDa

false

Exposure time: 103s