Anti-CD37 antibody [EPR28769-31]

• Flow Cyt

Supplier Data

Flow Cytometry - Anti-CD37 antibody [EPR28769-31] (AB315346)

Flow cytometric analysis of Mouse splenocytes cells labelling CD37 with ab315346 at 1/50 dilution (1 ug, Right) compared with a Rabbit monoclonal IgG (ab172730) / Left isotype control.

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

Gated on viable cells. Cells are co-stained with CD45R/B220 conjugated to Alexa Fluor®647.

• mIHC

Lab

Multiplex immunohistochemistry - Anti-CD37 antibody [EPR28769-31] (AB315346)

Multiplex immunohistochemistry analysis of formalin/PFA-fixed paraffin-embedded mouse colon tissue staining Glycam1 with ab324912 at a 1/4000 ( 0.126 μg/ml) dilution, ab237721 anti-CD3 used at 1/2000 dilution ( 0.26 μg/ml) and ab315346 anti-CD37 used at 1/2000 dilution (0.257 μg/ml).

Panel A : anti-Glycam1 (green; Opal™520), anti-CD3 (magenta; Opal™690), anti-CD37 (gray; Opal™570) on mouse colon.

Panel B : anti-Glycam1 staining HEV (highly endothelial venules) in mouse colon.

Panel C : anti-CD3 staining T lymphocytes in mouse colon.

Panel D : anti-CD37 staining B lymphocytes in mouse colon.

Nuclear DNA was labeled with DAPI (shown in blue).

The section was incubated in three rounds of staining : in the order of ab324912, ab237721 and ab315346 for 30 mins at room temperature. Each round was followed by a separate fluorescent tyramide signal amplification system.

The immunostaining was performed on a Leica Biosystems BOND® RX instrument with an Opal™ 4-color kit. Image acquisition was performed with Leica SP8 confocal microscope.

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-CD37 antibody [EPR28769-31] (AB315346)

Immunohistochemical analysis of paraffin-embedded Mouse cardiac muscle tissue labeling CD37 with ab315346 at 1/2000 (0.257 ug/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Negative control : No staining on mouse cardiac muscle. The section was incubated with ab315346 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 Tris-EDTA buffer (pH 9.0, Epitope Retrieval Solution2) for 20 mins.

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD37 antibody [EPR28769-31] (AB315346)

Immunohistochemical analysis of paraffin-embedded Mouse Burkitt's lymphoma tissue labeling CD37 with ab315346 at 1/2000 (0.257 ug/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Positive staining on mouse Burkitt's lymphoma. The section was incubated with ab315346 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 Tris-EDTA buffer (pH 9.0, Epitope Retrieval Solution2) for 20 mins.

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD37 antibody [EPR28769-31] (AB315346)

Immunohistochemical analysis of paraffin-embedded Mouse spleen tissue labeling CD37 with ab315346 at 1/2000 (0.257 ug/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Positive staining on mouse spleen (PMID : 19282981). The section was incubated with ab315346 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 Tris-EDTA buffer (pH 9.0, Epitope Retrieval Solution2) for 20 mins.

• ICC/IF

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Immunocytochemistry/ Immunofluorescence - Anti-CD37 antibody [EPR28769-31] (AB315346)

Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% Tween-20 permeabilized mouse splenocytes cells labelling CD37 with ab315346 at 1/500 (1.026 ug/ml) dilution, followed by ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed antibody at 1/1000 (2 ug/ml) dilution (Green).

Confocal image showing membranous staining in subsets of mouse splenocytes, which is co-localized with anti-CD45R. Image was taken with a confocal microscope(Leica-Microsystems, TCS SP8).

Anti-human/mouse CD45R/B220 rat monoclonal antibody (Alexa Fluor® 647) was used to counterstain at 1/100 (5 ug/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 (2 ug/ml) dilution.

• mIHC

Lab

Multiplex immunohistochemistry - Anti-CD37 antibody [EPR28769-31] (AB315346)

Multiplex immunohistochemistry analysis of formalin/PFA-fixed paraffin-embedded mouse lymph node tissue staining Glycam1 with ab324912 at a 1/4000 ( 0.126 μg/ml) dilution, ab237721 anti-CD3 used at 1/2000 dilution ( 0.26 μg/ml) and ab315346 anti-CD37 used at 1/2000 dilution (0.257 μg/ml).

Panel A : anti-Glycam1 (green; Opal™520), anti-CD3 (magenta; Opal™690), anti-CD37 (gray; Opal™570) on mouse lymph node.

Panel B : anti-Glycam1 staining HEV (highly endothelial venules) in mouse lymph node.

Panel C : anti-CD3 staining T lymphocytes in mouse lymph node.

Panel D : anti-CD37 staining B lymphocytes in mouse lymph node.

Nuclear DNA was labeled with DAPI (shown in blue).

The section was incubated in three rounds of staining : in the order of ab324912, ab237721 and ab315346 for 30 mins at room temperature. Each round was followed by a separate fluorescent tyramide signal amplification system.

The immunostaining was performed on a Leica Biosystems BOND® RX instrument with an Opal™ 4-color kit. Image acquisition was performed with Leica SP8 confocal microscope.

Heat mediated antigen retrieval was performed with Tris-EDTA buffer (pH 9.0, Epitope Retrieval Solution2) for 20 mins.

• IP

Supplier Data

Immunoprecipitation - Anti-CD37 antibody [EPR28769-31] (AB315346)

CD37 was immunoprecipitated from 0.35 mg Mouse spleen whole cell lysate with ab315346 at 1/30 dilution (2ug in 0.35mg lysates). Western blot was performed on the immunoprecipitate using ab315346 at 1/1000 dilution. VeriBlot for IP secondary antibody(HRP)(ab131366) was used at 1/5000 dilution.

Lane 1 : Mouse spleen whole cell lysate

Lane 2 : ab315346 IP in Mouse spleen whole cell lysate

Lane 3 : Rabbit monoclonal IgG (ab172730) instead of ab315346 in mouse spleen whole cell lysate

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

All lanes:

Immunoprecipitation - Anti-CD37 antibody [EPR28769-31] (ab315346) at 1/30 dilution

All lanes:

Mouse spleen whole cell lysate

Secondary

All lanes:

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

false

Exposure time: 10s

• WB

Supplier Data

Western blot - Anti-CD37 antibody [EPR28769-31] (AB315346)

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

Negative control : Mouse heart.

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

All lanes:

Western blot - Anti-CD37 antibody [EPR28769-31] (ab315346) at 1/1000 dilution

Lane 1:

Mouse spleen tissue lysate at 20 µg

Lane 2:

Mouse heart tissue lysate at 20 µg

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 1/100000 dilution

Observed band size: 40-60 kDa,36 kDa

false

Exposure time: 37s

• WB

Supplier Data

Western blot - Anti-CD37 antibody [EPR28769-31] (AB315346)

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

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

All lanes:

Western blot - Anti-CD37 antibody [EPR28769-31] (ab315346) at 1/1000 dilution

All lanes:

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

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 1/100000 dilution

Observed band size: 40-60 kDa,36 kDa

false

Exposure time: 37s