Anti-PIRB antibody [EPR24885-18]

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-PIRB antibody [EPR24885-18] (AB284407)

Immunohistochemical analysis of paraffin-embedded Mouse cerebrum tissue labelling PIRB with ab284407 at 1/500 (6.08 ug/ml) followed by a ready to use Leica DS9800 (BOND™, Polymer Refine Detection) was used. Negative control : No staining on mouse cerebrum. The section was incubated with ab284407 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 Leica DS9800 (BOND™, Polymer Refine Detection) was used.

Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution 2) for 20 mins

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

• IHC-P

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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-PIRB antibody [EPR24885-18] (AB284407)

Immunohistochemical analysis of paraffin-embedded Mouse liver tissue labelling PIRB with ab284407 at 1/500 (6.08 ug/ml) followed by a ready to use Leica DS9800 (BOND^®, Polymer Refine Detection) was used. Positive staining on kuffer cells of mouse liver (PMID : 9054430). The section was incubated with ab284407 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 Leica DS9800 (BOND^®, Polymer Refine Detection) was used.

Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution 2) for 20 mins

• IHC-P

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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-PIRB antibody [EPR24885-18] (AB284407)

Immunohistochemical analysis of paraffin-embedded Mouse spleen tissue labelling PIRB with ab284407 at 1/500 (6.08 ug/ml) followed by a ready to use Leica DS9800 (BOND™ , Polymer Refine Detection) was used. Positive staining on mouse spleen (PMID : 9054430). The section was incubated with ab284407 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 Leica DS9800 (BOND™ , Polymer Refine Detection) was used.

Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution 2) for 20 mins

• IHC-Fr

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Immunohistochemistry (Frozen sections) - Anti-PIRB antibody [EPR24885-18] (AB284407)

Immunohistochemical analysis of 4% PFA-fixed, 0.2% Triton X-100 permeabilized frozen Mouse spleen (fresh) tissue labeling PIRB with ab284407 at 1/100 (6.08 ug/ml) dilution followed by ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor^® 488) preadsorbed at 1/1000 (2 ug/ml) dilution (Green). Positive staining mainly in red pulp of spleen. is observed. The nuclear counterstain was DAPI (Blue).

Secondary antibody control : Secondary antibody is ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor^® 488) preadsorbedat 1/1000 (2 ug/ml) dilution.

• ICC/IF

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Immunocytochemistry/ Immunofluorescence - Anti-PIRB antibody [EPR24885-18] (AB284407)

Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% TritonX-100 permeabilized Mouse spleen cell cells labelling PIRB with ab284407 at 1/50 (12.16 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 cytoplasmic staining in subsets of Mouse spleen cell.

Negative control : EL4 (PMID : 21555536)

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 (2 ug/ml) dilution.

• IP

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Immunoprecipitation - Anti-PIRB antibody [EPR24885-18] (AB284407)

PIRB was immunoprecipitated from 0.35 mg WEHI-3 (mouse leukemia lymphoblast) whole cell lysate 10ug with ab284407 at 1/30 dilution (2ug in 0.35mg lysates). Western blot was performed on the immunoprecipitate using ab284407 at 1/1000 dilution. VeriBlot for IP secondary antibody(HRP)(ab131366) was used at 1/5000 dilution. Lane 1 : WEHI-3 (mouse leukemia lymphoblast) whole cell lysate 10ug

Lane 2 : ab284407 IP in WEHI-3 whole cell lysate

Lane 3 : Rabbit monoclonal IgG (ab172730) instead of ab284407 in WEHI-3 whole cell lysate

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

Exposure time : 48 seconds

All lanes:

Immunoprecipitation - Anti-PIRB antibody [EPR24885-18] (ab284407)

false

• WB

Lab

Western blot - Anti-PIRB antibody [EPR24885-18] (AB284407)

Blocking and diluting buffer and concentration : 5% NFDM/TBSTNegative control : EL4 (PMID : 21555536)

The antibody detects isoform of PirB around 40kDa which consistent with what has been described in the literature (PMID : 31388006)

Exposure time : 81 seconds

All lanes:

Western blot - Anti-PIRB antibody [EPR24885-18] (ab284407) at 1/1000 dilution

Lane 1:

WEHI-3 (mouse leukemia lymphoblast), whole cell lysate at 60 µg

Lane 2:

EL4 (mouse lymphoma T lymphocyte), whole cell lysate at 60 µg

Secondary

All lanes:

Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated (<a href='/en-us/products/secondary-antibodies/goat-rabbit-igg-h-l-hrp-ab97051'>ab97051</a>) at 1/100000 dilution

Predicted band size: 93 kDa

Observed band size: 125 kDa

false

• WB

Lab

Western blot - Anti-PIRB antibody [EPR24885-18] (AB284407)

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

Exposure time : 114 seconds

All lanes:

Western blot - Anti-PIRB antibody [EPR24885-18] (ab284407) at 1/1000 dilution

Lane 1:

Mouse spleen tissue lysate at 20 µg

Lane 2:

Mouse spleen tissue lysate treated with Protein Deglycosylation MIX at 20 µg

Secondary

All lanes:

Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated (<a href='/en-us/products/secondary-antibodies/goat-rabbit-igg-h-l-hrp-ab97051'>ab97051</a>) at 1/100000 dilution

Predicted band size: 93 kDa

Observed band size: 125 kDa

false