Anti-CD8 alpha antibody [EPR21769]

• Flow Cyt

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Flow Cytometry - Anti-CD8 alpha antibody [EPR21769] (AB217344)

Flow cytometric analysis of mouse primary splenocytes labeling CD8 alpha with ab217344 at 1/500 dilution (right panel) compared with a Rabbit IgG, monoclonal [EPR25A] - Isotype Control (ab172730) (left panel). Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077) at 1/2000 dilution was used as the secondary antibody.

Cells were surface stained with CD4-Alexa Fluor^® 647, then stained with rabbit IgG (Left) / ab217344 (Right) separately. CD4 and CD8 alpha are mutually exclusive expressed in mouse spleen. Gated on total viable cells.

• IHC-P

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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD8 alpha antibody [EPR21769] (AB217344)

Immunohistochemical analysis of paraffin-embedded mouse colon tissue labeling CD8 alpha with ab217344 at 1/2000 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) Ready to use. Positive staining on stromal cells of mouse colon. Counter stained with Hematoxylin. Heat mediated antigen retrieval was performed using Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

Secondary antibody only control : Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) Ready to use.

• IHC-Fr

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Immunohistochemistry (Frozen sections) - Anti-CD8 alpha antibody [EPR21769] (AB217344)

Immunohistochemical analysis of 4% paraformaldehyde-fixed, 0.2% Triton X-100 permeabilized frozen mouse thymus tissue labeling CD8 alpha with ab217344 at 1/500 dilution, followed by Goat Anti-Rabbit IgG H&L (Alexa Fluor^® 488) (ab150077) secondary antibody at 1/1000 dilution (green). Positive membrane staining on mouse thymus tissue section (PMID : 25616911).

The nuclear counter stain is DAPI (blue).

Secondary antibody only control : Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (Alexa Fluor^® 488) (ab150077) at 1/1000 dilution.

• IHC-Fr

Supplier Data

Immunohistochemistry (Frozen sections) - Anti-CD8 alpha antibody [EPR21769] (AB217344)

Immunohistochemical analysis of 4% paraformaldehyde-fixed, 0.2% Triton X-100 permeabilized frozen mouse spleen tissue labeling CD8 alpha with ab217344 at 1/500 dilution, followed by Goat Anti-Rabbit IgG H&L (Alexa Fluor^® 488) (ab150077) secondary antibody at 1/1000 dilution (green). Positive membrane staining on mouse spleen (PMID : 25616911).

The nuclear counter stain is DAPI (blue).

Secondary antibody only control : Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (Alexa Fluor^® 488) (ab150077) at 1/1000 dilution.

• IHC-P

Lab

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD8 alpha antibody [EPR21769] (AB217344)

Tissue Microarrays stained for "Anti-CD8 alpha antibody [EPR21769]" using "ab217344"in immunohistochemical analysis. This table provides a detailed overview of positive (tick mark) and negative (cross mark) staining per sample type tested. The sections were pre-treated using Heat mediated antigen retrieval using ab93684 (Tris/EDTA buffer, pH 9.0). The sections were incubated with ab217344 at +4°C overnight followed by a ready to use Goat Anti-Rabbit IgG H&L (HRP polymer).

• IP

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Immunoprecipitation - Anti-CD8 alpha antibody [EPR21769] (AB217344)

CD8 alpha was immunoprecipitated from 0.35 mg of mouse thymus lysate with ab217344 at 1/30 dilution. Western blot was performed from the immunoprecipitate using ab217344 at 1/2000 dilution. VeriBlot for IP Detection Reagent (HRP) (ab131366), was used for detection at 1/10000 dilution.

Lane 1 : Mouse thymus lysate 10 μg (Input).

Lane 2 : ab217344 IP in mouse thymus lysate.

Lane 3 : Rabbit monoclonal IgG (ab172730) instead of ab214344 in mouse thymus lysate.

Exposure time : 5 seconds.

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

The two bands are different isoforms that are consistent with the literature (PMID 3085089).

All lanes:

Immunoprecipitation - Anti-CD8 alpha antibody [EPR21769] (ab217344)

Predicted band size: 26 kDa

Observed band size: 34-38 kDa

false

• WB

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Western blot - Anti-CD8 alpha antibody [EPR21769] (AB217344)

Exposure time : Lane 1 : 3 sconds; Lane 2 : 40 seconds; Lane 3 : 15 seconds.

Blocking/Dilution buffer : 5% NFDM/TBST.

The two bands are different isoforms that are consistent with the literature (PMID 3085089).

The blot was developed on a BIO-RAD® ChemiDoc™ MP instrument.

All lanes:

Western blot - Anti-CD8 alpha antibody [EPR21769] (ab217344) at 1/1000 dilution

Lane 1:

Mouse thymus lysate at 20 µg

Lane 2:

Mouse spleen lysate at 20 µg

Lane 3:

Mouse lymph node lysate at 10 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

Predicted band size: 26 kDa

Observed band size: 35 kDa

true