Anti-CD8 beta antibody [EPR22331-81] - BSA and Azide free

• Flow Cyt

Unknown

Flow Cytometry - Anti-CD8 beta antibody [EPR22331-81] - BSA and Azide free (AB263956)

Flow Cytometric analysis of mouse thymocytes using ab263946 at a 1/500 dilution. Cells were stained with Rabbit monoclonal IgG (ab172730) (Left) or ab263946 (Right). Then stained with anti-CD4 conjugated to Alexa Fluor^® 647. Secondary used is Goat anti rabbit IgG (Alexa Fluor^® 488, ab150077) at a 1/2000 dilution.

Gated on viable cells.

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab263946).

• IHC-Fr

Supplier Data

Immunohistochemistry (Frozen sections) - Anti-CD8 beta antibody [EPR22331-81] - BSA and Azide free (AB263956)

Immunohistochemical analysis of 4% PFA fixed 0.2% Triton X-100 permeabilized frozen mouse thymus tissue labeling CD8 beta with ab263946 at 1/100 (5.13 µg/ml) dilution followed by ab150077 Alexa Fluor^® 488 Goat anti-Rabbit secondary at 1/1000 (2 µg/ml) dilution. The nuclear counterstain was DAPI (Blue). Positive staining on T cells of mouse thymus (PMID : 12702501) is observed.

Heat mediated antigen retrieval using sodium citrate buffer (10mM citrate pH 6.0 + 0.05% Tween-20).

Secondary antibody only control : Used PBS instead of primary antibody, secondary antibody was ab150077 Alexa Fluor^® 488 Goat anti-Rabbit secondary at 1/1000 (2 µg/ml) dilution.

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab263946).

• IHC-Fr

Supplier Data

Immunohistochemistry (Frozen sections) - Anti-CD8 beta antibody [EPR22331-81] - BSA and Azide free (AB263956)

Immunohistochemical analysis of 4% PFA fixed 0.2% Triton X-100 permeabilized frozen mouse liver tissue labeling CD8 beta with ab263946 at 1/100 (5.13 µg/ml) dilution followed by ab150077 Alexa Fluor^® 488 Goat anti-Rabbit secondary at 1/1000 (2 µg/ml) dilution. The nuclear counterstain was DAPI (Blue). Negative staining on mouse liver.

Negative control : liver (PMID : 3145195).

Heat mediated antigen retrieval using sodium citrate buffer (10mM citrate pH 6.0 + 0.05% Tween-20).

Secondary antibody only control : Used PBS instead of primary antibody, secondary antibody was ab150077 Alexa Fluor^® 488 Goat anti-Rabbit secondary at 1/1000 (2 µg/ml) dilution.

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab263946).

• WB

Lab

Western blot - Anti-CD8 beta antibody [EPR22331-81] - BSA and Azide free (AB263956)

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab263946).Blocking and diluting buffer and concentration : 5% NFDM/TBST.

CD8 beta is a glycoprotein. The molecular weight observed is consistent with what has been described in the literature (PMID : 11719190).

Exposure time :
Lane 1 : 3 secs.
Lane 2 : 3 mins.

All lanes:

Western blot - Anti-CD8 beta antibody [EPR22331-81] (<a href='/en-us/products/primary-antibodies/cd8-beta-antibody-epr22331-81-ab263946'>ab263946</a>) at 1/1000 dilution

Lane 1:

Mouse thymus lysate at 20 µg

Lane 2:

Mouse lymph node 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

Predicted band size: 24 kDa

Observed band size: 28-32 kDa

false

• IP

Supplier Data

Immunoprecipitation - Anti-CD8 beta antibody [EPR22331-81] - BSA and Azide free (AB263956)

CD8 beta was immunoprecipitated from 0.35 mg mouse thymus lysate 10μg with ab263946 at 1/30 dilution (2μg in 0.35mg lysates). Western blot was performed on the immunoprecipitate using ab263946 at 1/1000 dilution. VeriBlot for IP Detection Reagent (HRP) (ab131366) was used at 1/5000 dilution.

Lane 1 : Mouse thymus lysate 10μg
Lane 2 : ab263946 IP in mouse thymus lysate
Lane 3 : Rabbit monoclonal IgG (ab172730) instead of ab263946 in mouse thymus lysate

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

Exposure time : 10 seconds.

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab263946).

All lanes:

Immunoprecipitation - Anti-CD8 beta antibody [EPR22331-81] (<a href='/en-us/products/primary-antibodies/cd8-beta-antibody-epr22331-81-ab263946'>ab263946</a>)

Predicted band size: 24 kDa

false