Anti-CD8 alpha 抗体 [EPR22483-288] - BSA and Azide free (ab245830)

Immunohistochemical analysis of paraffin-embedded human tonsil tissue labeling CD8 alpha with ab245118 at 1/1000 dilution, followed by a ready to use Goat Anti-Rabbit IgG H&L (HRP). Positive staining on the T cell zone of human tonsil (PMID: 16504163) is observed. Counterstained with hematoxylin.
Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is a ready to use Goat Anti-Rabbit IgG H&L (HRP).
Perform heat mediated antigen retrieval using ab93684 (Tris/EDTA buffer, pH 9.0).

Immunohistochemical analysis of paraffin-embedded human colon tissue labeling CD8 alpha with ab245118 at 1/1000 dilution, followed by a ready to use Goat Anti-Rabbit IgG H&L (HRP). Positive staining on lymphocytes of human colon (PMID: 26310568) is observed. Counterstained with hematoxylin.
Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is a ready to use Goat Anti-Rabbit IgG H&L (HRP).
Perform heat mediated antigen retrieval using ab93684 (Tris/EDTA buffer, pH 9.0).

This data was developed using ab245118, the same antibody clone in a different buffer formulation.

Multiplex immunohistochemistry analysis of formalin/PFA-fixed paraffin-embedded Human spleen tissue labeling CD8 alpha with ab245118 at 1/500 dilution, CD4 with ab238798 at 1/500, and CD19 with ab237772 at 1/5000 dilution.

Panel A: merged staining of anti-CD8 alpha (magenta; Opal™690), anti-CD4 (green; Opal™520) and anti-CD19 (red; Opal™570) on human spleen.
Panel B: anti-CD8 alpha stained on cytotoxic T cells.
Panel C: anti-CD4 stained on T helper cells.
Panel D: anti-CD19 stained on B cells.

Sections  were treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution2) for 20 mins before antibody incubation. The section was incubated in three rounds of staining: in the order of abab245118 for 30 mins, then ab238798 and ab237772 for 10 mins at room temperature. Each round was followed by a separate fluorescent tyramide signal amplification system.

DAPI was used as a nuclear counterstain.

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.

This data was developed using ab245118, the same antibody clone in a different buffer formulation.

Multiplex immunohistochemistry analysis of formalin/PFA-fixed paraffin-embedded Human tonsil tissue labeling CD8 alpha with ab245118 at 1/500 dilution, CD4 with ab238798 at 1/500, and CD19 with ab237772 at 1/5000 dilution.

Panel A: merged staining of anti-CD8 alpha (magenta; Opal™690), anti-CD4 (green; Opal™520) and anti-CD19 (red; Opal™570) on human tonsil.
Panel B: anti-CD8 alpha stained on cytotoxic T cells.
Panel C: anti-CD4 stained on T helper cells.
Panel D: anti-CD19 stained on B cells.

Sections  were treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution2) for 20 mins before antibody incubation. The section was incubated in three rounds of staining: in the order of abab245118 for 30 mins, then ab238798 and ab237772 for 10 mins at room temperature. Each round was followed by a separate fluorescent tyramide signal amplification system.

DAPI was used as a nuclear counterstain.

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.

Flow cytometric analysis of human peripheral blood mononuclear cells (PBMC) labeling CD8 alpha with ab245118 at 1/400 (right panel) compared with a Rabbit monoclonal IgG (ab172730) (left panel). Goat anti rabbit IgG (Dylight^® 488, ab98462) at 1/2000 dilution was used as the secondary antibody.

Cells were stained with rabbit IgG (Left) or anti-CD8 alpha RabMab (Right). Then stained with Alexa Fluor^® 647-conjugated anti-CD4. The expressions of CD4 and CD8 alpha are mutually exclusive.

Gated on viable lymphocytes.

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab245118).

CD8 alpha was immunoprecipitated from 0.35 mg human thymus lysate with ab245118 at 1/30 dilution. Western blot was performed from the immunoprecipitate using ab245118 at 1/500 dilution. VeriBlot for IP Detection Reagent (HRP) (ab131366), was used for detection at 1/1000 dilution.

Lane 1: Human thymus lysate 10 µg (Input).
Lane 2: ab245118 IP in human thymus lysate.
Lane 3: Rabbit monoclonal IgG (ab172730) instead of ab245118 in human thymus lysate.

Blocking/Dilution buffer: 5% NFDM/TBST.
Exposure time: 30 seconds.

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab245118).