Anti-CD8 alpha antibody [EPR22483-288]

• mIHC

Lab

Multiplex immunohistochemistry - Anti-CD8 alpha antibody [EPR22483-288] (AB245118)

Multiplex immunohistochemistry analysis of formalin/PFA-fixed paraffin-embedded Human tonsil staining of PVRIG/CD112R with ab322348 at a 1/100 (5.16 µg/ml) dilution, CD19 with ab320735 at 1/2000 (0.255 µg/ml) dilution and CD8A with ab245118 at 1/1000 ( 0.375 µg/ml) dilution.

Panel A : merged staining of anti-PVRIG/CD112R (green; Opal™520), anti-CD8A (magenta; Opal™570) and anti-CD19 (yellow; Opal™690) on human tonsil.
Panel B : anti-PVRIG/CD112R staining memory and effector CD8+ T lymphocytes in human tonsil.
Panel C : anti-CD8A staining CD8+ T lymphocytes in human tonsil.
Panel D : anti-CD19 staining B lymphocytes in human tonsil.
Nuclear DNA was labeled with DAPI (shown in blue). ).

The section was incubated in three rounds of staining : in the order of ab322348, ab245118 and ab320735 for 30 mins at room temperature. Each round was followed by a separate fluorescent tyramide signal amplification system.

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

• mIHC

Lab

Multiplex immunohistochemistry - Anti-CD8 alpha antibody [EPR22483-288] (AB245118)

Multiplex immunohistochemistry analysis of formalin/PFA-fixed paraffin-embedded human liver tissue staining TLR2 with ab325499 at a 1 : 100 (5.0 µg/ml) dilution, ab183322 anti-CD14 used at 1 : 100 (1.22 µg/ml) dilution and ab245118 anti-CD8 alpha used at a 1 : 500 (0.75 µg/ml) dilution.

Panel A : anti-TLR2 (green; Opal™520), anti-CD14 (magenta; Opal™570) and anti-CD8 alpha (yellow; Opal™690) on human liver.
Panel B : anti-TLPR2 staining immune cells in human liver.
Panel C : anti-CD14 staining macrophages in human liver.
Panel D : anti-CD8 alpha staining T lymphocytes in human liver.
Nuclear DNA was labeled with DAPI (shown in blue).

The section was incubated in three rounds of staining : in the order of ab325499, ab183322 and ab245118 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.

Nuclear counter stain with DAPI.

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

• mIHC

Supplier Data

Multiplex immunohistochemistry - Anti-CD8 alpha antibody [EPR22483-288] (AB245118)

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 ab245118 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.

• mIHC

Supplier Data

Multiplex immunohistochemistry - Anti-CD8 alpha antibody [EPR22483-288] (AB245118)

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 ab245118 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 Cyt

Unknown

Flow Cytometry - Anti-CD8 alpha antibody [EPR22483-288] (AB245118)

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.

• IHC-P

Unknown

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD8 alpha antibody [EPR22483-288] (AB245118)

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).

• IHC-P

Unknown

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD8 alpha antibody [EPR22483-288] (AB245118)

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).

• mIHC

Lab

Multiplex immunohistochemistry - Anti-CD8 alpha antibody [EPR22483-288] (AB245118)

Multiplex immunohistochemistry analysis of formalin/PFA-fixed paraffin-embedded Human colon staining of PVRIG/CD112R with ab322348 at a 1/100 (5.16 µg/ml) dilution, CD19 with ab320735 at 1/2000 (0.255 µg/ml) dilution and CD8A with ab245118 at 1/1000 ( 0.375 µg/ml) dilution.

Panel A : merged staining of anti-PVRIG/CD112R (green; Opal™520), anti-CD8A (magenta; Opal™570) and anti-CD19 (yellow; Opal™690) on Human colon.
Panel B : anti-PVRIG/CD112R staining memory and effector CD8+ T lymphocytes in Human colon.
Panel C : anti-CD8A staining CD8+ T lymphocytes in Human colon.
Panel D : anti-CD19 staining B lymphocytes in Human colon.
Nuclear DNA was labelled with DAPI (shown in blue). ).

The section was incubated in three rounds of staining : in the order of ab322348, ab245118 and ab320735 for 30 mins at room temperature. Each round was followed by a separate fluorescent tyramide signal amplification system.

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

• mIHC

Lab

Multiplex immunohistochemistry - Anti-CD8 alpha antibody [EPR22483-288] (AB245118)

Multiplex immunohistochemistry analysis of formalin/PFA-fixed paraffin-embedded Human spleen staining of PVRIG/CD112R with ab322348 at a 1/100 (5.16 µg/ml) dilution, CD19 with ab320735 at 1/2000 (0.255 µg/ml) dilution and CD8A with ab245118 at 1/1000 ( 0.375 µg/ml) dilution.

Panel A : merged staining of anti-PVRIG/CD112R (green; Opal™520), anti-CD8A (magenta; Opal™570) and anti-CD19 (yellow; Opal™690) on human spleen.
Panel B : anti-PVRIG/CD112R staining memory and effector CD8+ T lymphocytes in human spleen.
Panel C : anti-CD8A staining CD8+ T lymphocytes in human spleen.
Panel D : anti-CD19 staining B lymphocytes in human spleen.
Nuclear DNA was labelled with DAPI (shown in blue). ).

The section was incubated in three rounds of staining : in the order of ab322348, ab245118 and ab320735 for 30 mins at room temperature. Each round was followed by a separate fluorescent tyramide signal amplification system.

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

• IP

Unknown

Immunoprecipitation - Anti-CD8 alpha antibody [EPR22483-288] (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.

All lanes:

Immunoprecipitation - Anti-CD8 alpha antibody [EPR22483-288] (ab245118)

Predicted band size: 26 kDa

Observed band size: 26 kDa

false