Anti-PD1 antibody [EPR23119-111] - BSA and Azide free

• IHC-P

Unknown

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-PD1 antibody [EPR23119-111] - BSA and Azide free (AB270753)

Immunohistochemical analysis of paraffin-embedded Human tonsil tissue labeling PD1 with ab243644 at 1/500 followed by a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101). Membranous staining on germinal center of human tonsil (PMID : 29042640, PMID : 16819321) is observed.
The section was incubated with ab243644 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 Rabbit specific IHC polymer detection kit HRP/DAB (ab209101).

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

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab243644).

• IHC-P

Unknown

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-PD1 antibody [EPR23119-111] - BSA and Azide free (AB270753)

Immunohistochemical analysis of paraffin-embedded Human Hodgkin's lymphoma tissue labeling PD1 with ab243644 at 1/500 followed by a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101). Positive staining on human Hodgkin's lymphoma (PMID : 16819321) is observed.
The section was incubated with ab243644 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 Rabbit specific IHC polymer detection kit HRP/DAB (ab209101).

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

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab243644).

• mIHC

Lab

Multiplex immunohistochemistry - Anti-PD1 antibody [EPR23119-111] - BSA and Azide free (AB270753)

This data was developed using ab243644, the same antibody clone in a different buffer formulation. Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of Human tonsil labelling PD1 with ab243644 at 1/500 dilution (1.02 µg/mL) (D), Ki67 with ab16667 at 1/200 dilution (0.15 μg/ml) (B) and PD-L1 with ab228462 at 1/100 dilution (0.52 μg/ml) (C). Opal Polymer HRP Ms + Rb was used as a secondary antibody, and DAPI was used for a nuclear counter stain. Heat mediated antigen retrieval with Citrate buffer (pH 6.0, epitope retrieval solution 1) for 20 mins. Panel A : merged staining of anti-Ki67 (magenta; Opal™690), anti-PD-L1 (red; Opal™570) and anti-PD1 (green; Opal™520) on human tonsil. Panel B : anti-Ki67 stained on nucleus of proliferating cells. Panel C : anti-PD-L1 stained on membrane of cells involved in T cell inhibition. Panel D : anti-PD1 stained on antigen-stimulated T cells. The section was incubated in three rounds of staining : in the order of ab16667 for 10 mins, ab243644 for 30 mins and ab228462 for 10 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.

• IHC-P

Unknown

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-PD1 antibody [EPR23119-111] - BSA and Azide free (AB270753)

Immunohistochemical analysis of paraffin-embedded Human endometrial carcinoma tissue labeling PD1 with ab243644 at 1/500 followed by a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101). Positive staining on human endometrial carcinoma stroma (PMID : 27446374) is observed.
The section was incubated with ab243644 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 Rabbit specific IHC polymer detection kit HRP/DAB (ab209101).

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

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab243644).

• mIHC

Supplier Data

Multiplex immunohistochemistry - Anti-PD1 antibody [EPR23119-111] - BSA and Azide free (AB270753)

This data was developed using ab243644, the same antibody clone in a different buffer formulation. Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of Human tonsil labelling PD1 with ab243644 at 1/500 dilution (1.02 µg/mL) (D), CD8 with ab178089 at 1/100 dilution (0.83 μg/ml) (C) and TIM 3 with ab242080 at 1/500 dilution (2.09 μg/ml) (B). Opal Polymer HRP Ms + Rb was used as a secondary antibody, and DAPI was used for a nuclear counter stain. Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution 2) for 20 mins.   Panel A : merged staining of anti-TIM 3 (green; Opal™690), anti-CD8 (red; Opal™520) and anti-PD1 (gray; Opal™570) on human tonsil. Panel B : anti-TIM 3 stained on membrane of a subset of immune cells. Panel C : anti-CD8 stained on membrane of a subset of T cells. Panel D : anti-PD1 stained on membrane of a subset of lymphocytes. The section was incubated in three rounds of staining : in the order of ab242080, ab243644 and ab178089 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.

• WB

Lab

Western blot - Anti-PD1 antibody [EPR23119-111] - BSA and Azide free (AB270753)

This data was developed using ab243644, the same antibody clone in a different buffer formulation.

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

This blot was developed using a higher sensitivity ECL substrate. The expression profile/ molecular weight observed is consistent with what has been described in the literature (PMID : 30487606, 31783892).

All lanes:

Western blot - Anti-PD1 antibody [EPR23119-111] (<a href='/en-us/products/primary-antibodies/pd1-antibody-epr23119-111-ab243644'>ab243644</a>) at 1/1000 dilution

Lane 1:

Untreated MOLT-4 (human lymphoblastic leukemia T lymphoblast) at 20 µg

Lane 2:

MOLT-4 treated with 500ng/ml Ionomycin and 10ng/ml Phorbol-12-myristate-13-acetate (PMA) for 24 hours at 20 µg

Secondary

All lanes:

Western blot - VeriBlot for IP Detection Reagent (HRP) (<a href='/en-us/products/reagents/veriblot-for-ip-detection-reagent-hrp-ab131366'>ab131366</a>) at 1/1000 dilution

Predicted band size: 32 kDa

Observed band size: 50-55 kDa

true

Exposure time: 48s

• WB

Lab

Western blot - Anti-PD1 antibody [EPR23119-111] - BSA and Azide free (AB270753)

This data was developed using ab243644, the same antibody clone in a different buffer formulation.

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

This blot was developed using a higher sensitivity ECL substrate. The expression profile/ molecular weight observed is consistent with what has been described in the literature (PMID : 30487606, 31783892).

All lanes:

Western blot - Anti-PD1 antibody [EPR23119-111] (<a href='/en-us/products/primary-antibodies/pd1-antibody-epr23119-111-ab243644'>ab243644</a>) at 1/1000 dilution

Lane 1:

Human tonsil tissue lysate at 20 µg

Lane 2:

Human tonsil tissue lysate treated with PNGase F at 20 µg

Secondary

All lanes:

Western blot - VeriBlot for IP Detection Reagent (HRP) (<a href='/en-us/products/reagents/veriblot-for-ip-detection-reagent-hrp-ab131366'>ab131366</a>) at 1/1000 dilution

Predicted band size: 32 kDa

Observed band size: 50-55 kDa

true

Exposure time: 48s