Anti-PD1 抗体 [EPR4877(2)] - BSA and Azide free (ab186928)

Immunohistochemical analysis of formalin fixed paraffin embedded human tonsil labelling PD-L1 with ab186928 at a concentration of 5µg/ml. The immunostaining was performed on a Ventana DISCOVERY ULTRA (Roche Tissue Diagnostics) instrument with an OptiView DAB IHC Detection Kit. Heat mediated antigen retrieval was conducted for 32min with ULTRA cell conditioning solution (CC1 pH8.5). ab186928 anti PD-L1 antibody was incubated at 37°C for 16min. Sections were counterstained is with Hematoxylin II. Image inset shows absence of staining in secondary antibody only control.

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human tonsil tissue labelling PD1 with purified ab137132 at 1/500. Heat mediated antigen retrieval was performed using Tris/EDTA buffer pH 9. ab97051, a HRP-conjugated goat anti-rabbit IgG (H+L) was used as the secondary antibody (1/500). Negative control using PBS instead of primary antibody. Counterstained with hematoxylin.

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab137132).

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human tonsil tissue labelling PD1 with ab137132 at a dilution of 1/500. Heat mediated antigen retrieval was performed using EDTA antigen retrieval solution, and microwave treatment for 20 min at 20% power. Anti-Rabbit HRP polymer was used as secondary antibody. Opal tyramide amplification was performed using Opal 540 fluorophore. Counterstained with DAPI stain.

Image scanned with Vectra 3.0 and analyzed via Inform software.

This image was courteously provided by Dr. Houssein Abdul Sater, Georgia Cancer Center.

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab137132).

Immunohistochemical (Formalin/PFA-fixed paraffin-embedded sections) analysis of human tonsil tissue labeling PD1 with ab137132 at a dilution of 1/250. Anti-Rabbit HRP polymer was used as the secondary detection system. Heat-mediated antigen retrieval was performed using EDTA based pH 9.0 buffer.

Immunohistochemical (Formalin/PFA-fixed paraffin-embedded sections) analysis of human tonsil tissue labeling PD1 with ab137132 at a dilution of 1/250. Anti-Rabbit HRP polymer was used as the secondary detection system. Heat-mediated antigen retrieval was performed using citrate based pH 6.0 buffer.

Clone EPR4877(2) (ab186928) has been successfully conjugated by Abcam. This image was generated using Anti-PD1 antibody [EPR4877(2)] (Alexa Fluor® 647). Please refer to ab201825 for protocol details.

IHC image of PD1 staining in a section of formalin-fixed paraffin-embedded normal human tonsil*.

The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH6) in a Dako Pascal pressure cooker using the standard factory-set regime. Non-specific protein-protein interactions were then blocked in TBS containing 0.025% (v/v) Triton X-100, 0.3M (w/v) glycine and 1% (w/v) BSA for 1h at room temperature.  The section was then incubated overnight at +4°C in TBS containing 0.025% (v/v) Triton X-100 and 1% (w/v) BSA with ab201825 at 1/100 dilution (shown in red) and counterstained using ab195887, Mouse monoclonal to alpha Tubulin (Alexa Fluor^® 488), at 1/250 dilution (shown in green). Nuclear DNA was labelled with DAPI (shown in blue). The section was then mounted using Fluoromount^®.

Image was taken with a confocal microscope (Leica-Microsystems, TCS SP8).

For other IHC staining systems (automated and non-automated), customers should optimize variable parameters such as antigen retrieval conditions, antibody concentrations and incubation times.

*Tissue obtained from the Human Research Tissue Bank, supported by the NIHR Cambridge Biomedical Research Centre.

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human T-cell lymphoma tissue labelling PD1 with unpurified ab137132 at a dilution of 1/250.

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab137132).

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human tonsil tissue labelling PD1 with unpurified ab137132 at a dilution of 1/250.

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab137132).

Immunohistochemical analysis using ab137132 showing negative staining in normal human brain tissue.

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab137132).

Immunohistochemical analysis using ab137132 showing negative staining in human skeletal muscle tissue.

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab137132).

Immunohistochemistry (Paraffin-embedded sections) analysis of Human angioimmunoblastic T-cell lymphoma tissue labeling PD1 with ab186928 at 1/1000. Heat mediated antigen retrieval was perfomed using Tris/EDTA buffer pH 9. Anti-rabbit IgG H&L (HRP) (ab97051) was used as the secondary antibody at 1/500. Counterstained with hematoxylin.

Immunohistochemistry (Paraffin-embedded sections) analysis of Human tonsil tissue labeling PD1 with ab186928 at 1/1000. Heat mediated antigen retrieval was perfomed using Tris/EDTA buffer pH 9. Anti-rabbit IgG H&L (HRP) (ab97051) was used as the secondary antibody at 1/500. Counterstained with hematoxylin.

Tissue Microarrays stained for " Anti-PD1 antibody [EPR4877(2)]” using " ab137132" 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 Bond™ Epitope Retrieval Solution 2 (pH 9.0) for 20 minutes. The sections were incubated with ab137132 for 30 mins at room temperature followed by a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101). The immunostaining was performed on a Leica Biosystems BOND® RX instrument.