Anti-PD1 antibody [NAT105] - BSA and Azide free

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-PD1 antibody [NAT105] - BSA and Azide free (AB201811)

Immunohistochemical analysis of paraffin-embedded Human cardiac muscle tissue labeling PD1 with ab52587 at 1/100 dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection). No staining on human cardiac muscle. The immunostaining was performed on a Leica Biosystems BOND® RX instrument. Counterstained with Hematoxylin. Heat mediated antigen retrieval was performed with Citrate buffer (pH 6.0, Epitope Retrieval Solution 1) for 20 mins.

This data was developed using the same antibody clone in a different buffer formulation containing BSA and sodium azide (ab52587).

• IHC-Fr

Supplier Data

Immunohistochemistry (Frozen sections) - Anti-PD1 antibody [NAT105] - BSA and Azide free (AB201811)

Immunohistochemical analysis of permeabilized frozen Human tonsil tissue labeling PD1 with ab52587 at 1/10000 dilution, followed by LeicaDS9800 (Bond™ PolymerRefine Detection). Counterstained with hematoxylin. The immunostaining was performed on a Leica Biosystems BOND® RX instrument.

This data was developed using the same antibody clone in a different buffer formulation containing BSA and sodium azide (ab52587).

• mIHC

Collaborator

Multiplex immunohistochemistry - Anti-PD1 antibody [NAT105] - BSA and Azide free (AB201811)

This data was developed using the same antibody clone in a different buffer formulation containing BSA and sodium azide (ab52587).

10-color fluorescence multiplex immunohistochemical analysis of human lung cancer tissue (formalin-fixed paraffin-embedded section).

Merged staining of anti-FOXP3 (ab215206; Cyan; TG540N), anti-PD1 (ab52587; Red; TG700N), anti-CD163 (ab182422; Brown; TG650N), anti-HLA-DR (ab92511; Yellow; TG570N), anti-CD4 (ab133616; Violet; TG620N), anti-CD8 alpha (ab101500; Purple; TG540S), anti-CD20 (ab9475; Grey; TG660S), anti-CD68 (ab192847; Green; TG520N), anti-Cytokeratin 19 (ab52625; Light blue; TG440N). TG470SN (dark blue) was used as a nuclear counter stain. The inset image shows the separate CD68 signal.

The section was incubated in nine rounds of staining; in the order of ab215206 (1/100 dilution), ab52587 (1/200 dilution), ab182422 (1/300 dilution), ab92511 (1/200 dilution), ab133616 (1/600 dilution), ab101500 (1/300 dilution), ab9475 (1/100 dilution), ab192847 (1/300 dilution), ab52625 (1/400 dilution); each using a separate fluorescent tyramide signal amplification system.

Sodium citrate antigen retrieval (pH6.0) was used in between rounds of tyramide signal amplification to remove the antibody from the previous round, to avoid any cross-reactivity.

Image acquisition was performed with TissueFAXS Spectra (TissueGnostics).

This image is courtesy of TissueGnostics Asia Pacific Limited

• IHC-Fr

Supplier Data

Immunohistochemistry (Frozen sections) - Anti-PD1 antibody [NAT105] - BSA and Azide free (AB201811)

Immunohistochemical analysis of permeabilized frozen Human cardiac muslce tissue labeling PD1 with ab52587 at 1/10000 dilution, followed by LeicaDS9800 (Bond™ PolymerRefine Detection). No staining on human cardiac muscle. Counterstained with hematoxylin. The immunostaining was performed on a Leica Biosystems BOND® RX instrument.

This data was developed using the same antibody clone in a different buffer formulation containing BSA and sodium azide (ab52587).

• ICC/IF

Supplier Data

Immunocytochemistry/ Immunofluorescence - Anti-PD1 antibody [NAT105] - BSA and Azide free (AB201811)

Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% TritonX-100 permeabilized MOLT-4 cells labelling PD1 with ab52587 at 1/100 dilution, followed by Goat Anti-Mouse IgG H&L (Alexa Fluor^® 488) preadsorbed (ab150117) at 1/1000 dilution at RT for 45 min. Recombinant Alexa Fluor^® 594 Anti-beta Tubulin antibody [EPR16774] (ab206369) was used as a counterstain at 1/50 dilution and was co-incubated with ab52587 overnight at 4° C. Nucleus were visualized using DAPI. Confocal image showing membranous staining in MOLT-4 cells treated with 500 ng/ml Ionomycin and 10 ng/ml Phorbol-12-myristate-13-acetate (PMA) for 24 hours, and no staining in MOLT-4 cells.

This data was developed using the same antibody clone in a different buffer formulation containing BSA and sodium azide (ab52587).

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-PD1 antibody [NAT105] - BSA and Azide free (AB201811)

Immunohistochemical analysis of paraffin-embedded Human tonsil tissue labeling PD1 with ab52587 at 1/100 dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection). The immunostaining was performed on a Leica Biosystems BOND® RX instrument. Counterstained with Hematoxylin. Heat mediated antigen retrieval was performed with Citrate buffer (pH 6.0, Epitope Retrieval Solution 1) for 20 mins.

This data was developed using the same antibody clone in a different buffer formulation containing BSA and sodium azide (ab52587).

• Flow Cyt

Supplier Data

Flow Cytometry - Anti-PD1 antibody [NAT105] - BSA and Azide free (AB201811)

Cells were fixed with 100µl 4% PFA for 10 min at RT, then permeabilized with 100µl 90% methanol for 30 min (-20° C) followed by blocking with 10% goat serum for 1h at room temperature. Incubate ab52587 for 30 min at room temperature (RT) and then incubate Goat anti-Mouse IgG (Alexa Fluor^® 488, ab150113) for 30 min at room temperature. Positive staining on MOLT-4 treated with 10ng/ml PMA and 500ng/ml Ionomycin for 24h, but weak staining on untreated MOLT-4.

This data was developed using the same antibody clone in a different buffer formulation containing BSA and sodium azide (ab52587).

• WB

Supplier Data

Western blot - Anti-PD1 antibody [NAT105] - BSA and Azide free (AB201811)

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

This data was developed using the same antibody clone in a different buffer formulation containing BSA and sodium azide (ab52587).

All lanes:

Western blot - Anti-PD1 antibody [NAT105] (<a href='/en-us/products/primary-antibodies/pd1-antibody-nat105-ab52587'>ab52587</a>) at 1/1000 dilution

Lane 1:

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

Lane 2:

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

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