Anti-PD1 抗体 [NAT105]

• IHC-Fr

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Immunohistochemistry (Frozen sections) - Anti-PD1 antibody [NAT105] (AB52587)

Immunohistochemical analysis of permeabilized frozen Human tonsil tissue labeling PD1 with ab316965 at 1/10000 dilution, followed by LeicaDS9800 (Bond™ PolymerRefine Detection). Counterstained with hematoxylin. The immunostaining was performed on a Leica Biosystems BOND® RX instrument.

• ICC/IF

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Immunocytochemistry/ Immunofluorescence - Anti-PD1 antibody [NAT105] (AB52587)

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.

• IHC-Fr

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Immunohistochemistry (Frozen sections) - Anti-PD1 antibody [NAT105] (AB52587)

Immunohistochemical analysis of permeabilized frozen Human cardiac muslce tissue labeling PD1 with ab316965 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.

• IHC-P

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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-PD1 antibody [NAT105] (AB52587)

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.

• Flow Cyt

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Flow Cytometry - Anti-PD1 antibody [NAT105] (AB52587)

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.

• IHC-P

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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-PD1 antibody [NAT105] (AB52587)

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.

• WB

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Western blot - Anti-PD1 antibody [NAT105] (AB52587)

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

All lanes:

Western blot - Anti-PD1 antibody [NAT105] (ab52587) 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

false