PD-L1 Recombinant Rabbit Monoclonal Antibody [PSH08-27]
cat.: HA722971
Product Type: Recombinant Rabbit monoclonal IgG, primary antibodies
Species reactivity: Dog
Applications: WB, IF-Cell, IHC-P
Clonality: Monoclonal
Clone number: PSH08-27
Form: Liquid
Storage condition: Store at +4℃ after thawing. Aliquot store at -20℃. Avoid repeated freeze / thaw cycles.
Storage buffer: PBS (pH7.4), 0.1% BSA, 40% Glycerol. Preservative: 0.05% Sodium Azide.
Concentration: 1ug/ul
Purification: Protein A affinity purified.
Molecular weight: Predicted band size: 33 kDa
Isotype: IgG
Immunogen: Recombinant protein within dog PD-L1 aa 1-238.
Positive control: BCMT transfected with PD-L1 cell lysate, BCMT cells transfected with PD-L1.
Subcellular location: Membrane.
Recommended Dilutions:
  WB
  IF-Cell
  IHC-P

1:2,000
1:2,500
1:5,000
Uniprot #: SwissProt: E2RKZ5 Dog
Alternative names: B7 H B7 H1 B7 homolog 1 B7-H1 B7H B7H1 CD 274 CD-274 CD274 CD274 antigen CD274 molecule MGC142294 MGC142296 OTTHUMP00000021029 PD L1 PD-L1 PD1L1_HUMAN PDCD1 ligand 1 PDCD1L1 PDCD1LG1 PDL 1 PDL1 Programmed cell death 1 ligand 1 Programmed death ligand 1 RGD1566211
Images
HA722971_1.jpg Fig1: Western blot analysis of PD-L1 on different lysates with Rabbit anti-PD-L1 antibody (HA722971) at 1/2,000 dilution.

Lane 1: BCMT cell lysate
Lane 2: BCMT transfected with PD-L1 cell lysate

Predicted band size: 33 kDa
Observed band size: 55 kDa

Exposure time: 10 seconds; ECL: K1801;

4-20% SDS-PAGE gel.

Proteins were transferred to a PVDF membrane and blocked with 5% NFDM/TBST for 1 hour at room temperature. The primary antibody (HA722971) at 1/2,000 dilution was used in 5% NFDM/TBST at 4℃ overnight. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1/50,000 dilution was used for 1 hour at room temperature.
HA722971_2.jpg Fig2: Immunocytochemistry analysis of BCMT cells transfected with PD-L1 labeling PD-L1 with Rabbit anti-PD-L1 antibody (HA722971) at 1/2,500 dilution.

Cells were fixed in 4% paraformaldehyde for 15 minutes at room temperature, permeabilized with 0.1% Triton X-100 in PBS for 15 minutes at room temperature, then blocked with 1% BSA in 10% negative goat serum for 1 hour at room temperature. Cells were then incubated with Rabbit anti-PD-L1 antibody (HA722971) at 1/2,500 dilution in 1% BSA in PBST overnight at 4 ℃. Goat Anti-Rabbit IgG H&L (iFluor™ 488, HA1121) was used as the secondary antibody at 1/1,000 dilution. PBS instead of the primary antibody was used as the secondary antibody only control. Nuclear DNA was labelled in blue with DAPI.

Beta tubulin (HA601187, red) was stained at 1/100 dilution overnight at +4℃. Goat Anti-Mouse IgG H&L (iFluor™ 594, HA1126) was used as the secondary antibody at 1/1,000 dilution.
HA722971_3.jpg Fig3: Immunohistochemical analysis of paraffin-embedded BCMT cells transfected with PD-L1 with Rabbit anti-PD-L1 antibody (HA722971) at 1/5,000 dilution.

The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (HA722971) at 1/5,000 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.
Note: All products are “FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE”.