CD274 Antibody

• Catalog Number: orb1239770
• Category: Antibodies
• Description: CD274 Antibody
• Species/Host: Mouse
• Clonality: Monoclonal
• Clone Number: 6H10
• Tested applications: ELISA, ICC, IF, IHC-P, WB
• Predicted Reactivity: Rat
• Reactivity: Human, Mouse
• Isotype: IgG1
• Immunogen: Anti-PD-L1 antibody (orb1239770) was raised against the extracellular domain of human PD-L1.
• Concentration: 1 mg/mL
• Dilution range: WB: 0.25 - 4 μg/mL; IF: 2 μg/mL; IHC-P: 5 μg/mL; ICC: 1 μg/mL.Antibody validated: Western Blot in human and mouse samples; Immunohistochemistry, Immunocytochemistry and Immunofluorescence in human samples. All other applications and species not yet tested.
• Form/Appearance: Liquid
• Conjugation: Unconjugated
• MW: Predicted: 33kD Observed: 37kD
• Target: CD274
• UniProt ID: Q9NZQ7
• NCBI: NP_054862
• Storage: PD-L1 antibody can be stored at 4°C for three months and -20°C, stable for up to one year. As with all antibodies care should be taken to avoid repeated freeze thaw cycles. Antibodies should not be exposed to prolonged high temperatures.
• Buffer/Preservatives: PD-L1 Antibody is supplied in PBS containing 0.02% sodium azide and 50% glycerol.
• Alternative names: PD-L1 Antibody: Programmed cell death 1 ligand-1,
• Note: For research use only
• Expiration Date: 12 months from date of receipt.

Overexpression Validation of PD-L1 in 293 Cells.Loading: 15 μg of lysates per lane. Antibodies: orb1239770 (A, 0.25 μg/mL; B, 0.5 μg/mL), 1 h incubation at RT in 5% NFDM/TBST. Secondary: Goat anti-mouse IgG HRP conjugate at 1:5000 dilution.

Independent Antibody Validation (IAV) via Protein Expression Profile in Cell Lines.Loading: 15 μg of lysates per lane. Antibodies: orb1239835 (2 μg/mL), orb1239770 (2 μg/mL), and beta-actin (1 μg/mL), 1 h incubation at RT in 5% NFDM/TBST. Secondary: Goat anti-rabbit and or anti-mouse IgG HRP conjugate at 1:10000 and 1:5000 dilution, respectively.

Validation with PD-L1 siRNA Knockdown in HeLa Cells.HeLa cells were transfected with control siRNAs (lane 1) or PD-L1 siRNAs (lane 2).Loading: 10 μg of HeLa whole cell lysates per lane. Antibodies: orb1239770 (2 μg/mL) and GAPDH (orb1239765, 0.02 μg/mL), 1 h incubation at RT in 5% NFDM/TBST. Secondary: Goat anti-mouse IgG HRP conjugate at 1:5000 dilution.

Western Blot Validation of PD-L1 in Raji Cells.Loading: Lysates/proteins at 15 μg per lane. Antibodies: orb1239770 (4 μg/mL). 1 h incubation at RT in 5% NFDM/TBST. Secondary: Goat anti-mouse IgG HRP conjugate at 1:10000 dilution.

Immunofluorescence Validation of PD-L1 in Transfected 293 Cells.Immunofluorescent analysis of 4% paraformaldehyde-fixed PD-L1 transfected 293 cells labeling PD-L1 with orb1239770 at 2 μg/mL, followed by goat anti-mouse IgG secondary antibody at 1/500 dilution (red) and DAPI staining (blue).

Immunofluorescence Validation of PD-L1 in Human Stomach Carcinoma Tissue. Immunofluorescent analysis of 4% paraformaldehyde-fixed human stomach carcinoma tissue labeling PD-L1 with orb1239770 at 2 μg/mL, followed by goat anti-mouse IgG secondary antibody at 1/500 dilution (red) and DAPI staining (blue).

Immunofluorescence Validation of PD-L1 in Human Tonsil Tissue.Immunofluorescent analysis of 4% paraformaldehyde-fixed human tonsil tissue labeling PD-L1 with orb1239770 at 2 μg/mL, followed by goat anti-mouse IgG secondary antibody at 1/500 dilution (red) and DAPI staining (blue).

Immunohistochemistry Validation of PD-L1 in Human Stomach Carcinoma Tissue.Immunohistochemical analysis of paraffin-embedded human stomach carcinoma tissue using anti-PD-L1 antibody (orb1239770) at 5 μg/ml. Tissue was fixed with formaldehyde and blocked with 10% serum for 1 h at RT; antigen retrieval was by heat mediation with a citrate buffer (pH6). Samples were incubated with primary antibody overnight at 4°C. A goat anti-mouse IgG H&L (HRP) at 1/250 was used as secondary. Counter stained with Hematoxylin. Lower left: Use mouse IgG antibody at 1 μg/ml as control.

Immunohistochemistry Validation of PD-L1 in Human Tonsil Carcinoma Tissue.Immunohistochemical analysis of paraffin-embedded human tonsil tissue using anti-PD-L1 antibody (orb1239770) at 5 μg/ml. Tissue was fixed with formaldehyde and blocked with 10% serum for 1 h at RT; antigen retrieval was by heat mediation with a citrate buffer (pH6). Samples were incubated with primary antibody overnight at 4°C. A goat anti-mouse IgG H&L (HRP) at 1/250 was used as secondary. Counter stained with Hematoxylinl. Lower left: Use mouse IgG antibody at 1 μg/ml as control.

Immunocytochemistry Validation of PD-L1 in PD-L1 Overexpressed 293 Cells. Immunocytochemical analysis of PD-L1 transfected 293 cells using anti- PD-L1 antibody (orb1239770) at 1 μg/mL. Cells was fixed with formaldehyde and blocked with 10% serum for 1 h at RT; antigen retrieval was by heat mediation with a citrate buffer (pH6). Samples were incubated with primary antibody overnight at 4° C. A goat anti-rabbit IgG H&L (HRP) at 1/250 was used as secondary. Counter stained with Hematoxylin. Lower left: Use mouse IgG antibody at 1 μg/mL as control.