The protein extracts were subjected to electrophoresis separation, and then the separated protein was electrotransferred to polyvinylidene fluoride membrane which was blocked with 5% bovine serum albumin (BSA) at ambient temperature for 1 hour. Then, the membrane was incubated with diluted primary antibodies: CUL3 (10450, 1/1000; Cell Signaling Technology (CST)), PD-L1 (NBP1-76769, 1/1000; Novus Biologicals), SPOP (16 750–1-AP, 1/1000; Proteintech), Flag-tag (MA1-91878, 1/1000; Sigma), GAPDH (sc-47724, 1/2000; Santa Cruz, internal reference), and HA-tag (sc-7392, 1/1000; Santa Cruz) overnight at 4°C as well as with anti-Mouse-HRP secondary antibody (7076, 1/5000; CST) or anti-Rabbit-HRP secondary antibody (7074, 1/5000; CST) for 1 hour at ambient temperature. Subsequently, the membrane was developed with ECL working solution (EMD Millipore). ImageJ analysis software was run to quantify the gray levels of each group of bands.
Pd l1 nbp1 76769
Manufactured by Novus Biologicals
PD-L1 (NBP1-76769) is an antibody that binds to the programmed cell death ligand 1 (PD-L1) protein. PD-L1 is a cell surface protein that plays a role in regulating the immune response.
Automatically generated - may contain errors
Lab products found in correlation
Flag tag (flag), by Merck Group (1 mentions)
Ha tag, by Santa Cruz Biotechnology (1 mentions)
Ecl working solution, by Merck Group (1 mentions)
Pd l1, by BioLegend (1 mentions)
Fitc mouse igg1, by BioLegend (1 mentions)
7 aad, by BioLegend (1 mentions)
P62 sqstm1, by Novus Biologicals (1 mentions)
Apc mouse igg2b, by BioLegend (1 mentions)
Dmem medium, by Thermo Fisher Scientific (1 mentions)
Lc3b nb100 2220, by Novus Biologicals (1 mentions)
Lc3a b, by Cell Signaling Technology (1 mentions)
Pd l1, by Cell Signaling Technology (1 mentions)
β actin, by Cell Signaling Technology (1 mentions)
2 protocols using pd l1 nbp1 76769
1
Western Blot Protein Detection Protocol
2
Autophagy and PD-L1 Regulation Assay
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RPMI1640 medium (72400) and DMEM medium (10564) are products from Life Technologies. 3-methyladenine (M9281), bafilomycinA1 (B1793), chloroquine (C6628), rapamycin (R0395) and phytohemagglutinin-M (PHA, L8902) are from Sigma-Aldrich. BMS 345541 (S8044) is from Selleck. The following primary antibodies were used: microtubule-associated light chain 3 (LC3B, NB100–2220, Novus Biologicals), LC3A/B (13,082, Cell Signaling), p62/SQSTM1 (H00008878-M01, Novus Biologicals), PD-L1 (NBP1–76769, Novus Biologicals), PD-L1 (59,949, Cell Signaling), PD-L1 (Spring Bio, SP142), ATG5 (12,994, Cell Signaling), ATG7 (SAB4200304, Sigma-Aldrich), β-actin (4967, Cell Signaling), CD45 (368,508, Biolegend), CD8a (301,041, Biolegend), CD4 (357,408, Biolegend), FITC Mouse IgG1(400,110, Biolegend), PD-L1 (329,708, Biolegend), APC Mouse IgG2b (300,907, Biolegend), and 7-AAD (420,404, Biolegend).
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