Total protein was extracted using RIPA buffer containing proteinase inhibitor (Best Biological, Jiangsu, China). Western blot was conducted as previously described [12 (link)]. The primary antibodies for P-ERK1, ERK1, NF-κB1, RELA, P-mTOR, mTOR, VEGFA, PD-L1, P-P13K, P13K, P-AKT, AKT, TNF-α, P-EGFR, EGFR and HIF-1α were purchased from Bioss (Beijing, China). Total protein level was normalized to β-actin.
Pd l1
Manufactured by Bioss Antibodies
Sourced in China
The PD-L1 is a laboratory instrument used to detect and quantify the expression of the Programmed Death-Ligand 1 (PD-L1) protein. PD-L1 is a key immune checkpoint molecule that plays a critical role in regulating the immune response. The PD-L1 instrument provides researchers with a reliable and accurate method to measure PD-L1 expression levels in various biological samples, which is essential for understanding immune system function and developing targeted therapies.
Automatically generated - may contain errors
Lab products found in correlation
Tnf α, by Bioss Antibodies (1 mentions)
Hif 1α, by Bioss Antibodies (1 mentions)
P akt, by Bioss Antibodies (1 mentions)
P mtor, by Bioss Antibodies (1 mentions)
Vegfa, by Bioss Antibodies (1 mentions)
Tcs sp8, by Leica camera (1 mentions)
35 mm glass bottom culture dishes, by MatTek (1 mentions)
Ecl plus reagent, by Beyotime (1 mentions)
P nf κb, by Bioss Antibodies (1 mentions)
Nf κb, by Santa Cruz Biotechnology (1 mentions)
P erk1 2, by Bioss Antibodies (1 mentions)
Pvdf membrane, by Bio-Rad (1 mentions)
Ripa buffer, by Beyotime (1 mentions)
Ifn β, by Bioss Antibodies (1 mentions)
Bs 1103r, by Bioss Antibodies (1 mentions)
4 protocols using pd l1
1
Comprehensive Protein Analysis via Western Blot
2
Evaluating Nanoparticle Cellular Uptake
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To evaluate the cellular uptake of the nanoparticles, 4T1 cells were seeded onto 35 mm glass-bottom culture dishes (MatTek, USA) at a density of 1 × 105 cells/mL overnight. Then, the cells were incubated with SPIO-aPD-L1-Cy5.5 NPs (5 μg/ml, 1 mL) at pH 7.4 for 4 h, and SPIO-IgG-Cy5.5 NPs were used as controls. To further validate the active targeting of the nanoparticles, a blocking group was made, which was designed to add antibody PD-L1 (Bioss, China) before adding SPIO-aPD-L1-Cy5.5. After the incubation, the cells were then washed three times in PBS, and DAPI was used to stain the nuclei for 10 minutes. Fluorescence images were acquired by confocal laser scanning microscopy (TCS SP8, Leica, Germany).
3
Protein Extraction and Western Blot Analysis
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Total proteins were extracted using RIPA buffer (Beyotime, Shanghai, China) according to the manufacturer’s instructions. The protein lysate was subjected to electrophoresis and transferred onto a PVDF membrane (Bio-Rad, CA, United States). The following primary antibodies sourced from CST (Shanghai, China): p-MEK (Ser217/221; 1:1000; 9154), MEK (1:1000; 8727), p-ERK1/2 (Thr202/Tyr204; 1:2000; 4370), and ERK (1:1000; 4695), PD-1 (1:1000; 84651), PD-L1 (1:1000; 60475), and p-NF-κB (1:1000; bs-0982R) from Bioss (Beijing, China), NF-κB (1:1000; sc-8008) from Santa Cruz Biotechnology (Texas, United States) were used to incubate the membrane, respectively. The proteins were detected using ECL Plus Reagent (Beyotime) and quantified using Image Lab software.
4
Comprehensive Immunohistochemical Profiling
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Pre- and post-treatment tissue specimens were stained for STING (Cell signaling technology, #13647S, 1:100), PD-L1 (Bioss, bs-1103R, 1:300), IFN-β (Bioss, bs-23731R, 1:300), CD3 (Proteintech, 17617-1-AP, 1:100), CD8 (Proteintech, 66868-1-Ig, 1:1000). 5-um-thick tissue sections were deparaffinized in xylene, passed through graded alcohols, and antigen repaired with citrate buffer (pH=7.6) in a steam pressure cooker. The sections were blocked with 5% BSA 1H and incubated with primary antibodies at 4°C overnight, and then washed in 50 mM Tris-HCl, pH 7.4 and incubated with horseradish peroxidase-conjugated secondary antibodies. Immunoperoxidase staining was performed with DAB system. Slides were counterstained with haematoxylin. Immunofluorescence-stained sections were added dropwise with DAPI, incubated for 10 minutes and then rinsed 3 times with PBS. Immunohistochemical sections were dehydrated in in graded alcohol and xylene, and sealed with neutral gum. Immunostaining sections were sealed with glycerin.
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