Western blots were performed with antibodies against of GFP (Abcam no. ab290, 1:2,000), MYC (Cell Signaling no. 5605, 1:1,000), β-actin (Sigma no. A5316, 1:10,000), ARG1 (Sigma no. HPA024006, 0.4 μg ml–1 for western blot), RASG12D (Cell Signaling no. 14429, 1:1,000), p-eIF2α Ser51 (Cell Signaling no. 3597, 1:500), eIF2α (Cell Signaling no. 9722, 1:1,000), p-eIF4E Ser209 (Cell Signaling no. 9721, 1:1,000), eIF4E (BD no. 610270, 1:1,000), GAPDH (Cell Signaling no. 2118, 1:1,000), p-ERK1/2 (Cell Signaling no. 4370, 1:1,000), SNAIL (Proteintech, 13099–1-AP, 1:500), CDC20 (Proteintech, 10252–1-AP, 1:1,000), PLK1 (Proteintech, 10305–1-AP, 1:1,000), ADAM8 (Proteintech, 23778–1-AP, 1:1,000), RPS2 (Abcam, ab58341, 1:1,000), HA-HRP (Cell Signaling no. 2999, 1:1,000), HSP90 (Cell Signaling, no. 4877, 1:1,000), and PD-L1 (Cell Signaling, no. 13684, 1:1,000). Blots are developed using ChemiDoc Imaging Systems (BioRad).
Adam8
Manufactured by Proteintech
Sourced in United States, China
ADAM8 is a laboratory equipment product manufactured by Proteintech. It is a metallic tool used for specific applications in research and scientific experiments. The core function of ADAM8 is to facilitate precise measurements and sample handling during laboratory procedures.
Automatically generated - may contain errors
Lab products found in correlation
Peif4eser209, by Cell Signaling Technology (1 mentions)
Eif2α, by Cell Signaling Technology (1 mentions)
Chemidoc imaging system, by Bio-Rad (1 mentions)
P erk1 2, by Cell Signaling Technology (1 mentions)
P eif2α ser51, by Cell Signaling Technology (1 mentions)
Eif4e, by BD (1 mentions)
Rasg12d, by Cell Signaling Technology (1 mentions)
Cdc20, by Proteintech (1 mentions)
Snail, by Proteintech (1 mentions)
Hsp90, by Cell Signaling Technology (1 mentions)
Pd l1, by Cell Signaling Technology (1 mentions)
Gapdh, by Cell Signaling Technology (1 mentions)
β actin, by Merck Group (1 mentions)
Hb egf, by ABclonal (1 mentions)
Gapdh, by Enogene (1 mentions)
P akt, by Proteintech (1 mentions)
Bca kit, by Beyotime (1 mentions)
Tgf β1, by ABclonal (1 mentions)
Ripa lysis, by Beyotime (1 mentions)
Ve cadherin, by ABclonal (1 mentions)
4 protocols using adam8
1
Western Blot Antibody Validation Protocol
2
Protein Extraction and Western Blot Analysis
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Total protein was extracted with RIPA buffer, and 20–50 μg samples were loaded after measuring their concentration using a BCA kit and separated by 6%, 8% or 12% sodium dodecyl sulfate-polyacrylamide gel electrophoresis. The separated proteins were transferred onto polyvinylidene fluoride membrane blocked with 5% fat-free milk for 2 h at room temperature and incubated in primary antibodies against ADAM8 (1:1000, Proteintech), HB-EGF (1:1000, Abclonal Technology), EGFR (1:1000, Proteintech), p-EGFR (1:1000, CST, Beverly, MA, USA), ERK (1:1000, CST), p-ERK (1:1000, CST), AKT (1:1000, Proteintech), p-AKT (1:1000, Proteintech), CCL2 (1:1000, Proteintech), and GAPDH (1:2000, Enogene, New York, NY, USA) at 4 °C overnight. Membranes were washed and incubated in horseradish peroxidase (HRP)-conjugated secondary antibodies for 1 h at room temperature. An enhanced chemiluminescence system (NCM Biotech, Suzhou, China) was used to detect the protein bands.
3
Protein Extraction and Western Blot Analysis
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The protein of tissues and cells were extracted via RIPA lysis (Beyotime). The concentration of protein was measured using BCA kits (Beyotime). Following, the protein was separated by sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE), and were transferred onto polyvinylidene fluoride (PVDF) membranes. After blocking, the membranes were incubated with the primary antibodies (ADAM8, 1:1,000, Proteintech; α-SMA, 1:1,000, Affinity; FSP1, 1:1,000, Affinity; VE-cadherin, 1:1,000, ABclonal, Wuhan, China; TGF-β1, 1:1,000, ABclonal; Smad2, 1:1,000, Affinity; p-Smad2, 1:1,000, Affinity; Smad3, 1:1,000, ABclonal; p-Smad3, 1:1,000, ABclonal) at 4°C overnight. The membrane was washed using PBS following incubation with corresponding secondary antibodies. The image was analyzed by Gel-Pro Analyzer (Media Cybernetics, Bethesda, MD, USA).
4
Immunohistochemical Analysis of Immune Markers
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Immunohistochemistry staining was performed as described in our previous publication [25 (link)]. Primary antibodies included ADAM8 (1:100, Proteintech), IBA-1 (1:500, Abcam), CD206 (1:1000, Abcam). The percentage of positive cells or the percentage of positive areas was calculated in five randomly selected fields using ImageJ software (ImageJ 1.8.0, NIH, Bethesda, MD, USA) respectively.
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