Total protein was extracted from tissues by RIPA lysis buffer (Beyotime), 10 μL PMSF and 10 μL phosphatase inhibitor. SDS-PAGE gel electrophoresis was utilized to separate the extracted protein. The protein was transferred to polyvinylidene fluoride membrane. After blocking with 50 g/L BAS for 1 h, the membrane was incubated with primary antibodies against HTR2B (1/3000; ab227722; Abcam, Cambridge, MA, United States), SLC5A3 (1/1000; ab113245), GAPDH (1/3000; ab8245), TNF-α (1/1000; ab215188), IL-1β (1/1000; ab200478), TGF-β (9ab208156) and IL-4 (1/1000; ab34277) at 4°C overnight. After washing the membrane with TBST, the membrane was incubated with secondary antibody (1/5000; ab7097) for 1.5 h. The ultra-sensitive ECL chemiluminescence kit was used to configure the luminescence buffer, and the band was developed.
Ab34277
Manufactured by Abcam
Sourced in United States
Ab34277 is a lab equipment product. It is a generic laboratory product used for various research and experimental purposes. No further details are available about its specific core function or intended use.
Automatically generated - may contain errors
Lab products found in correlation
Ab113245, by Abcam (1 mentions)
Ab215188, by Abcam (1 mentions)
Ab208156, by Abcam (1 mentions)
Ab200478, by Abcam (1 mentions)
Ripa lysis buffer, by Beyotime (1 mentions)
Ab8245, by Abcam (1 mentions)
Ab109461, by Abcam (1 mentions)
Ab181603, by Abcam (1 mentions)
Ab231172, by Abcam (1 mentions)
Ab263947, by Abcam (1 mentions)
Ab207086, by Abcam (1 mentions)
Ab133543, by Abcam (1 mentions)
Ab200738, by Abcam (1 mentions)
Ab134157, by Abcam (1 mentions)
Ab76315, by Abcam (1 mentions)
2 protocols using ab34277
1
Protein Extraction and Western Blot Analysis
2
Protein Expression Analysis in Cellular Aging
Check if the same lab product or an alternative is used in the 5 most similar protocols
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After removing the culture medium, 500 μl protein lysate is added to each well for lysis for 30 minutes, centrifuged at 4° C, 1500 r/min for 5 minutes. And the supernatant is taken to detect the protein concentration AGING of the sample. Transfer the protein to PVDF membrane through Gels preparation, samples loading, electrophoresis and wet transfer and close the blocking solution for 1 hour. Sequentially, primary antibody SHP2 (Abcam, ab187040, 1:5000), Ki-67 (Abcam, ab231172, 1:5000), p-STAT1 (Abcam, ab109461, 1:1000), p-STAT3 (Abcam, ab76315, 1:1000), p-STAT5 (Abcam, ab278764, 1:1000), p-STAT6 (Abcam, ab263947, 1:1000), IL-4 (Abcam, ab34277, 1:1000), IL-10 (Abcam, ab52909, 1:1000), Cathepsin-L (Abcam, ab200738, 1:1000), Cathepsin-S (Abcam, ab134157, 1:1000), Cathepsin-K (Abcam, ab207086, 1:1000), Arginase-1 (Abcam, ab133543, 1:1000) and GAPDH (Abcam, ab181603, 1:10000) are added and the secondary antibody is incubated for 2 hours at room temperature. Exposure imaging is performed and Quantity One V4 software is used for analysis.
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