Western blot analysis was applied to detect protein levels. Briefly, proteins were extracted from the injured cerebral cortex with a protein extraction kit purchased from Beyotime Bio Inc. Then, 30 µg protein of each sample was separated by SDS-PAGE and then transferred to polyvinylidene fluoride (PVDF) membranes by wet transfer. The PVDF membranes were blocked by 5% BSA or 5% milk and then incubated at 4 °C overnight with primary antibodies against ENT1 (1:1000, ab223851, Abcam), NLRP3 (1:1000, ab4207, Abcam), Bcl2 (1:1000, AF6139, affinity), β-actin (1:3000, affinity), Bax (1:1000, AF0120, affinity), apoptosis associated speck like protein containing CARD (ASC) (1:1000, DF6304, affinity), caspase-1 (1:1000, ab1871, Millipore) and IL-1β (1:1000, ab9722, Abcam). After washing with tris buffered saline tween, the PVDF membranes were incubated with secondary antibodies at room temperature for 1 h. Enhanced chemiluminescence Plus Chemiluminescence Reagent Kit was used to detect positive protein bands, results were quantified by Image J software.
Af6139
Manufactured by Abcam
Sourced in China
AF6139 is a primary antibody that recognizes the protein Human Apolipoprotein E. The product is suitable for use in various immunoassay applications.
Automatically generated - may contain errors
Lab products found in correlation
Ab4207, by Abcam (1 mentions)
Ab1871, by Merck Group (1 mentions)
Ab9722, by Abcam (1 mentions)
Protein extraction kit, by Beyotime (1 mentions)
Ab55831, by Abcam (1 mentions)
Clarity western ecl substrate, by Bio-Rad (1 mentions)
Bm0627, by Boster Bio (1 mentions)
Ab80508, by Wuhan Servicebio Technology (1 mentions)
Polyvinylidene difluoride membrane, by Merck Group (1 mentions)
V900933, by Merck Group (1 mentions)
Af0120, by Affinity Biosciences (1 mentions)
Bovine serum albumin (bsa), by Merck Group (1 mentions)
2 protocols using af6139
1
Cerebral Cortex Protein Analysis
2
Protein Expression Analysis in Seminal Vesicles
Check if the same lab product or an alternative is used in the 5 most similar protocols
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The cryopreserved seminal vesicles were ground and incubated with lysis buffer to extract the protein. 40 µg protein lysate of each sample was electrophoresed in sodium dodecyl sulfate/polyacrylamide (SDS-PAGE) gel and transferred to a polyvinylidene difluoride membrane (Millipore, Burlington, MA, USA). Then the membrane was blocked with 5% bovine serum albumin (Sigma-Aldrich; V900933) for 1 h and incubated with the primary antibodies at 4 °C overnight, including antibodies of Bax (1:500; Affinity, AF0120, Wuhan, China), Bcl-2 (1:500; Affinity, AF6139), NOX1 (1:500; Abcam, ab55831), NOX2 (1:500; Abcam, ab80508), NOX4 (1:500; Servicebio, GB11347, Wuhan, China),and β-actin (1:500; BM0627; Boster, Wuhan, China). After incubation with horseradish peroxidase-conjugated secondary antibodies (1:1000; 7074, 7076; CST, Danvers, MA, USA), the membranes were developed using Bio-Rad Clarity Western ECL Substrate (1705061; Bio-Rad Laboratories, Hercules, CA, USA). Protein bands were analyzed with Image J software (National Institutes of Health, Bethesda, MD, USA).
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