Cells were lysed with RIPA lysis buffer (20 mM Tris-HCl, 37 mM NaCl2, 2 mM EDTA, 1% Triton-X, 10% glycerol, 0.1% SDS, and 0.5% sodium deoxycholate) with protease and phosphatase inhibitors (Roche). Protein samples were quantified (Pierce BCA Protein Assay Kit, Thermo Fisher Scientific), subjected to SDS-PAGE and transferred to PVDF membranes. Membranes were incubated with primary antibodies, including RXRA (1:1000, Proteintech, Cat. 21218–1-AP) and ALOX5 (1:1000, Abclonal, Cat. A2877), and subsequent secondary antibody: HRP-conjugated Affinipure Goat Anti-Rabbit IgG(H+L) (1:3000, Proteintech, Cat. SA00001–2).
Alox5
Manufactured by ABclonal
Sourced in China
ALOX5 is a laboratory equipment product manufactured by ABclonal. It is a tool used for the detection and analysis of the 5-lipoxygenase enzyme, which plays a key role in the biosynthesis of leukotrienes. The product provides researchers with the necessary components to study the activity and function of this important enzyme in various biological systems.
Automatically generated - may contain errors
Lab products found in correlation
Hrp conjugated affinipure goat anti rabbit igg h l, by Proteintech (1 mentions)
Abcc1, by Abcam (1 mentions)
Acsl4, by ABclonal (1 mentions)
Glyceraldehyde 3, by Proteintech (1 mentions)
Odyssey, by LI COR (1 mentions)
Pvdf membrane, by Merck Group (1 mentions)
Ripa lysis buffer, by Beyotime (1 mentions)
Bca protein assay kit, by Beyotime (1 mentions)
Gapdh, by Proteintech (1 mentions)
2 protocols using alox5
1
Quantification and Western Blot Analysis
2
Protein Analysis of Skin Tissue
Check if the same lab product or an alternative is used in the 5 most similar protocols
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Lysis of skin tissue was performed in RIPA lysis buffer (Beyotime, China). A bicinchoninic acid (BCA) Protein Assay Kit (Beyotime, China) was used to measure protein concentration. A sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) gradient of 8% to 12% was applied to detect total proteins, followed by polyvinylidene fluoride (PVDF) membrane (Millipore, USA) transfer to separate the bands. Skim milk (5%) was used to block non-specific binding for 1 hour at room temperature. Primary antibodies were then incubated with acyl-CoA synthetase long-chain family member 4 (ACSL4; 1꞉1 000; Abclonal, China), arachidonate 5-lipoxygenase (ALOX5; 1꞉1 000; Abclonal, China), zinc finger E-box-binding homeobox 1 (ZEB1; 1꞉1 000; Abclonal, China), ChaC glutathione specific gamma-glutamylcyclotransferase 1 (CHAC1; 1꞉1 000; Abcam, UK), ATP binding cassette subfamily C member 1 (ABCC1; 1꞉1 000; Abcam, UK), acetyl-CoA carboxylase alpha (ACACA; 1꞉1 000; Abclonal, China), and cytochrome p450 oxidoreductase (POR; 1꞉1 000; Abcam, UK), and glyceraldehyde-3-phosphate dehydrogenase (GAPDH; 1꞉3 000; Proteintech Group, USA). As a secondary antibody, anti-HRP-mouse IgG or anti-HRP-rabbit IgG (1꞉3 000; Abcam, UK) was used. By using Odyssey Li-Cor (LI-COR, USA), blots were revealed and quantified.
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