The cell lysate was collected using the lysis buffer (Beyotime Biotechnology, Shanghai, China). The whole proteins were scattered through SDS–PAGE on 10% gel and were transferred to polyvinylidene difluoride membranes (Provider). Membranes were blocked using 5% fat-free milk in PBS-Tween 20 and incubated with primary antibodies at 4 °C overnight. The primary antibodies were anti-RBP1 (1:1000; Proteintech, cat. no. 22683-1-AP), anti-ATG5 (1:500; Proteintech, cat. no. 10181-2-AP), anti-Beclin 1 (1:1000; Proteintech, cat. no. 11306-1-AP), anti-LC3B (1:1000; Cell Signaling Technology, cat. no. 2775), and anti-β-actin (1:2000; Proteintech, cat. no. 20536-1-AP). Subsequently, the membranes were incubated with secondary antibody Goat Anti-Rabbit IgG (1:10,000; Proteintech, cat. no. 10285-1-AP) for 1 h at room temperature.
Anti rbp1
Manufactured by Proteintech
Sourced in United States
Anti-RBP1 is a polyclonal antibody that specifically recognizes the Retinol Binding Protein 1 (RBP1) protein. RBP1 is a cellular retinol-binding protein involved in retinol (vitamin A) transport and metabolism. The antibody can be used for the detection of RBP1 in various experimental applications.
Automatically generated - may contain errors
Lab products found in correlation
Anti rabbit igg, by Proteintech (1 mentions)
Anti atg5, by Proteintech (1 mentions)
Anti beclin 1, by Proteintech (1 mentions)
Anti lc3b, by Cell Signaling Technology (1 mentions)
Anti β actin, by Proteintech (1 mentions)
Lysis buffer, by Beyotime (1 mentions)
Enhanced chemiluminescence reagent, by Santa Cruz Biotechnology (1 mentions)
Anti hsp90, by Proteintech (1 mentions)
Anti cd9, by Proteintech (1 mentions)
Cemip, by Proteintech (1 mentions)
Anti thbs1, by Proteintech (1 mentions)
Anti rpl10, by Proteintech (1 mentions)
Anti cd81, by Proteintech (1 mentions)
Anti cd63, by Proteintech (1 mentions)
Anti glut3, by Proteintech (1 mentions)
2 protocols using anti rbp1
1
Western Blot Analysis of Autophagy Markers
2
Exosomal Protein Expression Analysis
Check if the same lab product or an alternative is used in the 5 most similar protocols
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Western blotting was performed as described previously [9] (link). Primary antibodies against CD9, CD63, CD81, HSP90, FGFBP1, SIPA1, THBS1, TGFBI, COL6A1, RPL10, SLC2A3 (GLUT3), MYO1D, RBP1, SMOC2, GLG1, and CEMIP (Protein-Tech Group, Rosemont, IL, USA) were used.
Western blotting was performed to verify the expression of exosome marker proteins and selected exoDEPs in SW620 and SW480 exosomes. Equivalent amounts of total protein (20 μg) were resolved by 10% sodium dodecyl sulfate polyacrylamide gel electrophoresis, and the antibodies were diluted as follows: anti-CD9 (1:1000), anti-CD63 (1:1000), anti-CD81 (1:2000), anti-HSP90 (1:200), anti-FGFBP1 (1:500), anti-SIPA1 (1:2000), anti-THBS1 (1:500), anti-TGFBI (1:200), anti-COL6A1 (1:200), anti-RPL10 (1:1000), anti-GLUT3 (1:500), anti-MYO1D (1:200), anti-RBP1 (1:500), anti-SMOC2 (1:500), anti-GLG1 (1:1000), and anti-CEMIP (1:1000) (Protein-Tech Group, Rosemont, IL, USA). Proteins were detected using an enhanced chemiluminescence reagent (Santa Cruz Biotechnology, Dallas, TX, USA). Western blot signals were quantified using FluorChem E (Protein Simple, San Jose, CA, USA). All analyses were performed using western blots in at least two biological replicates.
Western blotting was performed to verify the expression of exosome marker proteins and selected exoDEPs in SW620 and SW480 exosomes. Equivalent amounts of total protein (20 μg) were resolved by 10% sodium dodecyl sulfate polyacrylamide gel electrophoresis, and the antibodies were diluted as follows: anti-CD9 (1:1000), anti-CD63 (1:1000), anti-CD81 (1:2000), anti-HSP90 (1:200), anti-FGFBP1 (1:500), anti-SIPA1 (1:2000), anti-THBS1 (1:500), anti-TGFBI (1:200), anti-COL6A1 (1:200), anti-RPL10 (1:1000), anti-GLUT3 (1:500), anti-MYO1D (1:200), anti-RBP1 (1:500), anti-SMOC2 (1:500), anti-GLG1 (1:1000), and anti-CEMIP (1:1000) (Protein-Tech Group, Rosemont, IL, USA). Proteins were detected using an enhanced chemiluminescence reagent (Santa Cruz Biotechnology, Dallas, TX, USA). Western blot signals were quantified using FluorChem E (Protein Simple, San Jose, CA, USA). All analyses were performed using western blots in at least two biological replicates.
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