Keloid fibroblasts were lysed with Cell Extraction Buffer (Invitrogen). The relevant cellular lysates were separated on 12% SDS‐PAGE and transferred to NC membrane, which were blocked with 5% non‐fat milk in phosphate‐buffered saline (PBS) for 2 hours and incubated with primary antibody overnight at 4°C and secondary antibody at room temperature for 1 hours. GE Healthcare ECL system was utilised to develop the signal, and the intensity of the interest bands was calculated with NIH‐Image J1.51p 22 by correcting with GAPDH expression. Primary antibodies utilised were indicated: anti‐FBXL6 (1:1000 dilution, SAB1407299, Sigma, St. Louis, MO), anti‐c‐MYC (1:1000 dilution, MA1‐980, Invitrogen), anti‐Collagen I (1:2000 dilution, PA5‐95137, Invitrogen), and anti‐GAPDH (12 000 dilution, MA1‐16757, Invitrogen).
Anti fbxl6
Manufactured by Merck Group
Sourced in United States
Anti-FBXL6 is a laboratory reagent that targets the F-box and leucine-rich repeat protein 6 (FBXL6). It is used in research applications to study the function and regulation of FBXL6, which is a component of the SCF ubiquitin ligase complex involved in protein degradation.
Automatically generated - may contain errors
Lab products found in correlation
Anti gapdh, by Cell Signaling Technology (4 mentions)
Anti flag m2, by Merck Group (4 mentions)
Anti c myc, by Cell Signaling Technology (4 mentions)
Anti skp1, by Cell Signaling Technology (4 mentions)
Anti cul1, by Santa Cruz Biotechnology (4 mentions)
Anti hsp90aa1, by Proteintech (4 mentions)
Cell extraction buffer, by Thermo Fisher Scientific (1 mentions)
Anti collagen 1, by Thermo Fisher Scientific (1 mentions)
Anti c myc, by Thermo Fisher Scientific (1 mentions)
Ecl system, by GE Healthcare (1 mentions)
5 protocols using anti fbxl6
1
Western Blot Analysis of Keloid Fibroblasts
2
Western Blot Analysis of Protein Expression
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Cells were lysed with lysis buffer (100 mM Tris-HCl, pH 6.8, 100 mM DTT, 1% SDS, 10% glycerol). Proteins were separated by 10–12% SDS-PAGE, and transferred to NC membrane. Membranes were blocked in 5% non-fat milk in phosphate-buffered saline (PBS) for 1 h before incubation with primary antibody overnight at 4 °C. Membranes were washed with and incubated with secondary antibody for 1 h. Primary antibodies used as indicated: anti-Flag M2 (1:4000 dilution, F1804, Sigma), anti-HSP90AA1 (1:1000 dilution, 13,171–1-AP, Protein tech), anti-FBXL6 (1:1000 dilution, SAB1407299, Sigma), anti-Cul1 (1:1000 dilution, sc-17,775, Santa cruz, U.S.A), anti-SKP1 (1:2000 dilution, #12248, Cell Signaling Technology, U.S.A), anti-c-Myc (1:1000 dilution, #18583, Cell Signaling Technology, U.S.A), and anti-GAPDH (1:5000 dilution, #5174, Cell Signaling Technology, U.S.A).
3
Western Blot Analysis of Protein Interactions
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Cells were lysed with lysis buffer (100 mM Tris-HCl, pH 6.8, 100 mM DTT, 1% SDS, 10 % glycerol). Proteins were separated by 10-12% SDS-PAGE, and transferred to NC membrane. Membranes were blocked in 5% non-fat milk in phosphate-buffered saline (PBS) for 1 h before incubation with primary antibody overnight at 4 °C. Membranes were washed with and incubated with secondary antibody for 1 h. Primary antibodies used as indicated: anti-Flag M2 (1:4,000 dilution, F1804, Sigma), anti-HSP90AA1
(1:1,000 dilution, 13171-1-AP, Protein tech), anti-FBXL6 (1:1,000 dilution, SAB1407299, Sigma), anti-Cul1 (1:1000 dilution, sc-17775, Santa cruz, U.S.A), anti-SKP1 (1:2,000 dilution, #12248, Cell Signaling Technology, U.S.A), anti-c-Myc (1:1,000 dilution, #18583, Cell Signaling Technology, U.S.A), and anti-GAPDH (1:5,000 dilution, #5174, Cell Signaling Technology, U.S.A).
(1:1,000 dilution, 13171-1-AP, Protein tech), anti-FBXL6 (1:1,000 dilution, SAB1407299, Sigma), anti-Cul1 (1:1000 dilution, sc-17775, Santa cruz, U.S.A), anti-SKP1 (1:2,000 dilution, #12248, Cell Signaling Technology, U.S.A), anti-c-Myc (1:1,000 dilution, #18583, Cell Signaling Technology, U.S.A), and anti-GAPDH (1:5,000 dilution, #5174, Cell Signaling Technology, U.S.A).
4
Western Blot Analysis of Protein Interactions
Check if the same lab product or an alternative is used in the 5 most similar protocols
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Cells were lysed with lysis buffer (100 mM Tris-HCl, pH 6.8, 100 mM DTT, 1% SDS, 10 % glycerol). Proteins were separated by 10-12% SDS-PAGE, and transferred to NC membrane. Membranes were blocked in 5% non-fat milk in phosphate-buffered saline (PBS) for 1 h before incubation with primary antibody overnight at 4 °C. Membranes were washed with and incubated with secondary antibody for 1 h. Primary antibodies used as indicated: anti-Flag M2 (1:4,000 dilution, F1804, Sigma), anti-HSP90AA1
(1:1,000 dilution, 13171-1-AP, Protein tech), anti-FBXL6 (1:1,000 dilution, SAB1407299, Sigma), anti-Cul1 (1:1000 dilution, sc-17775, Santa cruz, U.S.A), anti-SKP1 (1:2,000 dilution, #12248, Cell Signaling Technology, U.S.A), anti-c-Myc (1:1,000 dilution, #18583, Cell Signaling Technology, U.S.A), and anti-GAPDH (1:5,000 dilution, #5174, Cell Signaling Technology, U.S.A).
(1:1,000 dilution, 13171-1-AP, Protein tech), anti-FBXL6 (1:1,000 dilution, SAB1407299, Sigma), anti-Cul1 (1:1000 dilution, sc-17775, Santa cruz, U.S.A), anti-SKP1 (1:2,000 dilution, #12248, Cell Signaling Technology, U.S.A), anti-c-Myc (1:1,000 dilution, #18583, Cell Signaling Technology, U.S.A), and anti-GAPDH (1:5,000 dilution, #5174, Cell Signaling Technology, U.S.A).
5
Western Blot Analysis of Protein Targets
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Cells were lysed with lysis buffer (100 mM Tris-HCl, pH 6.8, 100 mM DTT, 1% SDS, 10 % glycerol).
Proteins were separated by 10-12% SDS-PAGE, and transferred to NC membrane. Membranes were blocked in 5% non-fat milk in phosphate-buffered saline (PBS) for 1 h before incubation with primary antibody overnight at 4 °C. Membranes were washed with and incubated with secondary antibody for 1 h. Primary antibodies used as indicated: anti-Flag M2 (1:4,000 dilution, F1804, Sigma), anti-HSP90AA1 (1:1,000 dilution, 13171-1-AP, Protein tech), anti-FBXL6 (1:1,000 dilution, SAB1407299, Sigma), anti-Cul1
(1:1000 dilution, sc-17775, Santa cruz, U.S.A), anti-SKP1 (1:2,000 dilution, #12248, Cell Signaling Technology, U.S.A), anti-c-Myc (1:1,000 dilution, #18583, Cell Signaling Technology, U.S.A), and anti-GAPDH (1:5,000 dilution, #5174, Cell Signaling Technology, U.S.A).
Proteins were separated by 10-12% SDS-PAGE, and transferred to NC membrane. Membranes were blocked in 5% non-fat milk in phosphate-buffered saline (PBS) for 1 h before incubation with primary antibody overnight at 4 °C. Membranes were washed with and incubated with secondary antibody for 1 h. Primary antibodies used as indicated: anti-Flag M2 (1:4,000 dilution, F1804, Sigma), anti-HSP90AA1 (1:1,000 dilution, 13171-1-AP, Protein tech), anti-FBXL6 (1:1,000 dilution, SAB1407299, Sigma), anti-Cul1
(1:1000 dilution, sc-17775, Santa cruz, U.S.A), anti-SKP1 (1:2,000 dilution, #12248, Cell Signaling Technology, U.S.A), anti-c-Myc (1:1,000 dilution, #18583, Cell Signaling Technology, U.S.A), and anti-GAPDH (1:5,000 dilution, #5174, Cell Signaling Technology, U.S.A).
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