Total protein was extracted from tissue or cell by radioimmunoprecipitation assay (RIPA) lysate containing PMSF (P0013C, Beyotime, Shanghai, China) followed by protein concentration detection. The samples were then transferred to a polyvinylidene fluoride membrane after sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE). After that, the membrane was blocked with 5% skim milk powder at room temperature for 1 h and incubated overnight at 4 °C with diluted primary rabbit antibodies against ELK4 (1:1000, ab86002, Abcam, Cambridge, MA, USA), KDM5A (1:5000, ab194286, Abcam), PJA2 (1:800, ab131118, Abcam), KSR1 (1:1200, ab68483, Abcam), and GAPDH (ab9485, Abcam; internal reference). The membrane was then incubated with HRP-labeled goat anti-rabbit against IgG H&L (ab97051, 1:2000, Abcam) for 1 h. Subsequently, the membrane was developed by the enhanced chemiluminescence kit (BB-3501, Ameshame, UK). The images were taken by Bio-Rad image analysis system (Bio-Rad Laboratories, Hercules, CA, USA) and analyzed by Quantity One v4.6.2 software.
Ab68483
Manufactured by Abcam
Sourced in United States
Ab68483 is an antibody reagent. It is intended for use in various life science research applications.
Automatically generated - may contain errors
Lab products found in correlation
Ab194286, by Abcam (1 mentions)
Ab97051, by Abcam (1 mentions)
Ab86002, by Abcam (1 mentions)
Ab9485, by Abcam (1 mentions)
Image analysis system, by Bio-Rad (1 mentions)
Ab32376, by Abcam (1 mentions)
Anti actin antibody a2066, by Merck Group (1 mentions)
Ab221611, by Abcam (1 mentions)
Horseradish peroxidase conjugated goat anti rabbit igg, by Jackson ImmunoResearch (1 mentions)
2 protocols using ab68483
1
Protein Expression Analysis by Western Blot
2
Comprehensive Antibody Validation for Ras-MAPK Signaling
Check if the same lab product or an alternative is used in the 5 most similar protocols
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Anti-KSR1 (ab68483) and anti-PKCα (ab32376 used for immunoblot analysis; ab221611 used for immunoprecipitation) antibodies were from Abcam; anti-H-Ras (A19619), anti–pan-Ras (A4735), anti-RasGRP1 (A10495), and anti-PKCε (A2110) antibodies were from Abclonal; anti-KSR2 (H00283455-M08) antibody was from Abnova; anti-A-Raf (75804S), anti-B-Raf (14814S), anti–phospho-Ser259 C-Raf (9421), anti–phospho-Ser338 C-Raf (9427), anti-cyclin D1 (2978), anti-Gapdh (5174), anti–pan-Ras (3339S), anti-pERK (9106 and 4370), anti-pMEK (9121), anti–total ERK (9102), anti–total MEK (9122), anti-SOS1 (D3T7T), and anti-pRSK(S380) (9341) antibodies were from Cell Signaling Technology; anti-C-Raf (610151) and anti-p21Cip1 (556430) antibodies were from BD Pharma; anti-Id1 (BCH-1/195-14-50) antibody was from BioCheck; anti-pRSK(S380) (04-418) was from Millipore; anti-p21Cip1 (NBP2-29463) antibody was from Novus Biologicals; anti-RasGRP3 (sc-271068), anti-SOS2 (sc-393667), and anti-PKCδ (sc-8402) antibodies were from Santa Cruz Biotechnology; and anti-actin (A2066) antibody was from Sigma–Aldrich. Secondary antibodies were horseradish peroxidase–conjugated goat anti-rabbit IgG (AP132P, Millipore), horseradish peroxidase–conjugated goat anti-rabbit IgG (111-035-144, Jackson Immunoresearch Laboratories), and horseradish peroxidase–conjugated goat antimouse IgG (170-6516, Bio-Rad).
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