Protein lysates from fibroblasts were prepared using RIPA buffer, separated on Criterion Precast Gels (Bio-Rad) 4–15% Tris-HCl, followed by transfer to PVDF Membrane (Bio-Rad). Membranes were probed with the following primary antibodies in TBS-T and 5% milk for 12 to 16 hours: Anti-Notch1 (R&D AF33471), Anti-Notch3 (V1662) 1ug/ml, GAPDH (1:1000, Cell Signaling). Secondary antibodies (1:10,000) were from Jackson ImmunoResearch. Chemiluminescent detection reagents were from Bio-Rad.
Chemiluminescent detection reagents
Manufactured by Bio-Rad
Chemiluminescent detection reagents are a class of laboratory equipment used to detect and quantify specific proteins or molecules in biological samples. They work by generating a luminescent signal when a target analyte is present, which can be measured using a specialized detection instrument.
Automatically generated - may contain errors
3 protocols using chemiluminescent detection reagents
1
Quantitative Notch Signaling Analysis
2
Quantitative Notch Signaling Analysis
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Protein lysates from fibroblasts were prepared using RIPA buffer, separated on Criterion Precast Gels (Bio-Rad) 4–15% Tris-HCl, followed by transfer to PVDF Membrane (Bio-Rad). Membranes were probed with the following primary antibodies in TBS-T and 5% milk for 12 to 16 hours: Anti-Notch1 (R&D AF33471), Anti-Notch3 (V1662) 1ug/ml, GAPDH (1:1000, Cell Signaling). Secondary antibodies (1:10,000) were from Jackson ImmunoResearch. Chemiluminescent detection reagents were from Bio-Rad.
3
Chondrocyte Protein Expression Analysis
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Protein lysates of articular chondrocytes were prepared in RIPA buffer, separated on Criterion precast gels (Bio-Rad) 4-15% Tris-HCl, and then transferred to PVDF membranes (Bio-Rad). Membranes were probed in TBS-T and 5% milk for 12-16 hours with primary antibodies: EGFR (1:2000 A nity), β-actin (1:50000 abclonal). Secondary antibody (1:5000) was from a nity. Chemiluminescent detection reagents were from Bio-Rad.
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