TECHNOLOGY

1. Why Is BCA Total Protein Quantification Required for ELISA Detection of Tissue Homogenate Supernatant and Cell Lysates?​​

• ① Reduce Errors: Variability in sample volume and protein extraction efficiency during tissue homogenization/cell lysis can lead to inconsistencies. BCA quantification ensures uniform total protein concentrations across samples, improving ELISA accuracy.
• ② Enhance Reliability: Standardizing total protein levels across samples ensures ELISA results are comparable and reproducible.
• ③ Journal Requirements: High-impact journals often mandate protein quantification data to validate experimental rigor and reproducibility. Thus, BCA quantification is strongly recommended prior to ELISA.

2. Do Tissue Homogenate Supernatant and Cell Lysates Need to Be Diluted to the Same Concentration (μg/μL) Before ELISA Loading?​​

• When testing the supernatant of tissue homogenates and cell extracts, there is no need to dilute them to the same concentration (ug/ul) before loading the samples. Just handle the tissue homogenate and cell extract in the recommended way and conduct the BCA total protein quantification experiment. The target concentration measured by ELISA should be corrected for the ratio of the total protein concentration, and the final result should be expressed in the mass unit /mg prot (such as pg/mg prot or ng/mg prot, etc.) indicated on the ELISA kit standard.