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Instructions for use of Coomassie brilliant blue assay kit for protein content determination

 Last Update: 2022-08-30
 Source: Internet
 Author: User

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Coomassie brilliant blue method for protein content determination kit instructions
spectrophotometry 50 tubes/48 samples

Note: Before the formal determination, select 2-3 samples with large expected differences for pre-determination
.

Determination meaning: Determination principle: Self-provided instruments and supplies: Reagent composition and preparation: Extraction of soluble protein in the sample:

Liquid samples: Clear liquid samples can be measured directly
.
Tissue sample: according to the ratio of tissue mass (g): extract volume (mL) to 1:20 (it is recommended to weigh about 0.
05 g of tissue, add 1 mL of extract (self-prepared, choose enzyme extraction buffer or distilled water or physiological brine))

Bacteria and fungi: according to the number of cells (10 ⁴ ): the ratio of the volume (mL) of the extract solution is 500~1000:1 (it is recommended to add 1mL of extract solution to 5 million cells), and ultrasonically break the cells in an ice bath (power 300w, ultrasonic for 3 seconds) , the interval is 7 seconds, the total time is 3 minutes); then, centrifuge at 8000g, 4 ℃ for 10 minutes, take the supernatant and put it on ice for testing
.

Measurement operation:

Reagent (μL)	Assay tube	blank tube
sample	500
distilled water		500
Reagent one	500	500
After mixing, measure the absorbance at the wavelength of 620 nm, △A=A measure-A blank .

Note: The blank tube only needs to be measured once
.
Calculation formula:
²

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