BRADFORD ASSAY

Calculation

Formulations:

Absorbance of BSA = A595 nm

Raw Data = Average of A595 nm of 3 wells ÷ 3

Case: A1+A2+A3 ÷ 3

In this case, 0. 365+0. 374+0. 453 ÷ 3 sama dengan. 397

Remedied Data =( (A595 nm well) – BackGround) or perhaps (Raw Info – Background) Background = negative control = Liverpool Reagent + No Proteins Background of the standard curve = A595 nm of well ‘A' =. 397 We employed well ‘A' as each of our negative control.

INFO

RAW INFO CORRECTED INFO BSA (µg)| A595 nm

0|. 397

2 . 5|. 795

some. 0|. 743

5. 0|. 863

6. 0|. 742

7. 5|. 958

twelve. 0|. 995

BSA (µg)| A595 nm

0| zero

2 . 5|. 398

4. 0|. 346

5. 0|. 466

6th. 0|. 345

7. 5|. 561

12. 0|. 553

Standard Competition Raw Data

(Figure 1)

Normal Curve-Corrected Info

(Figure 2)

The numbers, 1 and 2, demonstrate relationship of Bovine Serum Albumin concentration and Ingestion of light at 595 nm. Figure you shows the typical curve of Raw Data and number 2 reveals the standard curve- Corrected Info. Bovine Serum Albumin or perhaps BSA was weighed and dissolved in water to obtain a certain share concentration. Normal water was used as being a diluent. Quantitative dilutions of the stock were performed to own indicated concentration, from 0. 0 to 10. 0 μg/μl, of BSA in a final volume of 25. 0 μl. The concentration of BSA is definitely shown in ‘X-axis'. The absorption of light by every concentration for a wavelength of 595 nm was determined which has a spectrophotometer. The absorbance Ideals of each focus is shown in ‘Y-axis'. From the both figures, we could notice that because the attention level of BSA increases absorbance value at wavelength of 595 nm also raises. In numbers 1 and 2, you observe a decrease in absorbance value at BSA concentrations, four. 0 µg and 6. 0 µg. This could be as a result of bad pipetting technique.

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