Table Basic components of the SDS-PAGE.
Sr. No. Stock Solution
1 30% Acrylamide
Bisacrylamide solution.
2 10% SDS
3 4x Resolving Gel Buffer
(1.5 M tris-HCL. pH8.8)
4 4x Stacking Buffer Solution
(0.5M Tris-HCl, pH 6.8)
5 4x Tris Glycine Running Buffer
6 10% Ammonium per sulfate (APS) 0.1g of ammonium per sulphate was
solution
Composition
and 29.0g of Acrylamide and 1g of
Methylene bisacrylamide was added in
deionized water and raised the final
volume to 100 ml with deionized water.
Added 10g SDS in distilled water and
raised the final volume to 100 ml with
distilled water.
Added 18.17 g of tris base in 50 ml of
distilled water. Adjusted the pH with
HCL drop wise and raised the final
volume up to 100 ml with distilled water
Added 12.11 g of tris base in 50 ml of
distilled water. Adjusted the pH with
HCL drop wise and raised the final
volume up to 100 ml with distilled water
Added 72.06g of glycine +15.14 g of
Tris base+5g of 1% SDS in distilled
water. Raised the final volume 0.5 Liter
with distilled water.
added in 1 ml distilled water.
8 N,N,N,NTetramethyl
Ethylenediamine (TEMED)
9 4x Sample Buffer
10 CBB R-250 Stainer
11 Destainer
Readily available for use.
4ml of glycerol + 1.2g SDS + 5ml of 4x
Stacking Buffer + 0.03g of bromophenol
blue +2ml of β-mercaptoethanol Aliquot
into 1.5ml microcentrifuge tubes. Store
at -20 °C.
Added 0.25 % of CBB in methanol:
acetic acid: water solution (50: 10:40) to
prepare the CBB stain. After that filtered
through the Whatman No. 1 filter paper.
Added methanol: acetic acid: water
solution (50ml: 10ml: 40ml) to prepare
destainer.
 Table : Dilution concentrations for BSA standard curve

Sr. Standard dilution Concentration Absorbance

No. of BSA solution (µg/ml) (595 nm)
01 0 0.00
02 50 0.02
03 100 0.04
04 200 0.15
05 400 0.18
06 800 0.27
07 1600 0.49
Bradford reagent:

Dissolved 100 mg Coomassie Brilliant Blue G-250 in 95 % ethanol (50 ml). Finally, added 100 ml of 85 % orthophosphoric acid H3PO4. When
solubilization of dye was completed, 500ml of distilled water was added to this solution. The solution was stirred gently for 3 to 4 hours for
complete mixing of dye. Filtration is done by using Whattman filter paper no.1 for removal of precipitates. Bradford reagent is very sensitive to
light, all procedure was carried out in a highly conscious way by covering reagent with aluminum foil and keeping in dark. 350ml of distilled
water was added to raise the final volume up to 1L