A modified coomassie brilliant blue G 250 staining method for the detection of chitinase activity and molecular weight after polyacrylamide gel electrophoresis

SUMMARY Isoelectric focusing in polyacrylamide gel followed by staining with Coomassie brilliant blue was used for the densitometric estimation of ovine prolactin standard and canine pituitary prolactin. The results were compared with those obtained by conventional polyacrylamide gel electrophoretic procedures and isoelectric focusing was found to be a valid technique for the estimation of prolactin and to offer greater technical convenience. The mobility of ovine and canine prolactin was similar in isoelectric focusing and gel electrophoresis. The isoelectric point of ovine and canine prolactin was found to be 6·17 and 6·61 respectively. Constant levels of prolactin were found in the pituitaries of bitches at oestrus.

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Effect of pre-stain methanol washing on the sensitivity of CBBR-250 staining in SDS-polyacrylamide gels

Sample Preparation ◽

10.1515/sampre-2015-0001 ◽

2015 ◽

Vol 2 (1) ◽

Author(s):

Rashid Saleem ◽

Absar-ul Hasnain ◽

Riaz Ahmad

Keyword(s):

Molecular Weight ◽

Acetic Acid ◽

Staining Method ◽

Muscle Protein ◽

Brilliant Blue ◽

Low Molecular Weight ◽

Polyacrylamide Gels ◽

Coomassie Brilliant Blue ◽

Protein Research ◽

Coomassie Brilliant

AbstractCoomassie Brilliant Blue R-250 (CBBR-250) is the most frequently used dye stain in protein research. However, relatively poor sensitivity and long application durations are the major limitations of this stain. In the present study, we have investigated the effect of pre-stain methanol washing on the sensitivity of the conventional CBB staining method. Concentrations of methanol ranging from 5 to 30% were prepared in 5% acetic acid. Pre-stain washing of SDS-gels using acetic acid (5%) and methanol were assessed at various timings and temperatures. Our results demonstrate that pre-stain washing by methanol greatly improves the sensitivity of the CBB stain and facilitates the visualization of low molecular weight components present in a complex muscle protein, actomyosin. The outcome of this study also revealed duration of pre-stain washing and temperature as other factors that determine the sensitivity of CBB staining method. Therefore, it is suggested that pre-stain washing with methanol may improve the sensitivity of CBB stain in SDS-PA gels.

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Improved Measurement of Monoclonal Paraproteins in Serum Using Agarose Gel Electrophoresis

Annals of Clinical Biochemistry International Journal of Laboratory Medicine ◽

10.1177/000456328702400111 ◽

1987 ◽

Vol 24 (1) ◽

pp. 73-76 ◽

Cited By ~ 6

Author(s):

P M Dennis ◽

B Biegler ◽

R Papas

Keyword(s):

Cellulose Acetate ◽

Gel Electrophoresis ◽

Brilliant Blue ◽

Agarose Gel ◽

Agarose Gel Electrophoresis ◽

Brilliant Blue G ◽

Cellulose Acetate Electrophoresis ◽

Coomassie Brilliant Blue ◽

Ponceau S ◽

Coomassie Brilliant

Serum paraproteins can be accurately and precisely measured by means of agarose gel electrophoresis and densitometry at 520 nm after staining with Coomassie Brilliant Blue G. The results obtained for this method agree closely with the quantity of paraprotein recoverable from preparative agarose gel electrophoresis. The method is preferable to cellulose acetate electrophoresis stained with Ponceau S.

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