Bradford Assay

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This video demonstrates how to quantitate proteins using the Bio-Rad Got Protein?™ kit. Protein concentration is determined both visually by comparing samples to a set of standards and by measuring absorbance on a SmartSpec™ spectrophotometer.
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[@saweraakram3675]

What is the procedure for qualitative estimation of proteins by Bradford assay?

[@eunicetan5960]

what kind of non covalent bonding the BSA binds to protein?can you explain what happens during the incubation time,could you explain the mechanism?why does it cause a shift in maximal absorbance?

[@alisaguadarroma829]

vanderwalls and ionic bonding is taking place, the bradford reagent interacts with the protein linearly. such that more protein in the sample results in more non-covalen bonding between the dye and the protein. for more detailed info on the principle behind this mechanism see en.wikipedia.org/wiki/Bradford_protein_assay

[@eunicetan5960]

@@alisaguadarroma829 thankyou so much!!

[@sofiaolivares972]

i want to estimate the protein of pollen, does anyone have a procedure or info to help??

[@someoneOnYoutube001]

Does it matter if I use a 3.5 ml cuvette? Will it affect the absorbance?

[@felipedejesusbonillaahumad7415]

Need to read: "Lambert-Beer law" in order to understand the correlation between absorbance, concentration and dilution factor.

[@sinaerfani2375]

Is the Phosphate buffered saline a mandatory buffer to use, i don't have that buffer. If i don't use it what will happen to the pH, and is their any other buffer i can use.

[@adronung1892]

It just matters that you use the same buffer as your sample is in. So your control is just the buffer.

[@isabellek1171]

Will this work if I want to test protein concentration in soy milk?

[@isabellek1171]

@@thamizh2850 Thank you

[@isabellek1171]

@@thamizh2850 What do you mean by linear range? So the concentration of protein in the soy milk is enough for the Bradford Assay and spectrophotometer to work?

[@gowthamvenkatchalam5312]

Can we have a sprout seed with protein concentration of 55 mg/ml?

[@fatimatoheed6542]

@@thamizh2850 will this method work for the determination of protein in food such as rice or wheat?

[@fatimatoheed6542]

@@thamizh2850 thanks

[@danys9085]

thank you for having such video it was helpful

[@nwanduchikeluba4094]

For a coloured protein binding sample such as polysaccharide of inonotus obliquus, which is usually brown in colour. How should I approach it since it is not white in colour such as the milk.

[@londonberry2180]

make a 10x - 20x dilution or something, quantify it, before multiplying back by the dilution factor. Once you do the dilution, the colour should be less prominent.

[@shwethaguled9518]

I want to estimate the protein from the fish sample.. Can u suggest for me procedure plzzz...

[@TheHiddenChronicle-b9l]

yes i can suggest u procedure

[@amiraalmahrazi1816]

What is the purpose of standard,I couldn’t get it,please explain to me!?

[@londonberry2180]

basically, you can't directly quantify the amount of protein in an unknown sample. You need to do a relative quantification - which means by using a series of known protein concentration, you obtain a set of absorbance values which are arranged to form a standard curve. Then, the standard curve is used to indirectly quantify the amount of protein you have in your unknown sample based on its absorbance values.

[@hananisaad2190]

does it work for chicken?

[@prairietan]

Sponsored by Sharpie?

[@chartreuse4]

의료고 바이오과 있으면 손

[@edwinabichedid9616]

mos