What Is BCA Reagent?

BCA is

a weak acid composed of two carboxylated quinoline rings

and serves the purpose of the Folin reagent in the Lowry assay, namely, to react with the product (Cu

^{1 +}

ions) of complexes between copper ions and peptide bonds to produce a purple end product that strongly absorbs at 562 nm (Smith et al., 1985).

What does BCA stand for in chemistry?

BCA stands for

before-change-after

. Students start by filling in the quantity of reactants that are present before the chemical reaction happens.

How does BCA reagent work?

The BCA protein assay is used

for quantitation of total protein in a sample

. The principle of this method is that proteins can reduce Cu

^{+ 2}

to Cu

^{+ 1}

in an alkaline solution (the biuret reaction) and result in a purple color formation by bicinchoninic acid.

What is the principle of BCA?

The principle of the bicinchoninic acid (BCA) protein assay1

relies on the formation of a Cu2+-protein complex under alkaline conditions, followed by reduction of the Cu2+ to Cu1+

. The amount of reduction is proportional to the amount of protein present.

Is BCA reagent light sensitive?

Protect from light: It is recommended to keep reagents within the kit box to minimize exposure to light. Directions for Use: Please see attached protocol. Background: Bicinchoninic Acid (BCA) is capable of forming an intense purple complex with cuprous ion, Cu1+, in an alkaline environment.

Does blood interfere with BCA assay?

All Answers (3) Hi Chris, hemoglobin absorbs light around the same wavelength used for the BCA assay. So yes,

presence of RBCs (hemoglobin) could interfere with the accuracy of the assay

.

Why is BSA used in BCA assay?

BSA is used because of

its stability to increase signal in assays

, its lack of effect in many biochemical reactions, and its low cost, since large quantities of it can be readily purified from bovine blood, a byproduct of the cattle industry.

How do you write a BCA table?

Step 1: Write the balanced equation. Step 2: Make sure you have moles for your starting value. (Convert from grams to moles using the molar mass if needed.) Step 3: Insert the starting moles into the BCA Table and complete the “B” Row.

Why is stoichiometry difficult?

Stoichiometry can be difficult

because it builds upon a number of individual skills

. To be successful you must master the skills and learn how to plan your problem solving strategy. Master each of these skills before moving on: Calculating Molar Mass.

How can I do BCA method?

1. Prepare samples containing 0.2 to 50 micrograms protein in microliters.
2. Add 1 ml SWR to each 20 microliters sample and mix. Incubate 30 min. at 60 degrees C.
3. Cool the samples and read at 562 nm. Color will be stable for at least one hour.

What does BCA assay react with?

The BCA Protein Assay combines

the protein-induced biuret reaction

(see above) with the highly sensitive and selective colorimetric detection of the resulting cuprous cation (Cu1+) by bicinchoninic acid (BCA). … First is the biuret reaction, whose faint blue color results from the reduction of cupric ion to cuprous ion.

How does a BCA assay detect proteins?

Compared to other methods, the BCA assay is one of the most sensitive (it can detect proteins at concentrations as low as 5 ug/mL). … The protein concentration is

determined by the amount of dye in the blue ionic form measured by the absorbance of the solution at 595 nm using a spectrophotometer

[24].

Why is BSA used in biuret test?

An aqueous solution of

bovine serum albumin

(

BSA

) is commonly

used

as a standard in

testing

because of its stability in

testing

, low cost, and is readily available as a byproduct from bovine blood.

Is BCA better than Bradford assay?

Historically,

the BCA method is more sensitive than the Bradford method

, because the first method is based on protein-copper chelation and secondary detection of the reduced copper. Whereas the Bradford method is based on protein-dye binding and colour shift from 465 to 595 nm.

How accurate is Bradford assay?

The Bradford assay is very fast and uses about the same amount of protein as the Lowry assay. It is

fairly accurate

and samples that are out of range can be retested within minutes. … It is sensitive to about 5 to 200 micrograms protein, depending on the dye quality.

Is RIPA buffer compatible with BCA assay?

RIPA Buffer is compatible with

the Thermo ScientificTM PierceTM BCA Protein Assay

Kit (Cat. No. 23225). Note: If desired, add protease and phosphatase inhibitors to the RIPA Buffer immediately before use.