Is RIPA buffer compatible with BCA assay?

Is RIPA buffer compatible with BCA assay?

RIPA Buffer is compatible with the Thermo Scientific™ Pierce™ BCA Protein Assay Kit (Cat. No. 23225). Note: If desired, add protease and phosphatase inhibitors to the RIPA Buffer immediately before use.

Can you vortex BSA?

1 Answer. Never vortex the BSA solution. The best way to prepare a BSA solution is to put the BSA powder on the buffer solution (or water) instead of the other way round. Tap or rock the tube to dissolve the BSA.

How long is BSA good for?

Lyophilized BSA although stable at room temperature for 3 weeks, should be stored 2-8°C. Upon reconstitution BSA should be stored at 4°C between 2-7 days and for future use below -18°C.

Does DTT interfere with BCA assay?

Some substances, including most reducing agents, interfere at even low concentrations in the BCA Assay. When disulfide reducing agents such as dithiothreitol (DTT) and 2-mercaptoethanol (2-ME) are the sole cause of the incompatibility, reagents can be used to chemically block the interfering sulfhydryl groups.

Does glycerol interfere with BCA assay?

The following substances interfere even at very low concentrations: ascorbic acid, catecholamines, creatinine, impure glycerol, hydrogen peroxide, hydrazides, iron, certain lipids, melibiose, phenol red, impure sucrose, tryptophan, tyrosine, uric acid.

Is RIPA buffer compatible with Bradford assay?

The RIPA buffer contains a mix of linear (SDS) and bulky (Igepal/Triton, deoxycholate) detergents, which makes it difficult to use in the Bradford assay.

Where is BSA stored?

In its lyophilized form, and when prepared in an area with low contamination, the BSA can be stored at room temperature. However, once in solution, the recommended storage temperature is 2-8 degrees C to best prevent microbiological activity.

How do you get 0.1 BSA on PBS?

To prepare 0.1% (w/v) BSA fraction V in PBS, resuspend 10 mg of BSA fraction V (Sigma) in 10 ml of Ca2+/Mg2+-free PBS to a final concentration of 0.1% (w/v).

What is a BSA standard curve?

A standard curve is a plot of absorbance vs. a varying amount of some known concentration of protein. Two common proteins used for standard curves are bovine serum albumin (BSA) and an immunoglobin (IgG).

How do you calculate Bradford assay concentration?

Perform the assay and calculate the standard (see below). The result should be around 0.5mg/ml. To calculate the concentration of the undiluted, unknown sample, simply multiply by the dilution factor. So, 0.5 x 10= 5mg/ml.

What does BCA assay measure?

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.

Can BSA be filter sterilized?

Dissolve 10 g BSA (e.g., Sigma) in 100 ml H2O. Filter sterilize using a low‐protein‐binding 0.22‐µm filter. Store indefinitely at 4°C.

What is PBS network?

The Public Broadcasting Service (PBS) is an American public broadcaster and television program distributor.

How do you get 2% BSA on PBS?

2% dry skimmed milk in PBS For one plate dissolve 0.2 g of powder in 10 mL of 1X diluted PBS. 5. 1% BSA in PBS For one plate dissolve 0.2 g of BSA in 20 mL of 1X diluted PBS. 6.