How much % lipase is required in lipase enzymatic assay?

RIA. How to measure?

When you use RIA, with a control the unknown sample with antibodies, you bare in mind the binding sites of the antibodies. However you still need to measure labelled antigen (radioactive) and not...

28 February 2021 2,133 3 View

What is the most suitable resuspension buffer for TCA/acetone precipitated protein?

I need to extract protein from fermented carbon sources for Bradford assay. Most researchers experiencing insolubility of pellet in resuspension buffer. Please assist me to select most suitable...

26 February 2021 7,129 3 View

Can I use TCA instead of TFA for protein analyis in RP-HPLC?

I am trying to analyze lipase using the reverse phase-HPLC C18 column. In the literature, 0.1 % TFA was and 60% ACN was used. Can I use TCA in place of TFA? If any other methods are available to...

25 February 2021 2,388 3 View

EMSA troubleshooting. Protein-DNA complex not migrating, cold probe lane with multiple bands. Can anyone help?

I'm running an EMSA to show the DNA binding activity of recombinant proteins versus the Wild type. Previous assays run by my research group using a different test system show the protein-DNA...

24 February 2021 8,325 1 View

What is the proper in vitro culturing method to get information about the activation of innate immune cells by VLPs?

Dear All, I am interested in studying the response of immune cells against virus-like particles (VLPs). I used the PBMC in vitro assay and treat it with the VLP using PHA as a positive ctrl while...

16 February 2021 1,337 2 View

Why does flow deformation occur at the test line and control line on nitrocellulose membranes (LFIA)?

I use 1% BSA in 10 mM phosphate buffer (pH 7.4) solution when striping my nitrocellulose membranes with antigens and antibodies. However, I always observe deformed flow when my sample or running...

16 February 2021 4,224 3 View

Can the viability assays (ex: MTT/CCK-8) measure cell proliferation? or are they solely viability assays?

I want to conduct a proliferation assay to test a chemotherapy combination on a cancer cell line. At first, I thought about the MTT assay. But now, all sources are saying that these do not measure...

14 February 2021 3,142 7 View

GST assay in microplate?

I'd like to know if anyone can provide me with a protocol for traditional GST enzymatic assay (with GSH and CDNB as substrates) in a 96 well microplate, for cell line and/or C.elegans...

11 February 2021 1,328 3 View

What lysis buffer is compatible with Bradford assay?

Hello! I am planning to use Bradford assay to measure protein content in PBMCs lysates. The problem is that Bradford assay doesn't seem to be compatible with Tris-based buffers or SDS. Are there...

10 February 2021 1,797 5 View

For PC-3 cell line. How long before irradiation do I have to synchronize my cells to G0/G1?

I am trying to do a clonogenic assay for PC-3 after irradiation. I have to synchronize my cells by serum starvation but there's a optimal time I have to do that before irradiation. been looking in...

09 February 2021 9,480 1 View

Joseph Kreit

I have understood that your objective is the lipase activity evaluation. This is a balance between the reaction time in the applied conditions and the enzyme amount. The reaction evolution as a time function must be linear. You could apply a time sufficient to measure the reaction product or the substrate disappearance. Generally, the enzyme amount is very low.