Comparing data from MTT Assay and IC5O?

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

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

Can any suggest me good method of GC MS for the separation of Flavonoids?

I prepared the ethyl acetate, ethanol and butanol fraction from the ethanol extract of seed for isolation of Flavonoids. Amongst them ethanol fraction showing shinoda test positive for polar...

13 February 2021 8,263 3 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

Shin Murakami

The following RG discussion might be helpful:

https://www.researchgate.net/post/How-can-I-calculate-IC50-from-mtt-results

Mavis Lim

The lower the IC50, the higher the toxicity level is. Because at the lower concentration of the extract already able to exert 50% of killing. Data got from MTT Assay, can use SPSS to do IC50 prediction.

Mani Sharma

The lowered IC 50 values of formulation system with organic solution might be due to the toxicity caused or enhance by ethanol as compared to buffer or aqueous medium. Thus, showing lower IC 50 values(50% cell kill conc.)in organic based formulations as compared to that of aqueous. Kindly refer the attached research paper for detail description on same.

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