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Questions tagged [assay-development]

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T cell sensitivity and persistence to specific bacterial proteins

Currently, the standard tests for Lyme Disease measure antibody production after exposure to a bacteria, Borrelia burgdorferi. Often, the tests are performed too soon after infection, before ...
Stephanie's user avatar
2 votes
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34 views

Troubleshooting direct sandwich ELISA

I am validating an ELISA measuring A1c (also called glycated hemoglobin; target for humans) for use in another species. It is a direct sandwich assay. The standard and my sample (whole blood) were ...
burphound's user avatar
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Minimum number of cycles for effective qPCR quantification?

I have been having an apparent problem with gDNA contamination in my no-reverse transcriptase (NRT) controls by the appearance of fluorescence peaks in my qPCR data. I have/am trying multiple DNase ...
jvf's user avatar
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2 votes
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Assays for protein or lipid content of insect pupae

Does anyone have any recommendations (based on personal experience) for rapid, cheap, accurate assays for estimating protein & lipid content of individual insect (mosquito) pupae? So far I've ...
arboviral's user avatar
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2 votes
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Where am I going wrong in a Reactive Oxygen Species Assay done in N9 cells?

I have followed the modified protocol that was given to me by my senior (who was using for his RAW 264.7 cells - Reference article: https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3308605/ ) for ...
srimadh's user avatar
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1 vote
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How to choose an assay for detecting virulence factors?

When looking for a protein of interest, an ELISA is being administered. I read this article about the different types of ELISAs and the advantages/disadvantages. However, I am conflicted about ...
biology1's user avatar
1 vote
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118 views

Assays to determine competitive binding versus non-competitive

I'm looking for both simple and complex assays or technologies than can be used to determine if two competing molecules are competitive or noncompetitive. I figure xray crystallography is a clear one,...
Nathan's user avatar
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1 vote
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How to calculate the LOB, LOD and LOQ of an enzyme assay

I understand how to calculate limit of blank (LOB), limit of detection (LOD), and limit of quantitation (LOQ) in the traditional way i.e., average and SD of raw analytical signal of blanks and low ...
Dave's user avatar
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Ames Assay Confusion: Aren't the odds of spontaneous revertants too low to be able to accurately test the mutagenicity of certain compounds?

I am a student conducting a test with the Ames Assay. This assay uses a strain of bacteria that has a mutation in an amino acid synthesizing operon, which doesn't allow it to synthesize its protein. ...
Kyotiq's user avatar
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What is the probe that absorbs at 450nm in the presence of NADH in this assay?

The colorimetric assays by Biovision and Sigma Aldrich seem to utilise a probe that binds to or reacts with NADH in order to cause absorbance at 450nm which can then be quantified by a ...
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