In my Fluorospot Assay to detect antigen secreting cells I coated few well with only PBS without antigen. Why I am getting Spots in those wells?

More Someshwar Jha's questions See All

What could be reason for getting such type of graph for complex impedance in case of polymer blend films?

Its effect appears in real part of ac conductivity also. Can anyone suggest any reason behind it?

26 February 2021 3,487 3 View

Does behavior (shy vs bold; rare vs abundant) and flock size (small vs large) of bird species play role in niche partitioning among sympatric species?

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Which type of polymer is used in base boards of Printed Circuit Boards (PCBs)?

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25 May 2020 9,640 15 View

Dear Peers! Could you please oblige me with your knowledge on vulture?

I am looking for proverbs and idioms or any short story mentioning vulture in any language or locality with English translation.

24 February 2020 2,004 9 View

I am supposed to develop a psychometric employability assessment. Any literature or advice that could help me? Also, how do I ensure honest answers?

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Which funding agency in India will grant traveling allowance for international conference in finance ??

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Can anyone suggest me a solver for binary linear multiplicative programming problem?

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How can we estimate Nitrogen (N) in sugarcane field using satellite imagery?

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How can we download the all the clean drug like compounds from Zinc12 database?

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Bacterial Growth Concern ?

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RIA. How to measure?

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How T cells recognise non-peptide antigens?

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Will it be possible to get IC:50 if we target cancer cell line with antibody drug conjugate (ADC) that have 50% less antigen positive cells ?

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Can I heat my ELISA at 37C?

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How to characterize epitopes within a peptide sequence?

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How can I do antigen retrieval for proliferative markers (MCM2, Ki67 etc) While preserving a sensitive endogenous reporter?

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Can anyone walk me through IHC/IF Staining of Cryosectioned Brain Tissues?

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No title

It is important for me to know the opinion from your point of view as researchers.

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Any recommendation on SaCas9 antibody?

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Can anyone share a protocol please for detecting presence of a hypothetical antibody against a known antigen in mice plasma?

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Wolfgang Schechinger

Does "complete RPMI" mean with or without serum?

Someshwar Jha

With serum

Ivan Pavlovic

Sorry, do you mean antibody secreting cells? Is this an IgG, IgA or IgM assay? I have had the same issues with the IgG/IgM FluoroSpot assay and it seems that this issue is attributed to the fact that immunoglobulins, especially IgM, are quite sticky. What you might want to try is to increase the amount of times you wash the wells after the cells have been removed. This is going to be my next attempt to minimize the background.