Good negative control Ab to my indicator Ab (biotinylayed human IgG ) for use in a competitive ELISA?

05 October 2018 1 479 Report

I am doing a ELISA to test the competitive binding of test antibodies to human FCgR2A. My indicator antibody is biotinylated human IgG (POLYCLONAL) used at 2ug/ml. The competitive antibodies were used in the titred concentration of 10ug/ml to 0.4ug/ml. I want to know what would be a good negative control Ab to my indicator Ab (biotinylated human IgG ).

Steingrimur Stefansson

Hi Swati, like you indicated, a good negative control would be biotinylated human IgG's.

Badges
Science topic

More Swati Yadav's questions See All

Why am I getting very high background with my Secondary antibody diluted in "JUST Buffer"?

I used "JUST buffer" (20mM Sodium acetate, 20mM sodium acetate 9% sucrose, 0.01% polysorbate 20, pH 5.2) for doing my ELISA to check for FcgR2A binding of my antibodies. However, I got a very...

06 December 2018 6,749 4 View

Need a suitable ELISA protocol to test biotinylation of IgG?

I biotinylated IgG using the EZ-LINK kit (Thermo Scientific). I want to test the biotinylation using ELISA. Can someone guide me with a suitable protocol for ELISA?

09 October 2018 536 3 View

Do I need specific antibodies to stain for a fusion protein?

I am staining some brain sections stored in cryoprotectant that express a Histone H2B- GFP fusion protein that were generated ~10 years ago. I know I need to enhance signal with an anti-GFP...

07 August 2024 5,338 2 View

How to determine positive-stained cells in FACS? Use isotype or unstained control?

To compare positive and negative cell populations in flow cytometry, should I compare unstained cells with antibody stained cells? Or with the isotype control? Most papers show comparison with...

06 August 2024 6,728 6 View

Are there any commercially available Donkey anti-Alpaca secondary antibodies?

Are there any fluorescently labeled anti-Alpaca secondary antibodies raised in Donkey? So far I have only been able to find anti-Alpaca secondaries raised in Goat. Or is this not possible due to...

04 August 2024 4,255 1 View

Why my IgG antibody looks fragmented on non-reducing SDS-PAGE?

I am performing IgG purification and I have to show my results on SDS-PAGE. I use 10% tris glycine gel and prepare the samples under non reducing conditions. I am new to antibodies and therefore...

23 July 2024 6,664 6 View

How do I avoid HEK293 cell lifting during biotinylation?

I’m doing cell-surface biotinylation with HEK293 cells and after all the washing and biotin incubation steps (washing with PBS ph8, neutralizing unreacted biotin with 40mM Tris ph8) many of my...

23 July 2024 4,740 3 View

What are the strategies to Enhance IgG-Producing Plasma Cells in mice for Monoclonal Antibody Development?

I have immunized BalB/C mice with a protein using the intradermal (ID) method with Complete Freund's Adjuvant (CFA) and Incomplete Freund's Adjuvant (IFA), following a 14-day interval and three...

22 July 2024 9,160 2 View

How to label synapses in over-fixed mice brain sections (40 um) via immunohistochemistry (IHC)?

We have mice brains that were over-fixed due to old PFA used during perfusion. Thus, the synapses are no longer being labeled by the Synaptophysin (SY38) mAB. which works perfectly every other...

17 July 2024 7,767 3 View

Questions of antibodies selection?

Hi there, I'd like to use immunocytochemistry to determine a surface protein expression on mouse cell. The primary antibody I have is mouse anti mouse, and the secondary antibody I have is goat...

15 July 2024 6,008 5 View

Trouble finding an anti-Digoxigenin antibody to perform Southern Blot technique?

I'm currently using Roche anti-DIG-AKP Fab fragments to perform Southern Blot assays, but this reagent is delayed since november last year. It's anybody facing the same problem? If so, how are you...

15 July 2024 6,668 0 View

Do anyone experiment occasional staining problems with cryoprotectant containing Polyvinyl-pyrrolidone and Ethylene glycole?

Occasionally we observe lack of staining in the inner part of a skin section processed with indirect immunofluorescence. Can be due to incomplete removal of the cryoprotectan during the wash? Any...

07 July 2024 8,200 2 View