Is CD44 degraded in lysosome together with hyaluronan after CD44 mediated endocytosis of hyaluronic acid?

Also when RHAMM binds hyaluronic acid, they get internalized, will RHAMM also be degraded? Or both CD44 and RHAMM will be transferred back to the cell membrane? Asking for breast cancer cell line...

01 March 2021 8,169 2 View

How to manage western blot for proteomics data validation?

Hi Everyone, I am a bigginer in western blotting and I am trying to use it for validation of my proteomics data. I have: two regulated proteins with molecular mass of 40 and 73 kDa one control...

23 February 2021 6,671 4 View

I can detect my protein but the encoding DNA sequence has stop codons within. Should I ignore the stop codons?

I constructed a synthetic DNA library (scFv, VH-VL orientation) with a 3' reporter and 8x histag. I cloned and expressed this gene following which I performed an ELISA. The ELISA results suggested...

19 February 2021 7,006 5 View

(western blot) can methanol damage my antibody signal?

I accidentally rinsed with methanol after adding my primary antibody to my membrane. I noticed after ~10 seconds and immediately replaced it with PBST (3x15 min washes). Will my antibody signal be...

12 February 2021 5,824 3 View

What is the reason, we got both test and control band on LFA in negative sample?

I am working on Lateral flow assay for virus detection by carbon conjugated polyclonal antibody. We got a high intense band in both test and control line on LFA in negative sample used as buffer....

11 February 2021 6,558 2 View

Background and spots in Western Blot membrane?

Do you know why my membrane looks like this? I did this western for Anti-hnRNP D/AUF1 antibody ab50692, as primary antibody in 1:1000 overnight at 4c and Anti-rabbit IgG HRP linked antibody...

07 February 2021 5,591 4 View

During an anti-His capture ELISA has anyone had issues with histidine residues from secondary antibodies non-specifically binding?

Hi, Hope you can help. Currently been running some sandwich ELISA assays where I coat with anti-His, block and then capture His-tag protein and then sera and then anti-bovine monoclonal...

07 February 2021 7,101 1 View

How do I keep my prefixed (PFA) frozen tissue sections from lifting off the slide?

I'm trying to stain mouse lung tissue IHC-IF I fix in 4% PFA overnight immediately upon harvesting tissues Then dehydrate/cryoprotect in sucrose gradients 15% to 30% Then freeze in OCT media...

06 February 2021 2,336 3 View

Antibody dilution for IHQ in OCT frozen tissue is more similar to IHC in Paraffin-embedded samples or to Immunocytochemistry?

31 January 2021 8,678 3 View

MHC Rabbit Genome Annotation? Domain annotation of each MHC Rabbit Gene?

I want to find a database(s) or a method where I can find and bulk download all the rabbit MHC genes and domain annotation. In humans using Ensembl, all MHC genes have names and domains but in...

28 January 2021 7,722 2 View