I could see bands while staining with Ponceau.
The antibody solutions and the incubation times are as per requirement.
Last time I added 2ml per well in a 6 well plate with 400,000 cells per well. This is the normal standard that i have seen in manuals but it gave me a low yield of cells on the day of...
07 August 2018 8,796 0 View
I did my gel running and transfer like i always do but this time when i stained the nitocellulose membrane with ponceau , the protein sample lanes apeared different towards the end . They appeared...
05 June 2018 8,565 7 View
Hello all, I have been doing Western blotting now for sometime. I make cell lysates from my transfected cells which express a certain flavoprotein. My electrophoresis and gel transfer happens...
31 December 2017 3,130 0 View
It is an AAV vector
08 September 2017 2,703 4 View
I got these smeared bands quite often lately. We typically run the gel at 140V with a 10-12% gel and do a wet transfer at 220 mA for 1.5 hr in cold room. We also noticed some dirty spots/dots (see...
10 August 2024 7,480 3 View
Can anyone share any reference that gives mouse specific CD3 antibody sequence. I have been searching for this since few days and no luck. Thank you very much
08 August 2024 9,157 4 View
I'm working on selecting antibodies against a recombinant protein that has a His-tag. My idea is to first bind the recombinant protein to a HisTRAP column and then use this column for an affinity...
07 August 2024 505 3 View
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
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
Hi all, I was just wondering if anyone has experience with multiplexing a mouse monoclonal primary and a rat primary. I'm trying to multiplex by incubating them in the same well but was told by a...
06 August 2024 9,710 1 View
Hi guys If anyone is currently working on aging cells, you guys would like to give me some advice. I'm testing against biomarker (SA-beta-Gal), I encountered a false positive in the control group...
02 August 2024 6,735 1 View
We have a lateral flow test on Sartorius 140 NC. The conjugate is gold-monoclonal antibody. Using different control lines (GAM, protein A, protein G) we get a very strong leading edge with reduced...
31 July 2024 4,510 3 View
Hello everyone, I am currently using washed human platelets to stain Annexin V as a procoagulant marker. Additionally, I am staining with PerCP-CD61 to identify platelet cells. So far, I have...
29 July 2024 8,624 1 View
I am running a western on HIV envelope protein. The size is around 50Kda and I am using rat anti-HA for the first stain and a second stain of mouse anti-CA. Primary antibody dilution is 1: 1000...
29 July 2024 6,484 5 View