Using serum as sample in Western blot

More Anne Marie (Mulligan Tuttle) Gannon's questions See All

Can you use deuterated testosterone as an internal standard for ELISA?

Looking to get away from using C14 spike-in as the standard and was wondering if using deuterated testosterone is a viable option for testosterone ELISAs. Thank you!

05 June 2016 2,567 11 View

How much yield (ng/ul) do you get with RNA extraction from serum?

Just wondering what I should expect in terms of a yield in ng/ul when isolating RNA using the Qiagen miRNeasy kit from serum. Thanks!

31 December 2014 6,003 2 View

Implantation site detection method - can anyone help?

I need to stain rodent implantation sites but then want to use the uteri for histological fixation. I understand that the use of ammonium sulfide renders the tissue unusable for fixation. Any...

05 June 2013 3,107 13 View

Can anyone recommend a good protocol for C. elegans spike-in for miRNA from serum?

I'm looking for a good protocol for spike-in for rat serum miRNA quantification. I've heard that the C.elegans spike-in works well, but don't have an appropriate protocol for it. Any tips would be...

03 April 2013 9,003 10 View

Does the volume of p-npp for alkaline phosphatase make a huge difference in a sandwich ELISA?

I am running a sandwich ELISA and realised far too late that I have run low on SigmaFast p-npp tablets for my detection step. In the product information insert, Sigma suggests 200 microlitres of...

06 July 2012 1,418 6 View

Does anyone know what might be causing the smeared bands on my western blot?

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

What will be the best way to test NFkb activation via western blot?

Hi, I have lysate from human macrophages, and I wonder what will be the best way to check the NFKB activation in those cells. I was thinking p-p65 or p-ikkb./ Is that make sense?

18 July 2024 5,214 4 View

How to detect phosphorylation on Western Blots?

Having trouble detecting phosphorylated HSL. Any tips for how to prevent de-phosphorylation events or detect phosphorylation?

15 July 2024 2,055 3 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

Is there anyway to distinguish two proteins of same molecular weight?

I know mass spectrometry will work. But, is there any simple and straightforward method to distinguish between the two proteins of the same molecular weight? They are cryptochromes and...

14 July 2024 9,324 2 View

What would you do next?

Studying a specific monogenetic disease based on mutations in different individual genes. I try to recreate the disease phenotypes of donors in vitro. I knocked out a gene x with above 90%...

12 July 2024 6,313 0 View

Loading control for western blot to detect Histone H1.2?

Hello, I isolated total histones successfully from the skin primary cells by acid extraction. I wanted to detect histone H1.2 expression in different conditions by western blot. Can some one...

10 July 2024 9,165 1 View

Personal recommendations for GAPDH antibody?

Hello! I am planning a western blot experiment, and I am looking for GAPDH antibodies to order. In the past, I have experienced a lot of hit-or-miss when ordering antibodies based on just the...

07 July 2024 1,685 0 View

How can I solve this problem with my western blot?

I have this problem with my western blot for the ABCG2 antibody on nitrocellulose membrane. Help please

02 July 2024 8,715 2 View

How to troubleshoot error E1 on Biorad Powerpac Basic despite using sufficient and fresh buffer?

I have used this BIorad Powerpac Basic for 4 years now, before which it was used for another 5-6 years. Generally we addressed the E1 error by compensating with fresh buffer and/or filling the...

01 July 2024 7,104 1 View