Why are my ODs the same on all my samples for ELISA?

I am running an ELISA on mouse serum that has worked beautifully in the past, and for some reason now it doesn't work for either the standard or the samples. I keep getting almost the same ODs for...

08 September 2015 6,808 6 View

Can anyone help me choose the right setting in NIS Elements software for nikon monochrome microscope camera for H&E stained tissue?

I am trying to take photos of H&E stained tissue with a monochrome Nikon microscope camera, and I can't seem to choose the right filter to apply to the image. I am using NIS Elements software....

06 July 2014 9,659 3 View

Why does my NADPH oxidase ROS curve have two peaks when stimulating with PMA?

I am stimulating mouse bone marrow PMN with PMA in a luminol ROS detection system, and reading the luminescence with a luminometer for 1 hour. I always get two peaks in my curve (an sharp,...

05 June 2013 2,541 1 View

ELISA is broken?

I have run an ELISA that normally works to detect IL-6, but today (even with all newly made working reagents) the color change reaction is very very slow. Even the standard curve is very faint. I...

04 May 2013 3,027 34 View

Buffer to disrupt platelet alpha granules for PF4 quantification?

Hello! I want to quantify by ELISA the secreted (from platelets poor plasma) and the non-secreted (from platelet lysate) PF4 before and after TRAP stimulation. I will use the ELISA from R&D...

03 March 2021 1,499 2 View

Dapi fluorescence protocol for colon cell lines ?

28 February 2021 2,534 2 View

Will a 10xHis-tag prevent anti-MBP from binding to MBP?

I have a protein with a 10xHis-MBP tag on the N-terminus. I'm fairly certain I am expressing my protein, but I'm not getting a signal on my western blot. Is there a possibility that the 10xHis tag...

25 February 2021 8,404 3 View

What's the concentration recommend for recombinant protein use in western blotting?

I'm currently working having problem with the concentration for recombinant protein use in western blot, the one been used is Recombinant Human CD19 Fc Chimera Protein-rndsystem which does not...

24 February 2021 8,514 3 View

What is the best way to manage Western blots?

Increasingly, more articles are being retracted because of duplicate or otherwise dubious Western blots. Consequently, journals are asking for original Western blots and authentication of...

23 February 2021 8,609 3 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

Can I use acetic acid with an ELITE WAX Perkin Elmer column for Gas Chromatography?

Hi! I would want to know if I can use acetic acid with an ELITE WAX Perkin Elmer column for Gas Chromatography. This column has a polyethylene glycol phase, but I can't find the manual to check...

22 February 2021 730 2 View

Diltuions for a Western blot?

I need help using the equation C1V1=C2V2 figuring out how much antibody to add to use for a western blot. The stock was origianlly 3.45 ng/ul but diluted 1:10 so now its 0.345 ng/ul. I need to run...

22 February 2021 9,901 3 View

Can anyone help me how to get the similar bands in Actin?

I run western blot on HCT-116 cell lines, I got actin band like in the attached picture. The band size is not same. Can anyone help me explain this situation and how to overcome? Thank you your...

22 February 2021 6,937 3 View

I see 2 random bands in my ERK/pERK blot? Has anyone else seen this?

I ran a western blot using 12% gel and probing for ERK/pERK (cell signaling antibodies). The green is total ERK, the Red is supposed to be phospho-ERK. I see 2 bands below the total ERK that...

22 February 2021 1,777 3 View