Can anyone recommend a protocol for isolation of neutrophils from mouse liver?

Can anyone recommend a protocol to isolation neutrophils from mouse liver? Thanks Ilyas

06 July 2015 3,451 3 View

Does anyone have Serum ethanol assay?

07 August 2014 8,282 6 View

How to prepare secondary antibody from lyophilized antibody?

So I'm new to lab research and learning Western Blots these days. I ordered my secondary antibody and it arrived in a powdered form. Here is the...

27 January 2021 5,402 3 View

Western blot: what causes patchy, uneven background?

Sometimes my western blots have very uniform/even background, other times they're very uneven and patchy like in the attached image. Why is this? I thought my blots were drying out during...

05 December 2020 8,656 5 View

Licor Intercept Blocking Buffer -- sterility and reusing antibody dilutions?

In the past, I've always used Licor's old blocking buffer that included sodium azide. I would dilute my antibodies in the buffer, stain, re-collect the dilution, and re-use it a few times. The...

28 November 2020 5,795 3 View

Western blot: adjusting volume of buffer for blocking/staining?

For western blots, I normally stain with 10 mL buffer with 1:1,000 dilution of my specific antibody. Recently, there is an equipment change in my lab and 10 mL isn't enough volume to cover the...

27 October 2020 2,749 7 View

Are there Hif1a antibodies that work reliably for immunofluorescence on frozen sections?

A few years back I tried several available Hif1a antibodies on frozen sections of heart and lung. None of them worked very well. Most worked well for immunoblots. Have there been any advances in...

12 October 2020 7,727 2 View

What is wrong with this western blot?

Dear fellow researchers: I've recently started performing WB, mostly with molecular weight markers to test the equipment and optimize I/V conditions. This time I also ran protein lysates from two...

12 August 2020 3,943 11 View

On my western blotting/ECL, using anti-alpha tubulin, aside the 55kDa alpha tubulin band, extra band appears at 35 kDa, wondering wht this band is?

When using abcam anti-alpha-tubilin on immunoblotting developed on ECL to probe tubulin on mice splenocytes, I see a clear band at 55kDa but there appears consistently another band at about 35kDa,...

26 July 2020 2,158 4 View

Can anyone explain why do we get diffused bands in western blot? and any trouble shooting techniques?

Hello, Everyone. I have some questions related to western blot as I am new to western blot. I tried to detect the LC3B, however I got diffused bands as I have attached in the file (Please find the...

12 November 2019 8,250 13 View

How much should we add primary antibody into 20 mL of 1% Skim milk for Western Blot?

Hello, Everyone! I currently start doing western blot experiment, but I am not really sure how much should I add primary antibody into 20mL of 1% Skim milk. Here is what I am doing: * I make 1%...

10 November 2019 5,213 9 View

Can anyone explain why do we get diffused bands in western blot? and any trouble shooting techniques?

Hello, Everyone. I have some questions related to western blot as I am new to western blot. I tried to detect the LC3B, however I got diffused bands as I have attached in the file (Please find the...

10 November 2019 5,796 13 View