Which one is the better plan for time course study?

I am doing a time course experiment. Treat cells with a drug for different times (0h, 2h, 8h, 18h, and 24h, respectively), and then harvest cells for RNA to check cell responses. So here we have 2...

31 December 2016 4,533 9 View

How to stabilize formamide?

How to prevent formamide being oxidized to formic acid? Thank you so much for any ideas.

10 November 2016 2,149 0 View

What any effects of stearic acid treatment on protein concentration?

02 March 2016 4,914 3 View

Do you add M-CSF or L929-conditioned medium to cell culture when you treat BMDM cells?

02 March 2016 9,974 11 View

Does anybody have good idea with three plasmids co-transfection into macrophage cells?

I am trying to co-transfect 3 plasmids into macrophage cell line (RAW264.7). I can transfect single plasmid successfully and can detect with immunoblotting. But it is very hard for me to do...

09 October 2015 5,465 8 View

What 2x Laemmli Sample Buffer recipe is better?

Up till now, there are two kinds of 2x Laemmli sample buffers: Buffer 1) 65.8 mM Tris-HCl, pH 6.8, 2.1% SDS, 26.3% (w/v) glycerol, 0.01% bromophenol blue Please refer to the original...

04 May 2015 1,878 8 View

How do you make acetylation detection easier with Western?

I treated macrophage cells with LPS (1h to 24h time course) and tried to detect p65 acetylation. Method 1) I collected total cell lysate with RIPA buffer containing 10mM NAM and 10uM TSA. And then...

31 December 2014 7,371 8 View

What is the best method or kit to purify mono-antibody from tissue culture?

08 September 2014 3,162 4 View

Can anybody help me with western blot using small intestine tissue lysate?

Here is my protocol: 1. Collect small intestine from mice and push out the stool. 2. Flash freeze in liquid nitrogen and then store the tissue at -80C for later use. 3. Homogenize tissues in RIPA...

02 March 2014 4,307 9 View

Does a forest free freezer really destroy antibodies?

All freezers (-20C) we have are forest free. Is it safe to keep antibodies in those freezers? If safe, how many months?

02 March 2014 426 9 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

Should we change media after1st 6hrs after plating HUVEC bought from INVITROGEN?it's not recomended by invitrogen.Will DMSO present in stock matter?

We have HUVEC bought from Invitrogen. Cat no -C-003-5c. while first plating the cells we have bought the protocol suggests not to centrifuge the cell with media to remove the DMSO. Should we...

02 March 2021 3,713 2 View

Sample dilution for quantitative analysis using ELISA?

If the detection range is in ng/ml but the reference range is in ug/ml for a molecule or protein in serum or plasma .how to dilute and what is the initial volume to be taken for quantitative analysis

02 March 2021 7,670 3 View

Are protein samples in SDS compatible with ELISA?

I have used an AllPrep DNA/RNA/Protein Mini Kit QIAGEN kit to extract RNA and protein from my samples. At the end of the protein purification, I resuspend my protein in 5% SDS. Will these samples...

28 February 2021 7,370 3 View

Does anyone use antinuclear antibody (ANA) and anti-Phospholipid antibody ELISA kit for Non Human Primate serum samples?

I am looking for anti-nuclear antibody (ANA) and anti-Phospholipid antibody ELISA kit for Non Human Primate serum samples. Any suggestions highly appreciated. Thank you.

28 February 2021 6,512 3 View

Anyone know of a good progesterone antibody?

I am looking for a good progesterone antibody to quantify levels of progesterone in zebrafish. Applications would be ELISA, and immunohistochemistry.

28 February 2021 5,929 2 View

Benefits of the 24 welled plate?

We have always used the 96 welled plate, the 48 welled plate but apparently there is much evidence on the 24 welled plate and above all its benefits.It states it is the same you would use on ELISA...

28 February 2021 1,149 1 View

Bought HUVEC from Invitrgen.need to revive it.expected it is cryofreezed in DMSO.Do I need to remove the DMSO for plating the cell for the 1st time?

After I cryofreeze the cell do I need to remove the DMSO for culturing the cell for next passages .

28 February 2021 7,738 3 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