Transwell migration assay - cells dying !?

Hi all, using a transwell assay to test migratory function of cytokines, Costar transwells 5 micrometer, 24 well, according to publications and protocols 100 ul in top compartment with 1x10*6...

09 October 2011 4,678 2 View

MSP stadardization problem

Read 1 answer by scientists to the question asked by Axel Hueber on Oct 8, 2009

09 October 2009 6,627 0 View

protein science

Read 2 answers by scientists to the question asked by Axel Hueber on Oct 8, 2009

09 October 2009 7,156 1 View

Caspase 3 - Please Help!

Read 4 answers by scientists to the question asked by Axel Hueber on Sep 18, 2009

08 September 2009 1,153 2 View

nonspecific bands in CO-IP

Read 2 answers by scientists with 1 recommendation from their colleagues to the question asked by Axel Hueber on Sep 18, 2009

08 September 2009 6,525 0 View

SINGALING PATHWAY IN A CELL

Read 1 answer by scientists with 1 recommendation from their colleagues to the question asked by Axel Hueber on Aug 1, 2009

07 August 2009 3,593 0 View

i need help w HMEC culturing

i'm culturing my HMEC in gelatin -coated flasks (by me) using MCDB medium and EGF, heparin and FBS (10%) and the cells did not attach... what...

07 August 2009 6,047 0 View

copy number determination: sauthern blott vs real time pcr

Read 2 answers by scientists to the question asked by Axel Hueber on Aug 25, 2009

07 August 2009 156 0 View

Sequencespace

Read 1 answer by scientists to the question asked by Axel Hueber on Aug 25, 2009

07 August 2009 1,030 0 View

hi

07 August 2009 4,848 2 View

Why might my GFP tag be dissociating from my plasmid after transfection?

I have a set of stably transfected cell lines all transfected with plasmids containing GFP tags on the C terminus. During a western blot using anti-GFP antibody, one of my plasmids has dissociated...

01 March 2021 9,310 4 View

Precipitate after adding laemmli buffer to protein solution containing Potassium Acetate?

Hi, I am running a size exclusion chromatography experiment with a buffer containing Potassium Acetate as a salt. I analyse these fractions through SDS-PAGE. After boiling my SEC fractions in...

01 March 2021 2,622 3 View

What is the most suitable server for predicting protein structures in bacteria?

I have used the i-Tasser several times, however it has been unavailable for several days. I tried the swiss-model, but the output was not very pleasant due to the model used. Is there any other...

28 February 2021 4,521 3 View

Do you need the CMV enhancer before the CMV promotor for a proper function of CMV on the gene that follows?

I'm a student and I have to produce a cell line with a knockin for the NRF2 gene with GFP. I have to put a promotor in front of the GFP because the gene will be too far away from the promotor of...

28 February 2021 7,127 2 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

How to replicate the well conditions of a crystal hit with 20%(w/v) isopropanol?

I had a crystal hit in a well with 0.1M HEPEs pH 7.5, 10%(w/v) PEG 4000, and 20%(w/v) isopropanol. Why is isopropanol in w/v here if it's just solvent? How do I recreate this well condition?

25 February 2021 9,761 1 View

Problem with ligating 2.8 kb insert into 8.5 kb plasmid?

I am trying to clone 2 copies of the same mammalian gene into the pSF-CMV-Ub-Puro Ascl (contains HindIII, KpnI and NheI cut sites) plasmid. This is what the final product is supposed to look like...

24 February 2021 6,310 3 View

What urea buffer can i use for biotagged protein extraction from streptavidin beads?

Hi all, I have been doing research on biotagged LDB1 proteins in Mel cells. I purify the proteins using M280 streptavidin beads. The yield is very low so I've been trying to optimize it by...

24 February 2021 5,749 3 View

Can I use a protein-devoid condition as my negative control in ELISA?

Hello, I am running a sandwich-type ELISA where I am trying to inhibit a protein-protein interaction using small molecules. For each experiment, I determine my Z' (Z-prime) to determine the...

23 February 2021 7,429 1 View

Is there a way to extract total RNA and protein from cell culture at the same time, without denaturing protein?

Hi all, Is there a way to extract RNA and protein from the same cell culture sample? Because we need to use the protein for gelatin zymography at later time, we are looking for a method that can...

22 February 2021 5,341 5 View