Should I show WB bands of protein's full-length and cleavage in single blot image (saturated full-length) or in each seprated images (unsaturated)?

Dear All, I have question on choosing WB bands image for publication. My target protein has two forms: 1.full-length form (very strong signal) 2. cleaved form (faint/low signal) in the same blot....

04 May 2016 7,202 5 View

How can I optimize blush-like/blur? Immunofluorescence (IF) image quality to be more dot-like?

I have done IF with Abcam antibody, but the image that I get is not look like as the Abcam review on the same anitbody. My image (attached red image) is kind blush-like or blur color?, but from...

04 May 2016 8,103 3 View

Is there a way double-color immunofluorescence staining for two different protein, but have same rabbit host species antibodies?

Dear All, I tried to study co-localization of two different proteins by using immunofluorescence staining, since the western blot result of both protein shows significantly increase in nuclear...

03 April 2016 1,461 10 View

Problem with cell deteaced/morphological changed after fixation with 4%PFA, How should I optimize the protocol?

Dear All, I have SH-SY5Y grown on coverslip (precoated with 0.01mg/ml for 1hr at 37C) in 24 well plate. When the cells were ready, I wash it breifly with PBS, then fix it with 4% Paraformaldehyde...

02 March 2016 7,594 18 View

Why FasL protein expresses in western blot lower in H2O2-induced cell death (compare to Ctrl)?

Dear All, I am study FasL protein expression using western blot in oxidative stress-induced cell death in SH-SY5Y. My results mostly shows that FasL expression is non-significant changed or...

02 March 2016 8,956 4 View

What is different between RM or Ordinary one-way ANOVA & What kind of experiment is suitable for each of them?

Dear All, I just switch from SPSS to Graphpad prism 6. In analisys, there is option to select experimental design between RM (repeated measure) and Ordinary (no pairing) for one-way ANOVA. What is...

01 February 2016 3,821 4 View

Is Fas ligand's western blot signal strong in general cell line?

01 February 2016 7,443 2 View

How should I select western blot band to measure if getting multiple target protein bands? any online database to look for?

Dear All, I am real newbie in protein field. I would like to ask you a few question regarding to select band to measure in western blot. In case that blocking is fine and I use single primary...

01 February 2016 1,304 8 View

How should I solve blochy/uneven background problem on western blot?

Dear All, In my western blot experiment, I have problem with blotchy blackground (like in attachecd picture) on the blot, which usaully appear when I expose low signal band for 15-30 min or more...

31 December 2015 3,101 7 View

How can I analyze a western blot to determine significant difference of a target protein that has no band in Ctrl, but shows band in condition group?

Dear All, I have a problem to do stat analysis to determine significant difference between control group and each condition group (vary dose of H2O2 treatment to cell line) of a target protein in...

11 December 2015 4,456 13 View

Protein-aptamer gel electrophoresis help, please??

Hi, I have problems with running gel electrophoresis. I have tried agarose gel electrophoresis and native PAGE. I have two proteins, which have molecular weights of ~30kDa and ~180kDa and two...

03 March 2021 4,275 4 View

Gel electrophoresis result for total RNA samples, 1.5 Agarose gel was used, How to improve the integrity of RNA?

Gel electrophoresis, RNA degradation, RNA extraction from fresh tissue

02 March 2021 5,433 5 View

How to regulate SSCP temperature in simple PAGE?

I'd like to perform single-strand conformation polymorphism (SSCP) in my thesis, however I cannot control the temperature of the vertical PAGE since we are using the conventional tanks. Is there a...

02 March 2021 9,157 1 View

Why are my bands weird in this Agarose gel?

Can someone please give me some possible things that could go wrong? Here is my recipe: 0.5g Agarose 50 mL of TAE 1x 1 uL ethyl bromide. Gel was run at 100V for 1 hour. The buffer used is also TAE.

01 March 2021 9,952 3 View

Which fluorescent dye to use for measuring total ROS production?

Is DCFDA appropriate for measuring total ROS production, or can it only detect certain types of ROS? If I want to measure total ROS production, which dye should I use?

01 March 2021 9,447 3 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

How can I represent SDS-PAGE results for machine learning analysis?

I am trying to classify and analyze the results of an SDS-PAGE based array for bacterial detection using machine learning, but I have trouble finding the best way to represent the results with...

27 February 2021 9,176 3 View

Is there anyone who experienced with cytotoxic effect of Hoescht 33342 dye?

Is Hoechst toxic to cells? In general, it is known as non-toxic. I am working with dsRED MDA-MB-231 cell line. I applied it for my PDMS-based microfluidic platform, for this reason, I incubated...

26 February 2021 9,119 3 View

Why my agarose gel staining sometimes looks not homogenic but like gradient (see the figure)?

I am worried about this overexposure of the upper part of the gel in the picture. Is it possible that this is the result of too much concentration of ethidium bromide? Why is there such a big...

25 February 2021 8,140 3 View

EMSA troubleshooting. Protein-DNA complex not migrating, cold probe lane with multiple bands. Can anyone help?

I'm running an EMSA to show the DNA binding activity of recombinant proteins versus the Wild type. Previous assays run by my research group using a different test system show the protein-DNA...

24 February 2021 8,325 1 View