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

Enhanced Yellow Fluorescent Protein (Aequorea victoria) DNA sequence?

I am on the lookout for the Enhanced Yellow Fluorescent Protein (Aequorea victoria) DNA sequence. Does anyone know where I can find it? Thank you in advance

03 March 2021 3,568 1 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

Buffer Exchange using column packed with sephadex G-25 effect on protein concentration?

Hello, If i am doing a buffer exchange for an antibody of 1mg/mL, does the elution lose protein in the process of buffer exchange? For example, if i flow through 500 uL of 1mg/mL sample, and...

03 March 2021 6,299 3 View

How optimize the Size Exclusion Chromatography?

Hello! I'll do a size exclusion chromatography, but I only have an open column, and I'll perform the peptide extraction from yeast, using buffer lysis (sodium phosphate 50 mM/NaCl 30 mM/DNAse and...

01 March 2021 2,215 2 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

What do I do with DLS peaks that are from the buffer?

I'm looking at the aggregation of my protein sample using DLS. Unfortunately, my buffer (20mM HEPES) also results in a set of peaks. These are at approximately 1 and 1000 d.nm. The lowest peak...

01 March 2021 9,015 2 View

What is the most suitable resuspension buffer for TCA/acetone precipitated protein?

I need to extract protein from fermented carbon sources for Bradford assay. Most researchers experiencing insolubility of pellet in resuspension buffer. Please assist me to select most suitable...

26 February 2021 7,129 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 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

Cloudy/high background western blot - can anyone help me?

Hi everyone! I try to detect VDR protein (~48 kDa) using Western Blot. After SDS-PAGE using 10% MP TGX Stain-Free Gel from BioRad I transferred the protein onto an metOH activated PVDF membrane...

23 February 2021 6,266 3 View