Western Blot

Identification of proteins or peptides that have been electrophoretically separated by blot transferring from the electrophoresis gel to strips of... | Contact experts in Western Blot to get answers

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Questions related to Western Blot

Does low vapor pressure mean strong intermolecular forces and what kind of liquid will have the highest vapor pressure value?

29 August 2023 9,612 0 View

What is the best statistical procedure for a 4x3 experimental design?

or an experimental effectiveness (or a randomized controlled trial) study in which three intervention groups and a control group undergoing an attentional network test (three networks to be...

18 May 2023 8,780 2 View

Why is seasonality more extreme in Northern Hemisphere & why is there less variety in seasons in southern hemisphere than in Northern Hemisphere?

08 April 2023 6,724 3 View

Any ideas on how to get rid of the blue overexposed marker on WB membrane image scanned by LICOR-Oddesy?

I am using the LI-COR-Odessy system to image the membrane of western blots. Sometimes I can see blue markers covered on my bands. Maybe it is the indicator of overexposed bands. Does anyone know...

14 March 2023 474 3 View

What does it mean by 'Iteration limit reached. The results that follow are therefore incorrect' in AMOS??

I'm running a CFA Bifactor model to validate a translated questionnaire (using AMOS). The AMOS output showed 'Iteration limit reached. The results that follow are therefore incorrect'. What does...

11 October 2022 1,100 2 View

Double strand breaks (DSB) and ATAC-Seq?

Is it possible to predict double strand breaks (DSB) (or repair??) using only ATAC data?

08 October 2021 4,218 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...

25 February 2021 8,770 3 View

Has anyone experienced a double band GAPDH in Western Blot?

I am using my GAPDH as a housekeeping protein for my Western Blot. For all the blots I have, my GAPDH was coming out as two bands. Therefore, I am using both bands for my analysis. Has anyone...

12 February 2021 6,685 3 View

Western Blot Total Protein Transfer?

Im new to Western Blots. Im currently working under the analysis part in Image lab. I watched a couple of videos on how to use it and it says to upload two images (Total Protein Transfer and ECL)....

04 February 2021 2,146 3 View

Does Anyone Know of a Way to Detect Melatonin that is NOT HPLC, RIA or Mass Spec?

I tried to do a Slot Blot and was not successful and melatonin is only .23 Kda so it would probably run off a western blot. Has anyone had success with other methods?

24 January 2021 1,590 3 View

How to see the expression of 6kda protein?

Dear all, I am now investigating the expression of small size protein (6 kda). However, the expression of the protein cannot be shown clearly after performing western blot. Therefore, are there...

14 January 2021 3,332 3 View

Why are my western blot bands are think and faint?

Hi, my western blot bands are appearing at the correct molecular weight but are very thin and faint, making it difficult to distinguish difference in expression for shrna samples. Conditions...

05 January 2021 9,386 4 View

Minimal protein concentration from BCA can be detected using western blot?

hello, i extracted protein from PBMC's and after applying BCA assay on 2 samples the protein concentration was 400 ug/ml which means 0.4 ug/ul so does western blot work with these quantities? any...

25 December 2020 6,760 3 View

Is it possible that the phosphorylated proteins mobilize faster than the non-phosphorylated ones?

The band downshifts on Western-blot, after kinase assay treatment.

24 December 2020 8,143 3 View

How to optimize voltage and ampere for transfer of protein to PVDF?

Hello, I looked for a principle for the transfer procedure. I know that it depends on your instrument, buffer, Mw of proteins and the size of paper. I prefer to use constant ampere due to the...

22 December 2020 7,616 3 View

Unspecific band on western blot, what could it be?

I have a problem with unspecific bands on my blot. I have HEK cells transfected with a GFP construct (lane1), mCherry construct (lane2) and RFP construct (lane3). I collect the cells in PBS, do...

18 December 2020 9,417 3 View

Why do I get weird histone bands with my Hep G2 cells??

I'm performing standard histone extraction (abcam protocol) and western blot essay on my HepG2 cells but the band's that I get don't seem to have the classic histone shape. Any clue guys?

15 December 2020 1,839 3 View

How to avoid non specific bands in western-blots..?

I am working on autophagy.. trying to check the release of free GFP after autophagy by western blots.. but i am getting non specific bands.. how to avoid them and develop clean and neat blots ....?

15 December 2020 7,534 8 View

Blue native page transfer problem?

Hello, I am writing this hoping someone can help me. I am just starting out with BN-PAGE and I have a transfer problem. It seems each time I transfer (2 membranes at a time) in a BioRad wet...

10 December 2020 1,217 3 View

What is the difference between actin and loading control in normalization of western blot???

Regarding normalization of western blot analysis, I have found different strategies people follow but I couldn't understand the principle and which one is the accurate, 1- some refer to loading...

05 December 2020 5,903 8 View

Why is hard to produce antibodies that are specific to a protein?

Some antibodies reveal nonspecific bands when a western blot is performed. Why is hard to find a good antibody for a protein? Antibodies produced using as the immunogen a synthetic peptide or a...

04 December 2020 386 3 View

If we forget to add mercaptoethanol, can we correct the samples somehow?

Hi everyone, Can someone help me with a western blotting troubleshooting? So during sample prep, we boil the samples with 2-mercaptoethanol in Laemli buffer. However by mistake, some of the...

03 December 2020 7,095 3 View

What does it mean if you have two variables strongly correlated but only one variable changed in the experiment?

Hello I have two variables measured: one is a behavioural variable and one a western blot variable of a protein. They are strongly correlated. However, only the behavioural variable was...

03 December 2020 1,078 3 View

Why the western blot of HIF1alpha of the whole cell lysate shows intense band under normoxia without any induction of hypoxia?

I am working with oral cancer cell lines and trying to find the expressional changes of HIF1alpha under different experimental treatment. I am getting intense band of HIF1alpha in the western blot...

30 November 2020 5,901 3 View

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