In one of my Western blot results all my protein bands (33- 57 kDa) appeared at the top (protein marker was fine) 175 to 270 kDa. Reason?

More Debasis Sahu's questions See All

Can anyone suggest papers in the hybrid auto-mata area that models cyber-physical systems in power systems?

Most of the hybrid automata papers target formal verification of the systems. I am looking for papers that are solving optimal response problems that propose optimal control in power systems under...

07 February 2021 5,233 1 View

What is the recommended age limit for primary posterior capsulotomy in pediatric cataract surgery?

In our centre, we are doing PPC even for older children upto 10-12 years of age and there seem to be no harm, rather beneficial and good outcomes. Please share your experiences.

27 December 2020 4,268 6 View

Increase coercivity with device size (multi-domain) ?

I'm working with magnetic material particularly CoFeB. I notice a weird pattern. I see that my coercivity increases with device size. The device is multi domain and contrary to what I'd expect....

09 December 2020 1,577 1 View

Software for L x T mating design analysis and heterosis estimation?

Kindly suggest the name of free softwares available for LxT analysis and heterosis estimation in crop plants.

22 October 2020 1,744 10 View

I have performed XRD on Fusion zone of 316L stainless steel as well as incoloy 800. Why only one or two peaks are showing?

I am facing problem as only one or two peaking are coming while performing XRD on Fusion Zone of 316L SS and alloy 800. I want to know where i am committing mistake.I have attached the XRD image.

07 October 2020 6,205 3 View

Please suggest the name of important and free softwares for attractive NJ tree, dendrogram and PCA diagram?

I would like to do genetic diversity analysis. Kindly suggest the name of important and free softwares for attractive NJ tree, dendrogram and PCA diagram?

30 September 2020 4,684 7 View

Band decomposed charge density using quantum espresso ?

Dear sir/mam Greetings for the day ! (1). I am using quantum espresso(QE) to calculate band decomposed charge density. To my knowledge, the kpoint at which we want to calculate the charge...

21 August 2020 1,095 2 View

How to find out the Compund Average Annual Publications Growth Rate (CAGR) in Bibliometrics ?

I need an explanation with example and calculation.

30 July 2020 8,765 1 View

What is the best graph based database for storing complex graphs from diverse domain?

This question is related to making inferences from a complex graphical model. Graph databases are used to store and query the relationship between nodes in a graph. We need an open-source graph DB...

18 June 2020 5,903 3 View

Is rationality normative? Can rationality be naturalized? Is rationality depended on normative conditions, or is it independent of normativity?

Rationality is itself an elusive term. There are debates on the definition and criteria of rationality. The primary assumption of the naturalized and non-naturalized rationality on normative...

05 May 2020 1,095 78 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

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

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

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

Saravanan Ramanujam

Try again with freshly prepared loading buffer in 8 to 10% of gel. And try to include a positive control sample if possible.

Omar Santín Martínez

Add DTT to your protein purification buffers since sometimes disulfure bonds could be the answer!!

Emiliano Melgar-Bermudez

I agree with Omar, I don't think the proteins were denatured and there may have been some aggregates as well.

Mohammed Arif

use fresh SDS buffer with BME. and boil the sample for 5-15 min

Shashank Shekhar

Hi sir,

What sample you are using here?