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

How select composition for ods-high entropy alloys?

31 July 2024 2,795 1 View

Can you suggest any software to draw shadow mask pattern for lithography?

I need some mask for UV photolithography. Can you suggest any software to draw those patterns with proper file format so that directly the file can be inserted and required pattern can be...

22 May 2024 9,449 3 View

How is to find the correct bandgap from a Tauc plot having 2 humps extracted from a UV absorption data ?

In order to find the band gap, I plotted a tauc plot of a UV absorption data. The Tauc plot contains two slopes as shown in the figure. Kindly tell me how to obtain the correct optical band gap...

12 May 2024 5,449 3 View

How can I perform constrained SDFT of a material e.g fcc Ni? Can someone tell me what syntax should I give in my input files?

Recently I am studying on Exchange interaction and Magnetic force theorem(MFT) .. there the basic idea of MFT is we have to rotate the exchange-correlational (xc) potential by assuming that within...

29 April 2024 8,568 0 View

High mRNA expression versus low protein expression?

I tested gene expression by RT PCR followed by Western blotting to test protein expression. I get an inverse correlation with up-regulation at mRNA level and down-regulation at the protein level....

05 April 2024 4,679 6 View

How to find the density of polycrystalline samples?

Hi everyone, I'm working with some polycrystalline oxide sample whose density is not known theoretically. I don't know the crystal structure either. However, I do have the lattice parameter from...

12 February 2024 2,141 0 View

Which Similarity technique is suitable for textual data analysis ?

Cosine similarity, Soft Cosine similarity or SBERT?

02 February 2024 8,149 6 View

What is the standard procedure to optimise the growth of a new materials mainly TMDCs in CVD technique?

I want to grow TMDCs like ReSe2 in CVD ( dual zone either APCVD or LPCVD). How is to optimise the growth for a monolayer ReSe2?

23 December 2023 7,829 3 View

Could you suggest me any tool for adapter trimming of Pacbio SMRT dataset?

I am currently working with PacBio SMRT sequencing data acquired from various sequencer platforms, including Sequel I, Sequel II, and PacBio RS II. These platforms generate both Circular Consensus...

05 December 2023 8,309 0 View

How to plot Effective permittivity and permeability variation with respect to frequency for FSS unit by using HFSS Software ?

The sample graph is attached please anyone explain how to plot the attached graph in HFSS software.

03 December 2023 8,122 1 View

Does anyone know what might be causing the smeared bands on my western blot?

I got these smeared bands quite often lately. We typically run the gel at 140V with a 10-12% gel and do a wet transfer at 220 mA for 1.5 hr in cold room. We also noticed some dirty spots/dots (see...

10 August 2024 7,480 3 View

AMAXA Lonza nucleofection lower volume of buffer?

Does anyone tried to do nucleofection with AMAXA by Lonza with lower than recommended amount of buffer in the cuvettes (100 ul)?

07 August 2024 4,616 0 View

Why I can't see any band in SDS-PAGE?

Currently, when I run SDS-PAGE, I don't see any bands at all, even though I used the same material just a day ago and it worked fine.... In our lab, we dilute the 10X running buffer to 1X and...

06 August 2024 5,373 2 View

Why does my protein refolded to beta sheet during thermal denaturation analysis?

Hi! So i attempted to understand a novel protein behavior towards heat application by analyzing its secondary structure change. I subjected the protein to a thermal denaturation analysis using...

06 August 2024 1,989 3 View

Why is my ASO degraded quickly on agarose gel?

I have been working on Red blood cell-derived extracellular vesicles as Antisense Oligonucleotide (ASO) carriers. We normally run agarose gel to quantify the loading efficiency. I used naked ASO...

06 August 2024 3,130 2 View

Why might the impedance values for DI water and 0.1X PBS buffer solution exhibit a decreasing and increasing trend, respectively over time (HP 4194A)?

Hello everyone, I'm encountering an issue with my electrochemical impedance spectroscopy (EIS) measurements and would appreciate some insights. Experimental Setup: Electrodes: Gold interdigitated...

05 August 2024 3,783 2 View

Low-yield gel extraction problem?

I am having an issue with my gel image where my PCR product is not appearing very bright on the gel. When I perform gel extraction, the A260/280 purity value is very low. I used the Qiagen gel...

05 August 2024 9,798 3 View

What is the best blank for nanodrop if I want to read a recombinant protein concentration?

Is it the "elution buffer" or the "dialysis buffer"? Note: I'll be using NanoDrop OneC

01 August 2024 967 3 View

Transfection in HEK293T cells?

Dear All, I am trying to transfect a pCDNA3.1 vector containing my gene of interest. The purpose is to figure out the localization of the protein of interest. I have fused the protein with GFP on...

31 July 2024 9,892 4 View

PCR showing no bands - Master mix and primers didn't mix?

Hello everyone, I performed a PCR yesterday, and the results showed no bands on the gel. Of course, I probably missed some crucial steps, like adding my samples to the PCR strips themselves, for...

31 July 2024 2,406 6 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?