To determine protein sequence?

01 January 2015 0 9K Report

I want to sequence an unknown protein. From where I can get this facility?

Badges
Science topic

More Bhagya Deepachandi's questions See All

Non-specific bands in PCR?

Usually I am doing PCR with 0.5 ul of Taq polymerase (abm) per reaction. Since I wanted to increase the sensitivity of target amplification, I tried PCR with 1.0 ul of Taq. It gave some bands in...

02 March 2018 9,197 6 View

Mass spectroscopic analysis for protein identification?

I am doing immunoprecipitation for concentrating by target proteins. I usually elute my proteins in SDS loading buffer containing Tris, glycerol, SDS, beta mercaptoethanol and bromophenol blue....

11 December 2017 1,246 2 View

Human Anti-Leishmania IgG ELISA Kits...

I have found two commercial ELISA kits for detection of human leishmaniasis. 1. Human Anti-Leishmania IgG ELISA Kit (ab108749) by Abcam 2. Leishmania IgG ELISA Kit (KA3295) by Abnova Both are...

03 April 2017 3,701 1 View

Why did the protein sample and marker not migrate properly in the separating gel (SDS-PAGE)?

I am facing the a problem in my SDS page gel. protein marker and samples not get well separated. All samples were stop separating at the middle of separating gel (protein marker also). Dye front...

09 October 2016 3,411 12 View

Physical characterization of crude parasite Ag???

Is there a simple method to physically characterize the crude Ag extracted from a parasite to check whether they are proteins, polysaccharides or phospholipids like compounds? I am using crude Ag...

07 August 2016 4,034 3 View

Analysis of proteins using westernblotting?

Does 3D structure of a protein affect for migrating distance of a protein?

06 July 2016 6,313 0 View

What is the best condition for blocking western blot membrane?

Can I keep my blocking buffer (skimmed milk or BSA) at room temperature for overnight? Is that affect for my protein?

05 June 2016 3,976 5 View

Commercial protein sequencing services?

I want to sequence some unknown protein excised from 1D-SDS-gel. Have anybody experienced with protein sequencing? Are there any good commercial protein sequencing services?

04 May 2016 4,567 1 View

Fuzzy bands on SDS-PAGE?

What are the reasons for getting fuzzy bands on SDS-polyacrylamide gel?

03 April 2016 594 9 View

TMB SUBSTRATE FOR WESTERN BLOT???

How can we convert TMB of ELISA for western blot by converting soluble TMB product of ELISA into insoluble with addition of 0.05% sodium nitroprusside? Can I use ELISA TMB substrate for Western blot?

02 March 2016 8,684 0 View

Can I base on reverse DNA sequences to perform alignment, convert to amino acids and GenBank submission?

I have reverse sequences (AB1 format), can I base on reverse DNA sequences to perform nucleotide alignment, convert nucleotides to amino acids and deposit the sequence in GenBank database?

11 August 2024 5,138 1 View

How to confirm the site-directed mutagenesis result without performing NGS?

I'm cloning a fragment of 3200 nts into plasmid. The cloning was successful, however, 02 amino acids were mutated. Now I want to fix these 02 aa by site-directed mutagenesis technique using...

08 August 2024 4,645 2 View

Can I use a HisTRAP column for affinity chromatography?

I'm working on selecting antibodies against a recombinant protein that has a His-tag. My idea is to first bind the recombinant protein to a HisTRAP column and then use this column for an affinity...

07 August 2024 505 3 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

Does anyone have issues using Prepman Ultra reagent for MicroSeq ID bacterial, fungal and yeast sample preparation?

I have been attempting to extract DNA from Bacterial, Fungal and Yeast banked samples (>1e7 cells) using Prepman Ultra reagent and I seem to be struggling to obtain a sequence. Although the...

01 August 2024 2,079 0 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

How to retain the a GFP tagged gene expression in stable cell line?

Hello , I established a stable cell line expressing GFP tagged to a centrosomal gene having G418 drug selection marker. I validated the stable line by IFA and Western blotting, results are fine....

29 July 2024 5,007 0 View

What is the relationship between protein structure and N or C terminal tagging choosing?

I want to do 2,3-butanediol dehydrogenase(BDH) enzyme purification to confirm its activity for 2,3-butanediol. Before that, I need to confirm which N or C terminal tagging is better for enzyme...

28 July 2024 366 3 View

Why cannot i find my protein on cell surface after antibiotic selection of expressing plasmid?

I cannot confirm cell surface protein expression by flow cytometry even after transfection and antibiotic selection of cells. Does it take long for proteins to express on the cell surface? the...

28 July 2024 3,178 3 View