Selectively visualize his-tag fusion proteins

More Saaraj Gupta's questions See All

What are the most reliable approaches to extract key continuous frames from a video, with complete automation?

I have an input video, of skin (from a microscope that means it is magnified such that blood capillaries are visible). The idea is to extract 15 sec of continuous frames from a video of 1 min,...

02 March 2021 2,735 2 View

Which microtomy technique do you suggest for thin and delicate root sectioning ?And, how to preserve and process the roots before sectioning?

We have thin roots of from 1 year old different species of plants. We are looking into anatomical details of the roots. What's the best method to preserve and process the roots once they are...

23 February 2021 1,898 1 View

Has anyone done Archard Wear Simulation of GFRP Composite Plate in ANSYS with ACT extension or UDF?

I'm looking to do a sliding wear (plate-on-plate) simulation of GFRP composite plates with Archard Wear Model. Any hints or example steps are appreciated.

17 February 2021 1,447 2 View

Which root in this tap root system, in the picture attached, should be used for root sectioning in order to study the root anatomical details?

There are numerous roots in this tap root system. Kindly suggest or share a research paper where I can clarify the doubts as to which root should be chosen for sectioning and for further root...

14 February 2021 1,816 3 View

Possible problem for showing 0.2 sun intensity instead of 1 sun intensity in AAA class Newport solar simulator?

We have a AAA class Newport solar simulator. We changed the Xenon Arc Lamp recently (XBO450W, Make:OSRAM) as the lifetime got over. However, the intensity is coming to be only 0.2 sun instead of 1...

14 February 2021 9,998 3 View

How can I get access of UNAfold tool?

I am searching for tools to construct molecular beacons which shall be complementary to a given sequence. I don't want to design any primers. I found that UNAfold is one such tool which I could...

02 February 2021 9,065 5 View

The peptide synthesized with CTC resin have mass more (+68 Da) than the desired mass of the synthesized peptide. What could be the reason?

I am synthesizing 7 amino acid long peptide sequence with CTC resin, 50oC coupling for 10 min. On cleavage (TFA/TIPS/Water 95:2.5:2.5) I got peptide with good yield. However, It has 68 Da more...

30 January 2021 8,483 3 View

How do we calculate curvature value in ArcGIS?

Hello everyone, I am trying to prepare curvature map in ArcGIS 10.4, using spatial analyst tool. I am using 10 m CartoDem (projected in UTM 44N). I am getting very different value, curvature...

12 January 2021 504 4 View

How can a publish my case report?

I want to publish a case report regarding jejunal diverticula

02 January 2021 7,630 5 View

How can we maintain the two different DC high voltages of same polarity between parallel electrodes inside the vacuum separated by few millimeter?

I am trying to maintain two different high DC voltages of same polarity (either positive or negative) between two parallel electrodes separated by 500 um inside the vacuum. However, what I found...

21 December 2020 1,735 3 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

Will a 10xHis-tag prevent anti-MBP from binding to MBP?

I have a protein with a 10xHis-MBP tag on the N-terminus. I'm fairly certain I am expressing my protein, but I'm not getting a signal on my western blot. Is there a possibility that the 10xHis tag...

25 February 2021 8,404 3 View

How to replicate the well conditions of a crystal hit with 20%(w/v) isopropanol?

I had a crystal hit in a well with 0.1M HEPEs pH 7.5, 10%(w/v) PEG 4000, and 20%(w/v) isopropanol. Why is isopropanol in w/v here if it's just solvent? How do I recreate this well condition?

25 February 2021 9,761 1 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 is the best way to manage Western blots?

Increasingly, more articles are being retracted because of duplicate or otherwise dubious Western blots. Consequently, journals are asking for original Western blots and authentication of...

23 February 2021 8,609 3 View

How to manage western blot for proteomics data validation?

Hi Everyone, I am a bigginer in western blotting and I am trying to use it for validation of my proteomics data. I have: two regulated proteins with molecular mass of 40 and 73 kDa one control...

23 February 2021 6,671 4 View

How to reduce static electricity of powder after lyophilization?

I am working on synthesizing and then purifying peptides using the SPPS system. After purification of the peptide, the peptide powder is obtained using a freeze dryer. When measuring the powder,...

22 February 2021 10,099 5 View

Diltuions for a Western blot?

I need help using the equation C1V1=C2V2 figuring out how much antibody to add to use for a western blot. The stock was origianlly 3.45 ng/ul but diluted 1:10 so now its 0.345 ng/ul. I need to run...

22 February 2021 9,901 3 View

Can anyone help me how to get the similar bands in Actin?

I run western blot on HCT-116 cell lines, I got actin band like in the attached picture. The band size is not same. Can anyone help me explain this situation and how to overcome? Thank you your...

22 February 2021 6,937 3 View

I see 2 random bands in my ERK/pERK blot? Has anyone else seen this?

I ran a western blot using 12% gel and probing for ERK/pERK (cell signaling antibodies). The green is total ERK, the Red is supposed to be phospho-ERK. I see 2 bands below the total ERK that...

22 February 2021 1,777 3 View

Nor Aziyah MatRahim

Hi Saaraj. I'm not quite sure what do you mean by "quantify (visually)". Your question is not clear. In which platform you would like to view. Is your protein is the extracted protein, OR you would like to visualize your protein in cell culture based system?

If it is the extracted protein, I guess you should run the protein on SDS-PAGE and perform Western Blot. Then, you can stain the membrane using the commercially available Ab. There are software the one that I know is Quantity One (Biorad) may allow you to do the analysis to quantify your protein band by scanning the blot and do the analysis.

Saaraj Gupta

Thanks for your suggestion. Protein will be in cell culture based system when I want to quantify it.

Md Wasim Khan

u can do a western blot and probe with anti his Ab