Does denaturing conditions alter the protein fluorescence?

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Is there any particular site to insert the fragment in histag vector for further protein purification?

Do we need to place the insert in a specific site in his tagg vector for further protein purification or can we just use any restriction site present in the vector?

08 September 2019 8,351 3 View

Why do i find another protein size on my gel?

I've cloned a gene which size was around 800bp in pet 32 vector next to n-terminal histagg to further purify the protein. when I expressed the protein there was a big band around 50 kda + my...

08 September 2019 6,597 5 View

Why has the protein loss it's red color?

i'm doing a protein purification that i incorporated with mcherry because i want to visualize that after purification.i've tried low temperature 14 degree as well as 37 degree but the protein and...

08 September 2019 3,788 4 View

Wher can be the issue if the cloning doesn't work?

I"ve been doing a cloning about 4 months already. i've done all the controls and I 've used differents vectors and also I've done vector dephosphorylation though I used double restrictions...

05 June 2019 4,343 12 View

Does anyone know a vector which can express protein easily?

does anyone know an histagg vector that I can use to clone a gene easily and further express my gene protein ?

05 June 2019 9,282 3 View

Is there any diifference between self ligated vector and fragment-vector ligated?

after we have done transformation ,is it possible to compare colonies on plates, i mean is it possible to find that fragment-vector ligated are bigger than self-vector ligated?

03 April 2019 2,695 3 View

What is the importance of iptg for protein expression?

sometimes the protein can be expressed even without adding iptg, so is it necessarily to add iptg for protein expression?

03 April 2019 7,650 2 View

Is it possible to do the insert digestion in two steps?

I'm trying to clone my insert in pet 16b with nde1 and xho1 restrictions enzymes but I'm always getting only self ligated vector ,and the vector control also have many colonies than the...

03 April 2019 9,455 3 View

how can i clone my gene in pet 16b cloning for further protein purification?

i'm trying to clone my gene in pet16b to further make histagg protein purification. Where can I do the cloning for a successful cloning and which restriction enzymes to use? I also want to get a...

02 March 2019 6,238 6 View

Can i use these enzymes to digest my vector and fragments and use the same to test the ligation efficiency?

my primers are F:GCATCATATGatgaatgggaccgcaaaccc and R:GCATCTCGAGttagtctttctggtaaggccg,my fragment that I want to clone start by atgaatggga.......,and my vector is pet16b.i used ndeI and XhoI...

02 March 2019 4,189 9 View

Hello researchers Is this a random laser or just fluorescence?

I am using Rhodamine6G as gain medium and silver nanoparticles as scatterers on a microscope slide and laser input 532 nm comes from above.

09 August 2024 9,894 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

Weak DAPI staining after immunohistochemistry - how to improve?

After immunohistochemistry of previously fixed in PFA and EtOH and then frozen 20 μm sections of zebrafish brain, DAPI staining is very weak (right) compared to the same sections stained without...

05 August 2024 9,637 2 View

Are there any commercially available Donkey anti-Alpaca secondary antibodies?

Are there any fluorescently labeled anti-Alpaca secondary antibodies raised in Donkey? So far I have only been able to find anti-Alpaca secondaries raised in Goat. Or is this not possible due to...

04 August 2024 4,255 1 View

Why results of ROS flurescence are negative as there was no bacteria within?

Hello. I am working on ROS production of two systems: system A is cerium oxide and hydrogen peroxide, system B is cerium oxide nanoparticle, hydrogen peroxide and potassium bromide. I did some...

04 August 2024 5,974 3 View

How to calculate the molar proportions of the oxides from the XRF analyses?

02 August 2024 2,565 2 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

Is it necessary to attach fluorophore at 3' end in MIC RT PCR machine for Dual hybridization Probe designed especially for genotyping assay?

I designed 2 dual hybridization probes for my single SNP. where I attach my fluorophore 5'-FAM-nnnn-BHQ1-3', and 5'-HEX-nnnnnn-BHQ2-3'. I need to run it on the MIC RT PCR machine by Biomolecular...

28 July 2024 5,658 0 View