Protein extracts not dissovled in buffer?

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Any suggestions on the effect of 1mM sodium citrate solution on reverse transcritpion?

I started using "RNA storage solution (1mM sodium citrate) from THermoFisher to dissolve RNA pellets and store in -20 or -80. However, there is no amplification in the Q-PCR. it is frustrating and...

10 November 2019 5,088 3 View

RNA Quality and formaldahyde Agarose gel?

I am trying to run RNA on 1% agarose/formaldehyde gel using 5x MOP buffer and SybrSafe staining instead of ethidium bromide. what is the proper voltage and time? it says 5-6V/cm, but how can I...

09 October 2019 8,319 6 View

Concentration calculation in multi-analyte ELISA?

Hello, I did multi-analyte ELISA to detect the concentrations of several cytokines, but how should I calculate the concentration as each well represents a different sample and a different...

06 July 2019 4,991 2 View

How long is the 1:1 PCR primer mix valid for?

Does primer mix expire?

05 June 2018 2,846 0 View

RNA Quality and formaldahyde Agarose gel?

I am trying to run RNA on 1% agarose/formaldehyde gel using 5x MOP buffer and SybrSafe staining instead of ethidium bromide. what is the proper voltage and time? it says 5-6V/cm, but how can I...

01 January 1970 5,937 3 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 to increase a model accuracy ?

dear community, my model is based feature extraction from non stationary signals using discrete Wavelet Transform and then using statistical features then machine learning classifiers in order to...

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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,...

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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

How can extract ciprofloxacin hydrochloride disolve in water er to organic solvent such as dichloromethan or any other solvent?

I need a procedure to extract the ciprofloxacin hydrochloride from aqueous solution by using dichloromethane in detail,. I thank you very much.

01 March 2021 4,306 1 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

Which bacteria are suitable for quorum sensing screening?

I want to find a quorum sensing inhibitor from plant extracts. Klebsiella pneumoniae, Moraxella catarrhalis, Staphylococcus aureus, and Streptococcus pneumoniae. I looked for these strains...

01 March 2021 7,675 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

Jai Ghosh

I would suspect that your protein might have got dissolved in the homogenizing buffer and what you are observing as insoluble may be something else. Try to check and confirm.

Jasim Al-saadi

To dissolve your protein try to add urea or SDS to the buffer

Kamelia Mirdamadi

Jasim Al-saadi Thank you, is it 1% or 0.1% SDS?

Kamelia Mirdamadi 1% .

Thank you