Why does increasing extracellular pH increase the proton motive force in bacteria?

I thought increasing proton concentration (lowering pH) would increase the proton motive force, but I'm reading that the opposite occurs. Trying to understand why increasing pH increases...

11 December 2019 4,549 3 View

Can anyone help me predict if the fluorescence spectrum of NBD-CO-Hz will change in this reaction?

I am reacting NBD-CO-Hz with oxalate. When reacted (condensed) with a single carboxylic acid-containing molecule the fluorescence spectrum of NBD-CO-Hz does not normally change much. It is...

08 September 2019 1,380 1 View

Why is PDAM Giving me this Result on UPLC?

I have purchased PDAM from two different suppliers, and am getting the same problem both times. This chemical should be non-fluorescent until reacted with a carboxylic acid, but when we run the...

06 July 2019 6,454 3 View

PDAM (1-Pyrenyldiazomethane) fluorescence derivatization of oxalate?

Hello, I have a question about PDAM that I hope someone can help me with. I would like to measure oxalate (a dicarboxylic acid) in aqueous samples, and I need to find a suitable fluorescent...

31 December 2018 8,893 4 View

Do CHO cells polarize on Transwell inserts?

CHO cells are reportedly of epithelial origin. I have a transporter stably transfected in the CHO cells that is normally apical in the colon. Will it be exclusively expressed on the top aspect...

06 July 2018 328 2 View

Incubating cells in 50 mM bicarbonate, pH 7.4, why does intracellular pH go down?

I am performing an experiment in CHO cells with or without a chloride/bicarbonate exchanger protein expressed in the membrane. When I replace normal buffer containing 130 mM chloride (pH 7.4)...

05 June 2018 524 2 View

How long can CHO cells function without magnesium?

I am studying transport of something that can bind to and be inhibited by magnesium, so I am excluding magnesium from the transport buffer. I find that after washing the cells and incubating...

03 April 2018 5,509 3 View

What is the best way to irreversibly protect or modify the nitrogens in urate to make them tertiary? I need to accomplish this in aqueous?

What is the best way to irreversibly protect or modify the nitrogens in urate to make them tertiary? I need to accomplish this in aqueous conditions. I am trying to modify them to avoid a...

02 March 2018 222 0 View

Does phenyl isothiocyanate react with the amines in uric acid?

PITC reacts with ammonia, primary, and secondary amines. Therefore, it should react with uric acid, correct?

10 November 2017 6,603 0 View

In a suspension of Ca oxalate is it possible to pull the Ca from oxalate and force it to bind to carbonate? If I supply abundant Na bicarbonate?

04 May 2017 2,739 2 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 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

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

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

What is the most suitable resuspension buffer for TCA/acetone precipitated protein?

I need to extract protein from fermented carbon sources for Bradford assay. Most researchers experiencing insolubility of pellet in resuspension buffer. Please assist me to select most suitable...

26 February 2021 7,129 3 View

What urea buffer can i use for biotagged protein extraction from streptavidin beads?

Hi all, I have been doing research on biotagged LDB1 proteins in Mel cells. I purify the proteins using M280 streptavidin beads. The yield is very low so I've been trying to optimize it by...

24 February 2021 5,749 3 View

Cloudy/high background western blot - can anyone help me?

Hi everyone! I try to detect VDR protein (~48 kDa) using Western Blot. After SDS-PAGE using 10% MP TGX Stain-Free Gel from BioRad I transferred the protein onto an metOH activated PVDF membrane...

23 February 2021 6,266 3 View

How to improve the buffer capacity of sea water?

Hello everyone I would like to improve the buffer capacity of seawater (pH ~8) in order to dissolve as much CO2 as possible without dropping the pH below 7.5 (a range between 7 - 7.5 is...

17 February 2021 2,797 2 View

Why does flow deformation occur at the test line and control line on nitrocellulose membranes (LFIA)?

I use 1% BSA in 10 mM phosphate buffer (pH 7.4) solution when striping my nitrocellulose membranes with antigens and antibodies. However, I always observe deformed flow when my sample or running...

16 February 2021 4,224 3 View