Does imidazole influence in flow cytometry for antibody fragments?

15 September 2022 0 5K Report

I am screening several scFv sequences by flow cytometry. I got good expression/purification yields and next step is dialysis to remove imidazole and then flow cytometry to see if they bind to expected target. However, I wonder if it is really necessary to remove imidazole to incubate with cells. Has anyone tested that?

Badges
Science method

More Marcus Bezerra's questions See All

Can holes for pressure probes in journal bearings modify the original pressure distribution?

It is very common to measure the pressure distribution inside the bearings with pressure probes connected to the oil film through holes in the bush. Once filled and pressurized, the probe lines...

12 June 2024 1,936 0 View

Why does the Barricor mechanical separator heparinized plasma tube showed a positive bias for LDH?

The subject of the study centers on the switch from gel separatory tubes to lithium heparin gel tubes in the clinical laboratory but the study additionally addressed the use of the Barricor...

05 February 2024 5,445 1 View

Is it possible to use the parameters of N719 Dye (such as CB and VB electron density) for my natural dye in SCAPS 1D even though I have different dye?

I am using a natural dye (strawberry, carrots, blueberry, turmeri, spinach , arabica robusca). I have the UV-VIs spectra of each. I can calculate the bandgap energy and HOMO and LUMO levels but I...

08 January 2024 3,965 0 View

I would like to konw if there is a alternative to DJI Terra to pre-process LiDAR data from the Zenmuse L1?

LiDAR data collection and processing

07 December 2023 3,745 0 View

After macerating a plant, how do I concentrate the extracts that the yield would be in solid form, it is neither sticky nor liquid?

Kept seeing studies but the results of their extraction is sticky substances. I do stumble upon studies with vacuuming or water bathing the extracts to dryness, but I am skeptical whether the...

11 November 2023 202 0 View

Does anyone know of a psychometric scale concerning perception of HIV Pre-exposure prophylaxis (PrEP) and evaluation of risky sexual behaviour (RSB)?

I'm looking for a psychometric regarding perception of PrEP and sexual expectancies that have items that also include attitudes/perception of risky sexual behaviour (RSB), i.e., condoles sex,...

24 August 2023 4,028 0 View

Does anyone please know of any measures for perception/attitude to risky sexual behaviour (RSB) that would be appropriate for a diverse demographic?

I need a measure for RSB that I can use for a survey that would encompass a wide and diverse population demographic without restrictions to age, gender, race, sexuality, religion, and lifestyles....

24 August 2023 8,098 2 View

How can I compare different Likert scales of the same population over time?

Dear community, we work on a employee satisfaction (done via questionnaire) project in an organisational setting. For the previous years, the questionnaire hat a 5 point Likert scale for all...

31 March 2023 3,525 5 View

Am interested in High Definition Mapping for driverless cars please advise on available material?

HD Mapping of Driverless Cars

06 March 2023 8,840 2 View

What is the optimal Celsius, Millitorr, and Time to Freeze dry a frozen a aqueous plant extract?

I want to froze an aqueous plant extract at -20 °C for 24 hours, then freeze dry it. I want to know if it's okay to freeze an aqueous extract to -20 °C for 24 hours then freeze dry it when I want...

25 February 2023 8,416 2 View

Mouse CD3 antibody sequence?

Can anyone share any reference that gives mouse specific CD3 antibody sequence. I have been searching for this since few days and no luck. Thank you very much

08 August 2024 9,157 4 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

How to determine positive-stained cells in FACS? Use isotype or unstained control?

To compare positive and negative cell populations in flow cytometry, should I compare unstained cells with antibody stained cells? Or with the isotype control? Most papers show comparison with...

06 August 2024 6,728 6 View

Why do we equate male and female arousal?

Women, on the other hand, can become physically aroused (increased blood flow in the reproductive organs) without becoming psychologically aroused even in the slightest. (Robert Weiss)

05 August 2024 9,537 2 View

Low-yield gel extraction problem?

I am having an issue with my gel image where my PCR product is not appearing very bright on the gel. When I perform gel extraction, the A260/280 purity value is very low. I used the Qiagen gel...

05 August 2024 9,798 3 View

Is it true that the vacuum bomb is about to arrive?

We assume this to be true. We also assume that the vacuum bomb is the latest version of explosives with an explosive power of a few to ten kg of TNT equivalent. It has the unique characteristic of...

04 August 2024 4,534 1 View

How to make a Lateral flow control line more uniform?

We have a lateral flow test on Sartorius 140 NC. The conjugate is gold-monoclonal antibody. Using different control lines (GAM, protein A, protein G) we get a very strong leading edge with reduced...

31 July 2024 4,510 3 View

Suggestions for the Annexin V Positive Control ?

Hello everyone, I am currently using washed human platelets to stain Annexin V as a procoagulant marker. Additionally, I am staining with PerCP-CD61 to identify platelet cells. So far, I have...

29 July 2024 8,624 1 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