Why cannot i find my protein on cell surface after antibiotic selection of expressing plasmid?

More Subham Basak's questions See All

How can I generate random samples from Bingham distribution in R? How can I construct those symmetric matrices?

in the function rbingham(n,A) in R software how can I construct that A (symmetric) matrix?

24 February 2021 3,528 3 View

How to formulate Energy minimization in Molecular dynamics simulation for alloys using meam potential?

during MD simulation, the atoms are not moving as per required. The problem (as per my observation) lies in energy minimization technique. upon application of tensile strain in x or y direction,...

22 February 2021 380 5 View

How to generate AA6061 alloy or any aluminium alloy crystal in material studio?

I need to carry out molecular dynamics simulation on Aluminium alloy. For this i need the crystal structure that which could be further replicated to form a structure.

22 February 2021 1,410 1 View

How to prepare an algal sample for FT-IR spectroscopy?

Can we measure the bulk composition of biomolecules present within an algal sample directly by placing its powdered form on a FT-IR spectroscopy stage? Or does it require a particular kind of...

30 January 2021 2,303 4 View

what are the possible reasons for Crop Water Stress Index values higher than 1 under moisture stress condition?

I have determined CWSI using the theoretical approach of Jackson et al., (1982) based on canopy temperature measurement in chickpea. at the final stages of crop growth some of my values of CWSI...

28 October 2020 9,992 3 View

How can i increase biomass of my E. coli culture to better plasmid yield?

Hi everyone, I want to increase the biomass of my E. coli culture for better plasmid yield. Use of different strains of cells and media or any other suggestions are welcome. Thanks!

11 October 2020 6,400 3 View

Do you have the following alloys?

Dear colleagues, It will be a great help if someone can provide me small pieces (1.5 cm * 1.5 cm * 1.5 cm) of cast Al 5024 alloy and Al 7068 alloy (with and/or without T6 heat treatment. Happy to...

31 July 2020 3,593 1 View

What to do when there is no main or interaction effect but multivariate tests and pairwise comparision tables show significance?

I run repeated measures ANOVA for an intervention study. I have 3 intervention groups and 3-time points. The output in SPSS showed that there is no significant main effect of time and no...

02 April 2020 2,328 7 View

What is the reason behind using a Pt-Ir alloy as a tip material for STM (Scanning Tunneling Microscope) ?

For scanning tunneling microscopy it is generally proposed that that the apex of the tip consist of single atom. For a Pt-Ir tip prepared by mechanical cut the apex atom sometime be of Pt or that...

11 March 2020 4,746 3 View

Nonparametric equivalent of ANCOVA?

For testing the effectiveness of group intervention, I would like to conduct ANCOVA. I have one experimental and two comparison interventions. However, my data is not normally distributed. Also, I...

20 November 2019 432 9 View

What is the characteristic length to determine the Reynolds number for the scattered turbulent fluid flow?

Dear all, I just saw this example in FB on the difference between laminar flow and turbulent flow. The flow from a pipe hits the sink and scattered. The flow from the pipe is characterized as...

02 March 2021 1,999 5 View

Issues with Permeabilization during Flow Cytometry?

Hello Everyone. Currently I am working to characterize macrophages in the myocardium after ischemia-reperfusion injury in rats. Due to the low total cell number isolated from rat hearts I can...

01 March 2021 3,867 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

Precipitate after adding laemmli buffer to protein solution containing Potassium Acetate?

Hi, I am running a size exclusion chromatography experiment with a buffer containing Potassium Acetate as a salt. I analyse these fractions through SDS-PAGE. After boiling my SEC fractions in...

01 March 2021 2,622 3 View

Can someone suggest a way to prepare slides to observe under confocal microscope for observing fluorescent tagged proteins??

I am growing the cells on coverslips and transfecting the fluorescent tagged protein directly on them. Then i want to observe these cells under confocal microscope. Currently i am just using...

01 March 2021 6,142 3 View

What is the most suitable server for predicting protein structures in bacteria?

I have used the i-Tasser several times, however it has been unavailable for several days. I tried the swiss-model, but the output was not very pleasant due to the model used. Is there any other...

28 February 2021 4,521 3 View

Do you need the CMV enhancer before the CMV promotor for a proper function of CMV on the gene that follows?

I'm a student and I have to produce a cell line with a knockin for the NRF2 gene with GFP. I have to put a promotor in front of the GFP because the gene will be too far away from the promotor of...

28 February 2021 7,127 2 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

Problem with shRNA transfection. Why are only two plates getting contaminated instead of all?

I transfected my LNCaP-WT cells with 3 shRNA plus their NTC two weeks ago and split two puromycin selected cell plates on Friday last week(Feb 26). I checked for GFP in the cells, and they all...

28 February 2021 4,949 3 View

Any specific pipeline or parameter to analyse total transcriptomics study of brain tissue samples?

I have two groups of brain samples, control and treated for example. It was total RNA nova seq sequencing. I tried all the available pipeline like: star+rsem+deseq2, Hista+stringtie+cuffdiff,...

27 February 2021 356 6 View

Louis-Charles Béland

How was the expression of BCMA on your positive control in the FC experiment? Also could you confirm BCMA expression using another method (IF, WB) before FC?

Lenin Kumar Bompalli

please verify the cell line experimental condition, protein tagging and foldings

Subham Basak

Hi Louis-Charles Béland

I do not have a positive control cell line which i could use in the FC experiment.

I haven't done WB either, but I am going for WB confirmation now.