Jaya Aseervatham

22 Questions 61 Answers 0 Followers

Questions related from Jaya Aseervatham

Can you superimpose immunofluorescene images?

Hi All, I would like to know if there is a way to superimpose serial sections of tissues after doing immunofluorescence. The problem is I want to see the co-localization between few proteins and...

23 September 2020 1,214 3 View

Has anyone used R18 peptide for 14-3-3 inhibition in invitro?

hi, Is there anyone out there who has used R18 inhibitor peptide for inhibiting 14-3-3 in vitro. If yes could you let me know the concentration and does it need any peptide transfection reagent...

05 November 2019 3,630 0 View

Anone facing phototoxicity in IF samples?

Hi, Iam doing IF in HeLa cells and facing a weird problem. I use the standard protocol and when I expose the cell to laser light in the 488 channel, all the fluorescence fades the instant its...

30 September 2018 2,606 3 View

How to represent western blot for paper ?

Hi, I am trying to represent my western blots for paper and have a clarification how to represent the Y axis in the graph. I normalized the test protein with actin and then divided my test protein...

28 September 2017 7,934 4 View

Is there a Competetive peptide for GFP?

Hi, Iam using GFP nano trap from chromotek for my experiments and I want to study the structure of my protein by electron microscopy for which I need to get the protein in the pure form. After...

07 June 2017 1,616 2 View

Has anyone used GFP trap from Chromotek for IP?

Hi, Iam using the GFP Trap MA from chromotek for Ip of my protein. I followed everything as per the protocol, but was not successful. My protein was seen in the flowthrough, which means that it...

12 April 2017 4,696 21 View

What lysis buffer is recommended for membrane proteins for Co-IP?

Hi, I want to do a Co-ip of a connexin and its associated proteins for mass spec analysis. The problem is, to get the connexin and the bound proteins intact. Can anyone suggest a lysis buffer/...

07 January 2017 6,146 3 View

QPCR showing no difference in the knockdown and control?

HI, I am doing a lentiviral knockdown of a gene called DSPP and have prepared the stable clones which took me about 4 month with puromycin selection. The problem that I am facing is, when I do a...

24 June 2016 9,385 4 View

How to calculate the Multiplicity of infection (MOI)?

Hi, I have to do transduction using lentivirus and I have a doubt how to calculate the MOI. I have 5x10 3 ( 5000 viral particles/ ul). How much do I have to take to get MOI of 1 if I plate 10000...

28 January 2016 2,339 13 View

Problem with MCF 12 F breast cell line

HI, I am working on breast cancer and I\m trying to use MCF12F as normal breast cell line. I purchased it from ATCC and Iam having problem growing it. It dosent seem to attach and the few cells...

29 September 2015 3,218 7 View

What are the concepts of paraformaldehyde VS formaldehyde fixing?

Hi, I would like to know if any one has tried formaldehyde for fixing vs PFA. I would like to know the difference in the primary and secondary binding when both the reagents are used. Jaya

07 August 2015 927 11 View

Why does my secondary antibody conjugated with alexa 488 show fluorescence on cells?

Hi, As the follow up of my earlier question about the negative control showing fluorescence, Ihave found out that the secondary conjugated Alexa 488 is showing the fluorescence in the cells. I did...

29 July 2015 4,545 32 View

How is it possible to get fluorescence in IgG in cells?

Hi, Iam working with a cervical cancer cell line and Iam supposed to do immunofluorescence for MMP's. Iam facing a wierd problem. When I do the negative control with IgG, (Alexa 488) there is a...

23 July 2015 8,165 6 View

Can caspases activity decrease in apoptosis ?

Hi, Iam working on a oral cancer cell line and Iam facing a problem. The DNA damage shows apoptosis but the caspase 3 and 9 levels are low. Is this possible or what Iam seeing is an artifact....

11 July 2015 4,435 10 View

Can someone help with a problem on a stable clone ?

Hi, I am having a stable clone having G418 resistance and puromycin resistance. After expanding the cells, I freezed using 50% FBS, 40% medium and 10% DMSO, left the cells in -80 and then froze in...

08 March 2015 7,673 8 View

How can I find the interaction between two proteins?

Hi, I would like to know, if there is a way to find the interactions between two proteins. Iam working on a dental protein called Dentin Matrix protein and loss of this activity is reported to...

13 January 2015 9,457 3 View

Can I loose the protein of interest from a stable clone?

hi, Iam facing a weird problem after transfection. I am using C3H10 meschencymal cell line which is a stable clone expressing a protein which is GFP tagged. Iam using this cell line to transfect...

15 December 2014 3,946 15 View

Why am I not able to get rid of Bacterial contamination in cell culture?

Hi, I am having bacterial contamination in cell culture. I see these wriggling at high magnification and as black balls at low magnification. My problem is, I see these only after I start...

11 December 2014 5,308 6 View

How can one minimise more non specific bands on western?

Hi, I am using an antibody which is indigenous. The problem is it binds non specifically to more proteins. I see around 8-9 bands on the western. Even if I use more dilution this same pattern is...

02 November 2014 7,556 16 View

How to minimize cell death after transfection?

Hi, I am using MC3T3E1 cells ( mouse preosteoblasts) for transfection with Xtreme9 reagent. 24 hrs after transfection I see lot of cell death. Is there a way to minimise it? Thanks

19 September 2014 1,944 3 View

Does the absence of cyclin B1 in western mean that the concentrations used were too low?

Hi, I am using nocodozole (20-100ng) and colchicine (0.5-2.5uM) on C3H10 cell to arrest the cells in G2/M phase for 18-24 hrs. Then I do a western for cyclin B1. What I found was that there was no...

19 September 2014 3,211 3 View

Non specific binding of licor secondary antibody?

Hi, Iam using donkey antirabbit secondary antibody at the 700 channel to detect my protein of interest (1;1000 dilution primary ). My problem is that the green channel is showing non specific...

01 January 1970 3,251 2 View