Sudheer Babu

19 Questions 11 Answers 0 Followers

Questions related from Sudheer Babu

Dimer noticed instead of monomer on western blot?

Hi, i performed western blot to determine the protein expression. I expect a band at 22 kDa, interestingly the strong protein band noticed as dimer at 44 kDa. May i know what could be the...

last month 5,229 1 View

Top marker bands on pvdf slighlty moved during western blot transfer?

Hi, May i know why did i get the top marker bands slightly moved on pvdf memrbane while transfering the western blot? And do filter paper, gel and pvdf membrane should be similar size? if i run...

last month 6,795 1 View

Isolating proteins using trizol method?

Hi, I wanted to isolate proteins using trizol method. During performing the method, i was unable to dissolve the pellet in buffer comprising 0.3 M Gn-HCl, ethanol, 2.5% glycerol. However i...

07 August 2025 5,495 2 View

Why am I obtaining more or less than 100% if sum up all cell cycle phases with Flowjo?

Hi, I have a query relevant to flowjo cell cycle analysis. I obtained certain times 96% or 105% instead of 100% when i sum up G1, S and G2 phases, how could i solve the issue? or will be a problem...

24 July 2025 128 1 View

Calculating mean and SD from four biological replicates?

Hi I have a data from four biological replicates, each biological replicate contains 3 technical replicates. If i want to calculate mean and standard deviation from four biological replicates, do...

22 May 2025 2,927 3 View

Bonferroni correction for unpaired t-test for two independent groups?

Hi, I have a query regarding Bonferroni correction. I have three independent experiment groups 1) Control, 2) treatment-1, and 3) treatment-2 and performed unpaired t-test for control vs...

14 March 2025 6,604 3 View

Does the incubation at 37 degree needed for Propidium iodide/RNAase A solution in flow cytometry?

Hi, I wanted to analyze cell cycle for HaCaT by flow cytometry with propidium iodide staining. After fixation, do the cells incubate at 37 degree for 30 minutes initially with RNAase A followed...

06 February 2025 4,415 2 View

Loading control for western blot (image attached)?

Hi, Procedure to isolate total histones: 1) cell pellets were resuspended in 1 ml of hypotonic solution. 2) The nuclei were pelleted and sulfuric acid (0.2 M) was added to the pellet to extract...

04 February 2025 1,273 0 View

The bottom of the resolving gel of sds-page forme bubbles (image attached)?

Hi When i casted 14% resolving gel of sds-page, the bottom of the resolving gel formed bubbles with solidification (image attached), may i know what is the reason and how to solve the...

19 January 2025 3,431 1 View

Continuous western blot bands without space (image attached)?

Hello, I obtain western blot bands at desired region on the membrane but there is no space between the bands (attaching image for reference), may i know what could be the reason? TIA

15 October 2024 7,038 4 View

Storing harvested mammalian cell pellets?

Hi, I culture HACAT mammalian immortalized cells. I am interested to check the protein changes by western blot and cell cycle analysis by flow cytometry. Can i store the harvested cell pellets in...

13 August 2024 8,997 3 View

Loading control for western blot to detect Histone H1.2?

Hello, I isolated total histones successfully from the skin primary cells by acid extraction. I wanted to detect histone H1.2 expression in different conditions by western blot. Can some one...

11 July 2024 9,299 1 View

Silence epigenetic modifications

Hello, Is it possible to silence all kinds of epigenetic modifications in a cell by using gene or inhibitor? Thank you.

01 January 1970 9,779 2 View

Histone isolation via normal lysis or using acid, Hcl or Sulphuric acid?

Hello everyone, I wanted to detect histone protein expression levels in untreated and treated cell culture samples. Few research articles demonstrate histone can be isolated by usual lysis...

01 January 1970 2,496 1 View

How many cells need to be analysed from microscopy images to measure fluorescence intensity?

Hello, I performed immunocytochemistry and captured images with confocal microscopy. I wanted to measure the fluorescence intensity of different experimental conditions, may i know how many...

01 January 1970 8,884 2 View

Normalization of western blot with total protein Coomassie stained gel?

Hi, Shall i normalize the western blot protein bands with total protein gel stained with coomassie? Thank you,

01 January 1970 3,812 1 View

Genes involved in keratinocyte proliferation using bioinformatic tools or methods?

Hi, I am a layman to bioinformatics. I do not have an experimental data, curious to explore all the genes involved in keratinocyte proliferation or cell division in an image format employing...

01 January 1970 7,633 0 View

Fixation and storage of fixed cells for flow cytometry analysis?

Hi, To determine the cell cycle analysis by Flow cytometry, after trypsinization of the cells, fix it in 75% ethanol for 30 minutes on ice, subsequently store the fixed cells with ethanol at -20...

01 January 1970 307 0 View

Cells storage temperature after fixation with 70% Ethanol?

Hi I work with HACAT cells and wanted to analyse the DNA content of cells by DAPI with flow cytometry. After harvesting, the cells were fixed with 70% ethanol on ice for 30 min. After fixation,...

01 January 1970 5,557 1 View