Which buffer is best to achieve permeabilization for Ki67 FACS?

07 July 2014 1 10K Report

Hello everyone,

which buffer do you prefer to use for cell permeabilization in FACS: "PBS with 1% FBS, 0.09% NaN3" or "PBS with 0,25% Triton X-100?!

Anne von Koschembahr

I have not performed Ki67 staining but I have done BrdU staining by FACS. The permeabilization buffer for that is used is 2N HCl with 0.5% Triton X-100 for 30 minutes at RT. Following centrifugation, cells are neutralized in 0.1M sodium tetraborate with 0.5% Tween 20. We use this for primary human skin cells.

Badges
Science method

More Amir Mofidi's questions See All

For how long can we keep blood sample in an incubator?

Hi everyone, I am going to collect human blood sample in EDTA contained tubes and irradiate them. To analyze the impact of irradiation on plasma, I need to incubate the irradiated samples...

06 July 2018 314 0 View

Can anyone recommend a Phosphorilated Housekeeping gene for Western Blot?

I'm looking for an intra cellular protein phosphorylation through Western Blotting, as I'm following a protocol specific for phosphorylated protein, I can not use GAPDH and b-Actin as housekeeping...

11 December 2013 8,264 5 View

How to compare two groups with only two measurements?

Hiiiii everyone! I have an enquiry on statistical analysis. I was looking for many forum and it's still cannot solve my problem. I want to compare means of two groups of data but only with two...

03 March 2021 8,796 3 View

Why does blocking with my immunogenic peptide cause nuclear fluorescence in ICC?

Hi all, I am validating a polyclonal antibody raised against human ANT4, through WB and ICC. In most of my controls, the antibody shows very little off-target binding, but when I pre-incubate the...

03 March 2021 5,999 3 View

Enhanced Yellow Fluorescent Protein (Aequorea victoria) DNA sequence?

I am on the lookout for the Enhanced Yellow Fluorescent Protein (Aequorea victoria) DNA sequence. Does anyone know where I can find it? Thank you in advance

03 March 2021 3,568 1 View

What kind of fluid could I use for a pitfall trap that also does not invalidate molecular testing?

Hi, I want to start testing pitfall trap to obtain ants samples, but I need to conduct molecular analysis on those insects. So, what kind of fluid can I use? Ethanol expires too early and I need...

03 March 2021 5,978 5 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

What's the best way to measure growth rates in House sparrow chicks from day 2 to day 10?

What's the best way to measure growth rates in House sparrow chicks from day 2 to day 10? Since, the growth curve from day 2 to 10 won't be like the "Logistic curve" it might not follow logistic...

03 March 2021 1,401 3 View

After a reference from Int J Body Compos Res, can any body help?

Hi, I am after the reference below, my library says it cannot obtain a copy either locally or internationally, any help appreciated! Chris Wang ZM, Heshka S, Wielopolski L, Pi-Sunyer FX, Pierson...

03 March 2021 6,193 1 View

Can we block unwanted images during the x-ray prediction?

i am try to classify the x-ray images. During classification , can i block unwanted images (except x-ray image).

03 March 2021 7,100 1 View

Do we need to write protocol for a second study of a systematic review that has been registered and published?

I have conducted and published a systematic review and meta-analysis research with the topic related to public health and health pomotion (protocol was registed in PROSPERO). Now we want to...

03 March 2021 8,920 3 View