Is it possible to see downregulation of a gene (Ki67) on RT-qPCR while seeing an upregulation on immunofluorescence staining?

15 March 2024 0 1K Report

Hi everyone,

I performed an immunofluorescence (IF) staining for Ki67 using a validated, specific Ki67 antibody (Dako) and I see a clear upregulation of the protein expression compared to my baseline samples. However, on RT-qPCR, Ki67 is downregulated. Is this possible or should I question my staining (although the IF signal seems very specific to me)?

Thank you for your help!

Sara

Badges
Science method

More Sara Akalay's questions See All

Do I need to strip the PVDF membrane before staining it with colloidal gold?

Dear Colleagues, After running Western blot on PVDF membrane and detection using ECL, I would like to stain my PVDF with colloidal gold to be able to allign the ECL image with total proteins on...

02 March 2021 7,829 3 View

Shall I skip the measurement model in SmartPLS?

Hello everyone, I had run a mediation model in SmartPLS using questionnaires that were already validated among my statistical population. A professor who edited my paper -though she is not...

01 March 2021 3,052 3 View

Have anyone checked before Cyclic voltammograms of intial ITO substrate and E-teck?

As a beginner in electrochemistry, I tried to obtain cyclic voltamograms of commercial anatase powder deposited on an ITO substrate or on Eteck. After several attempts, I noticed that my cyclic...

23 February 2021 3,175 3 View

Is it okay to mention you distributed your qualitrics study link through social media sites?

Just want to be clear and know it is okay to say I have distributed the qualitrics link through social media such as Facebook, Instagram and Twitter to ensure my target audience is reached in my...

21 February 2021 5,143 2 View

Why samples do not run in resolving gel?

Hello everybody, I have a problem with SDS-PAGE and I do not know why my samples stop at the top of the resolving gel and they do not run through it. The percentage of gel is 15% and the size of...

14 February 2021 9,116 6 View

Do you know of an informal caregiver of a person who has recently received a bone marrow transplant?

We are conducting a pilot study of an intervention for informal caregivers of persons who have recently received a bone marrow transplant in the US. If you know of anyone who might be interested,...

13 February 2021 9,174 1 View

Is it possible that the single-pass absorption exceeds 50%?

Dear researchers It's well-known that the maximum light absorption of any structure can not be greater than 50%. Is it possible that the single-pass absorption exceeds 50%? How?

17 January 2021 2,335 7 View

Most appropriate fit of a standard curve in an enzym-linked lectin assay?

Dear all, I have a question regarding the most appropriate fit of a standard curve in an enzyme-linked lectin assay set-up to quantify the amount of mucin secretion in conjunctival explant...

03 January 2021 1,010 6 View

How do I access Domestic Violence Attitude Scales or find them for free?

Hi, I'm a postgraduate student currently starting up a research project on attitudes towards domestic violence (DV). The study will aim to compare cultural differences between attitudes and...

31 December 2020 781 3 View

A good protocol for protein purification by AKTA?

Hi everybody, Does anyone have a good protocol for protein purification using AKTA? I have tried to purify two proteins but after several attempts it was not successful. So, I really appreciate...

28 December 2020 5,220 15 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

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

Interested to stain mice tissue with WGA 488 conjugate, anybody has a protocol to recommend?

Interested to stain mice brain tissue with WGA 488 conjugate, appreciate if anyone willing to offer me some guidance or suggestions! Thank you!

28 February 2021 3,951 2 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

B-mercaptoethanol (BME) protocol?

Hi everybody, I have been running a multiplex IHC which allow me to stain the same tissue with different markers (Primary Ab + Secondary conjugated Ab) (AF487, AF555, AF647). Since I would like to...

27 February 2021 5,028 2 View

Why do my MCFs not form monolayer?

Hello, I am looking for some expertise on culturing MCF7s. I purchased them from ATCC and I have been following all media and culture conditions. Media is EMEM with 10% FBS and 10ug/mL of...

27 February 2021 3,904 2 View

Is there anyone who experienced with cytotoxic effect of Hoescht 33342 dye?

Is Hoechst toxic to cells? In general, it is known as non-toxic. I am working with dsRED MDA-MB-231 cell line. I applied it for my PDMS-based microfluidic platform, for this reason, I incubated...

26 February 2021 9,119 3 View