Filip Garbicz

13 Questions 50 Answers 0 Followers

Questions related from Filip Garbicz

Which WB STAT3 and STAT5 antibodies are best?

Hi, I'm currently having problems with Western blots using the following antibodies from Cell Signaling: Phospho-Stat3 (Tyr705) STAT3 (total) Phospho-Stat5 (Tyr694) STAT5 (total) They seem to...

08 August 2019 6,021 5 View

Do you know a good inducible triple-shRNA retroviral vector?

I'm looking for a reliable retroviral vector + that would allow me to express three different shRNAs simultaneously from the same plasmid + the shRNAs would be under the control of an inducible...

05 May 2018 3,366 2 View

How can I efficiently isolate CD138+ cells from myeloma patients' bone marrow samples?

First, I'm using Ficoll Histopaque centrifugation to obtain mononuclear cells. Then, I'm doing RBC lysis. I observe a drastic loss of CD138+ cells after that (as checked using FACS and two...

12 December 2017 6,649 13 View

Does anyone have suggestions on NGS Whole-exome sequencing - controversial thresholds?

A recent paper searching for novel mutations using whole-exome sequencing with preamplification assumed the following thresholds:> 6 (and 20> 10%...

04 April 2015 1,796 7 View

Circulating micro-RNA isolation - which method do you prefer?

I am new to the field of circulating nucleic acid detection. I would like to search for microRNA down/upregulation in blood samples from patients suffering from certain type of cancer. I have...

03 March 2015 9,906 9 View

Which reporter plasmid do you recommend for mutated gene transcription analysis?

I have found a mutation that probably decreases the level of transcription of my gene of interest. I would like to clone a part of the gene into a plasmid that would contain a reporter gene, so...

11 November 2014 3,331 9 View

Can anyone verify whether this is an artifact or an insertion in these Sanger sequencing results?

Not having much experience with sequencing, I would like somebody to look at the attachment. According to the sequence results, there is an insertion between 68G and 70A. That cytosine peak is not...

03 March 2014 3,588 26 View

How can you measure the activity of ONLY extrinsic or ONLY intrinsic pathways in apoptosis?

I would like to measure the activity of FAS/TNFR-dependent pathways and apoptosome-dependent pathways separately using FACS or a fluorometer. I do NOT want to use Western blot. The best solution...

03 March 2014 2,701 21 View

Has anyone experienced very low transfection efficiency using Lipofectamine 2000?

There is a major problem with transfections in our project. I have recently tested Lipofectamine 2000 and a Cyan Fluorescent Protein (CFP) plasmid on CHO adherent cells. According to the protocol,...

02 February 2014 8,320 28 View

H9C2 cardiomyocytes - Lipofectamine 2000 transfection conundrum?

I would like to know if any of you have ever worked with H9C2 cells and successfully transfected them. Alternatively, has anyone worked with any type of cardiomyocytes and performed a successful...

11 November 2013 8,933 8 View

Confocal microscopy novice - can anyone help with fixing and permeabilizing?

We're trying to observe BAG3 expression in H9C2 cardiomyocytes using a primary Alexa Fluor 488nm anti-myc-Tag Ab (the BAG3 is tagged with myc and is expressed from a transfected plasmid). We tried...

11 November 2013 9,844 15 View

What are the domain differences between Uniprot and NCBI?

Iam a student, just started working with protein mutations and I used NCBI info about different domains present in ITK. I used NCBI because it had much more data e.g. PI binding sites. However,...

08 August 2013 4,700 4 View

Has it been established, what is the effect of cholecystokinin on gastric motility? Does it cause contraction, or relaxation of its muscle?

I'm sure it inhibits the emptying of the stomach. However many sources claim that it is done by contraction of the muscles, and others say that its due to relaxation via vagovagal reflex.

05 May 2013 8,490 4 View