What is the percentage expression of CR3 (cd11b/cd18) receptor on Macrophages?

I am checking CR3 expression on PMA stimulated THP-1 macrophages. In my experiment i used antibodies against CR3 and checked expression by flowsight. I got nearly 20% control THP-1 cells...

01 February 2018 6,188 0 View

What is the simple and easy protocol to isolate RBC's from whole blood sample?

I have four specific questions regarding RBC's isolation from blood? Procedure-Procedure to isolate RBC's from whole blood? Can i use ficol method to isolate RBC's? my real concern for this...

31 December 2017 2,185 1 View

What is the role of Phorbol 12-myristate 13-acetate (PMA) in the induction of Complement Receptor 3 (CR3) or CD11b/CD18 and CR4 or CD11c/CD18?

Role of PMA on the expression and function of CR3 and CR4 expressed on innate immune cells.

04 May 2017 7,216 2 View

I want to ask about statistics used in analyzing growth curves of the microorganism?

I have two strains A and B. I grow them in a media for 0-24 hrs and took reading at every 30 mints. I plotted these values on a graph (Y=O.D and X=Time). I want to see the significance difference...

01 February 2017 7,926 3 View

Why membrane associated complex (MAC, C5-9) of compliment system can't form pore in cell wall of Candida?

03 April 2016 4,109 2 View

Does anyone have a recommendation for yeast reporter gene plasmid/protocol?

Hi, could anyone recommend a plasmid and/or protocol for reporter gene assay in S. cerevisiae? I want to assess the effect of growth conditions on a transcription factor, so I want to clone it`s...

01 March 2021 210 1 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

Expression cloning by using cDNA,do I need to modify the cDNA first?

I am going to have a expression cloning of mammalian gene by using shuttle plasmid to transforming the E.coli However I don't know I should only inserting the Coding sequence ,or I can...

28 February 2021 5,440 3 View

Problem with shRNA transfection. Why are only two plates getting contaminated instead of all?

I transfected my LNCaP-WT cells with 3 shRNA plus their NTC two weeks ago and split two puromycin selected cell plates on Friday last week(Feb 26). I checked for GFP in the cells, and they all...

28 February 2021 4,949 3 View

Anyone that can help with details in phosphopeptides enrichment?

Anyone know the amount of protein before enzymatic digestion for phosphopeptides enrichment of serume sample in Ti-IMAC? Has anyone ever use Ti-IMAC(the material is from Chinese company named...

28 February 2021 7,310 3 View

PUC19 as a transfer plasmid for lentiviral delivery?

Hello, Is it possible to use pUC19 as a transfer vector to be packed in using the second generation viral particles packaging system( pMD2.G; psPAX2 plasmids)? As far as I understand it there is...

28 February 2021 4,868 2 View

Did anyone ever try using pLVX-TetOne Plasmid with a copGFP as a selection marker instead of Puromycin?

When using a lentiviral vector for inducible expression of genes using the Tet-On system. In the literature, I saw a widely used of pLVX-TetOne-Puro Plasmid, is there a reason why people prefer to...

25 February 2021 1,011 3 View

Why do the PCR bands from the cDNA templates (extraction from the cell after transfection) show different size from the band of the plasmid template?

After transfection with the plasmid ( linearized ) and subcloning of the cell lines, RNA was extracted from the cell and then reverse transcripted to cDNA. When PCR reactions were run to verify...

25 February 2021 5,712 3 View

How do I transfect a large plasmid into PC12 cells with nerve growth factor (NGF) receptor?

I have a large 16 kb plasmid, which I need to transfect into PC12 cells. Lipofectamin 2000 didn't work and with GFP alone the transfection rate is very low. I also tried the neon invitrogen...

25 February 2021 6,635 5 View

Problem with ligating 2.8 kb insert into 8.5 kb plasmid?

I am trying to clone 2 copies of the same mammalian gene into the pSF-CMV-Ub-Puro Ascl (contains HindIII, KpnI and NheI cut sites) plasmid. This is what the final product is supposed to look like...

24 February 2021 6,310 3 View