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Questions related to Immunology Techniques
I'm trying to develop a chemi-luminescence based immunoassay for HIV antibody detection. For the purpose I'm using negative as well as positive control as a quality control. Currently, I'm using...
31 December 2019 2,694 3 View
I am looking for the activation of Jurkat T-cell and if you activate it then how can you measure their activation I could not find anything related to these following: CD69 IL2 The other...
11 May 2018 6,676 5 View
I am trying to find a kit to measure/detect endotoxin in purified protein samples. Here is a link to this vendor's...
05 April 2018 6,570 1 View
I am trying to culture the granulosa cell from mouse. When I use the JC-1 to detect its' mitochondrial membrane potential, I can see two different populations, one has low MMP, while another has...
24 July 2017 9,063 2 View
Hi all, At the the moment, I do co-culture experiments between T-cells and antigen presenting cells in 96-well U-bottom plate wells. These are good for co-culturing about 5x10^4 APCs per well....
11 July 2017 2,857 1 View
I am doing a time-course study on neutrophil apoptosis in cell cultures . So far we have been able to isolate the neutrophils from peripheral human blood with a high yield and viability . However...
26 June 2017 1,537 2 View
Which percentage of labeled T cells should I find in the peripheral lymph nodes after adoptive transfer of say 5-10 million labeled T cells? Is there a difference in efficiency between the...
16 June 2017 642 1 View
Hi, I need a protocol to this aim, many years ago I saw a very easy protocol to expand lymphocytes from PBMC using a Flu peptide, but now I have not access to this protocol. If someone has...
12 June 2017 8,820 4 View
I'm trying to make a Treg supression assay, to evaluate de ability of autologous Treg to inhibit the proliferation of Teff after treatment of patients with our product. To differentiate autologous...
05 April 2017 8,375 4 View
I'll dosage LPS with LAL QCL-1000 Chromogenic endpoint Lonza, and i have many questions about this. Has anyone used this kit yet? Does anyone know if all materials should be depyrogenised? Can the...
04 April 2017 7,829 1 View
I am attempting to stimulate mouse T-cell proliferation in vitro using plate-bound CD3 (10ug/ml, clone 145-2C11) and soluble CD28 (2ug/ml, clone 37.51) antibodies. I can induce some activation as...
04 April 2017 8,711 2 View
I;m working with THP-1 cell line. my problem is that from last ffew weaks cells are getting adhered to well plate after PMA stimulation (50 ng/ml) but didn;t get spread ir they are not showing any...
09 March 2017 8,401 3 View
I use RAW cells in DMEM + 10%FCS and "feed" them with heat treated (20 min 48C°) clatrin stained erythrocytes. I stimulate the RAW cells with 50ng/ml LPS and incubate the RAW cells and erys for...
09 March 2017 315 1 View
I derived M1 macrophage from human PBMC.Than I coated my CFSE stained thp1 target cells with my Abs on ice for 30mins, continue with 2 times washing in pbs----resuspend cells in IMDM (no...
24 February 2017 8,934 1 View
I am doing a validation test of an I-ELISA for B. canis and I experimented some problems with the ABTS, the negative controls are getting to high ODs. Have you experienced problems like this? How...
18 February 2017 2,470 1 View
I have no experience in this field and I really need to know if the results of my back titration are between 30 and 300 TCID50. How can I calculate the TCID50 that I obtained as a result of the...
30 January 2017 2,988 2 View
I want to perform an infectivity assay to test a metal compound and I need to guarantee that the medium hasn't it in a free form (apart from proteins). There is any protocol using a defined media...
20 January 2017 3,063 3 View
HI, all can we able to differentiate 5kb base pair difference with Blue native Page gel electrophoresis. Anyone has idea for some otehr technique to differentiate homodimer formation for NFkb1(100...
19 January 2017 2,821 1 View
Hello,I would like to do immunostainings on differentiated 3T3-L1 cells (adipocytes). We tried most of the "classic" protocols, fixation with either 4% PFA or cold methanol/acetone, and a lot of...
18 January 2017 8,648 3 View
Historically I have always used formaldehyde fixation of cells for immunofluorescence, but lately I have had to use methanol. All of the methods I have found state to do this at -20C, but none of...
18 January 2017 6,839 3 View
Simple question - I'm looking for recommendations for antibodies for RFP/mCherry immunoprecipitation. Thus far we've tried allele RFP nanobodies and not had much success. Do you have a go-to...
16 January 2017 5,994 3 View
I added 0.5 microgram LPS to DMEM+FCS+PS and then incubated 24h. After that, I transferred the LPS-stimulated DMEM to RAW264.7 macrophages and saw a stimulation in cytokine production. However,...
16 January 2017 7,206 3 View
Does anybody know a better way to separate areas in a slide for immunofluorescence assays than the Super Pap Pen by Life Technologies? Because this pen is quite dirty, easy to wash away and not as...
16 January 2017 7,997 5 View
I had practiced mouse hepatocyte isolation with percoll for several month. The primary hepatocyte cells can only survive one day or less, and the cause of cell death is DNA released....
16 January 2017 315 0 View