Immunology Techniques

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What is the lowest pH you would recommend using in IP injections--that will not cause pain to the animal?

I am injecting mice IP with the drug vinpocetine. Combing through the literature--it appears that most researchers use a vehicle of saline acidified with HCl to a pH between 4 and 5.5. These seem...

07 July 2015 7,894 3 View

How can I measure concentration of antiVirus IgA in mouse serum?

Hi, Is there any way to calculate actual concentration of antivirus IgA/ IgG/IgM in mouse serum using ELISA method? Can I use any indirect method for this, e.g. using mouse IgA purified protein as...

07 July 2015 1,901 2 View

What is the best marker for neutrophil identification by immunohistochemistry?

I want to identify neutrophil in rat lung tissue by immunohistochemistry. What is the best marker for identification of neutrophil in lung tissue by immunohistochemistry?

07 July 2015 2,770 11 View

Is it expected to get high perforin with normal degranulation on NK cell along with high cytotoxicity?

I am working with NK cell cytotoxicity. We found high perforin on NK cells along with cytotoxicity but a normal degranulation in antigen treated PBMCs compared with mock. Any suggestion regarding...

07 July 2015 6,652 2 View

Can anyone recommend a protocol for isolation of neutrophils from mouse liver?

Can anyone recommend a protocol to isolation neutrophils from mouse liver? Thanks Ilyas

07 July 2015 3,556 3 View

When culturing human monocyte derived macrophages, which is better: FBS or human serum?

I want a M0 macrophage population. Thanks!

07 July 2015 9,837 8 View

ICan anyone help with gG labelling with HRP protocol without colum seperation?

Dear all, I want to label IgG with HRP, the following protocol have to seperate labelled antibody with colum, which may cause sample lose. Does anyone have procols avoiding colum seperation steps...

07 July 2015 7,675 5 View

Can anyone recommend a protocol for Jurkat T-cells stimulation with soluble anti-CD3 and anti-CD28 antibodies?

I am looking for a protocol for the stimulation of Jurkat T cells with soluble anti-CD3 and ati-CD28 antibodies. I am working on the T-cells stimulation and effects of on kinectics. In my protocol...

07 July 2015 6,519 4 View

Is there any possibility for staining TLR4 only on cell membrane?

I need to do TLR4 staining on cell membrane. Every time I did it without any permeabilization, TLR4 in cytoplasm has also been stained. I have already tried PFA(4%) fixation with different...

07 July 2015 8,961 4 View

If you have an immunogenic Ookinete protein, how do you go about testing the functionality of its antibody and its viability in transmission blocking?

I will be happy if you could send me fultext articles with detailed methodology and discussion

07 July 2015 6,030 2 View

Is anyone working on macrophage processing of antigens resulting in antigen-specific immune RNA?

When such iRNA is incubated with B-cells they produce antibodies to such antigens they have not ever been exposed to.

07 July 2015 8,342 0 View

Is it possible to create a immuno-silent VLP?

I have been trying to find a way to produce a non-immunogenic variant of either HBsAg-S or HbC VLP carrier to express and demonstrate a protein of interest. I will use this VLP with antigen to...

07 July 2015 9,722 2 View

How can I peform IHC for CD3, CD4 and CD8 with frozen sections or PFA?

Dear Collegues, I have been trying to do an IHC in lungs with both frozen (non fixed before freeze) and tissues fixed in 4% PFA. I am not having positive results with any of these methods. Can...

07 July 2015 9,532 11 View

Has anyone used the PCR machines and gel documentation systems made by Labnet?

These equipments from Labnet look very enticing, but oddly, seem very unpopular, as none of my colleagues knows about them, and I could not find any customer reviews. Any feedback highly appreciated.

24 June 2015 5,523 3 View

Would endotoxin also stimulate irradiated PBMC?

I know that endotoxin (at levels 10-100 Eu/mL) in protein samples would not affect /stimulate purified T cells, but will stimulate PBMC. However, if people activate antigen-specific T cells with...

23 June 2015 8,927 3 View

What is the optimal enzyme concentration for immunoassay?

hi everyone, My assay setup is bare surface--->antibody---> biotin labelled analyte--->streptavidin labelled alkaline phosphatase---> substrate--->read signal I am using antibodies...

06 June 2015 1,991 2 View

Does anyone have a better method for generating BM derived pDCs than driving the culture with Flt3L? Or advice to get the cell yield above 10%?

My normal protocol is Flt3L (100ng/ml) supplemented culture media (10% FCS, PSG and 2me in RPMI) driven for 7 days (half the media replenished at day 3 or 4). Any reference to a better method or...

06 June 2015 8,644 2 View

What's the "normal" percentage range for PAN-T cells out of PBMCs?

Hi, colleagues. Unfortunately I have just started to work in this new field for me, PBMC purification from blood bags, MACS separation etc. So what I do is out of a blood bag from hospital I do a...

06 June 2015 5,918 8 View

How can I remove formalin from the final product in vaccine production ?

http://www.ncbi.nlm.nih.gov/pubmed/6873478 I am facing a problem in formaldehyde removal through diafiltration from whole cell vaccine.

06 June 2015 7,281 12 View

Would endotoxin also stimulate irradiated PBMC?

I know that endotoxin (at levels 10-100 Eu/mL) in protein samples would not affect /stimulate purified T cells, but will stimulate PBMC. However, if people activate antigen-specific T cells with...

06 June 2015 5,941 3 View

How can I quantify nuclei isolated from monocytes?

Hi all, We are trying to perform a 4C (Circularized Chromosome Conformation Capture) analysis on monocytes isolated from buffy coats. Just a basic overview of the approach, we perform a standard...

06 June 2015 2,447 4 View

Has anyone used the PCR machines and gel documentation systems made by Labnet?

These equipments from Labnet look very enticing, but oddly, seem very unpopular, as none of my colleagues knows about them, and I could not find any customer reviews. Any feedback highly appreciated.

06 June 2015 6,625 4 View

Why do I have so little events compared to the number of cells I plate?

I plate around 800,000 cells (PBMCs) in a well in a 96well plate, but when I read them on the LSR-II about 4 days later I only get 250,000-150,000 events total, half of which is viable, only 10%...

05 May 2015 6,264 13 View

What's the difference of anti human antibody and anti mouse antibody ?

I want to stain the human TIL with cell marker antibody.When I try to order the antibody..I notice that some of the antibodies named anti-human antibody and some named anti-mouse antibody..It's...

05 May 2015 617 5 View

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