免疫学技术

MaterialsDMEM high glucose (Life Technologies Inc. #10313-021 or equivalent)Fetal Bovine Serum (Life Technologies Inc. #16000-044 or equivalent)L-glutamine (Life Technologies Inc. #25030-149 or equ ...

Reagents (StemCell Technologies Inc. # 03800) Medium A - Pre-fusion Medium and Hybridoma Expansion Medium (StemCell Technologies Inc. - # 03801)Medium B - Fusion Medium (StemCell Technolog ...

Mouse ImmunizationOrder 6 six week old Balb/C mice and let the ARC know they are coming. Have your antigen ready for when they arrive. Once they get there earmark the mice and perform a pre-bleed on t ...

If titre is positive do one of the following 2 weeks later:Boost tail vein with 20 ug- 50 ug of Antigen in PBS and proceed with fusion on day 4. OR Boost subcutaneously with 50 ug - 100 ug of Antigen ...

NOTE: The sooner after cell fusion you can do a limiting dilution the better your chances of retaining strong positive hybridomas. Remove spleen cells from 1 or 2 mice wash as for cell fusion (above) ...

Immunolabeling of thin sections for EMThin sections of biological material mounted on specimen grids can be easily labeled by floating them section-side down on small drops of antibody. This method wi ...

Materials 0.1M NaHC03 pH9 DMSO NHS-Biotin (N-Hydroxysuccinimidobiotin Sigma #H-1759) PBS Procedure Dialyze the sample against carbonate buffer. After dialysis adjust sample to 1 mg/ml (or less). Prepa ...

Protocol:All steps up to the dialysis at rt.Pour column in TBS (=0.15M NaCl 20mM TrisCl pH 7.4). We use a 5 ml column for 25 mls serum. Wash extensively in TBS after prewashing as indicated in the pro ...

You can test whether or not you have gotten an immune response to the peptide and how strong that immune response is by doing ELISAs against peptide conjugated to BSA. By conjugating to BSA you will e ...

Immunization of Mice:Injections should be made at intervals of at least two weeks.. We have been using either of the two following adjuvants with success: Freund adjuvant and MPL+TDM adjuvant ( RIBI I ...

These are basically Tim''s procedures and have been used successfully by numerous members of our lab and by others around us. A. Checking Peptide Thiol GroupsDo this before thiol coupling either to KL ...

Required solutions are given below.1. Cells should be plated in 35 mm dishes containing coverslips. Fix cells when the cell density is not too high so the cells will not be crammed together. Usually p ...

Procedure:1) Immerse 18 mm2 glass coverslips in EtOH. In the tissue culture hood individually pull out and carefully flame to sterilize. Allow to cool then place in 35 mm dishes or in 6 well plates ...

General remarks:Ultrathin methacrylate-(e.g.Lowicryl HM20 K4M Monostep polar and nonpolar or LR-White) or epoxy (Epon/Araldite Spurr) sections are transferred with a loop on poly-L-lysine coated round ...

DescriptionThis procedure describes how to process cells for immunofluoresence microscopy. Procedure1. Sterilize coverslips under UV light for 20-30ma. To increase cell adherence treat coverslips with ...

Antibodies to small peptides have become an essential tool in life science research with applications including gene product detection and identification protein processing studies diagnostic tests pr ...

In order to decrease the amount of nonspecific staining it is often necessary to preabsorb primary and secondary antibodies to yeast cells lacking the antigen prior to use. A 1:1 mixture of fixed yeas ...

The multiple copies of the HA epitope present in the HAT tag can be detected by the mouse monoclonal antibodies 12CA5 (Boehringer) and 16B12 (MMS101R; BAbCO Richmond California) (see Table 1). On West ...

This is the protocol to use with the Botstein anti-actin rabbit antibodies.Grow cells at the appropriate temperature to 5x10E6 in 5 mls YPD. Add 0.5mls 37 formaldehyde (best grade) and incubate on the ...

Grow cells at the appropriate temperature to 5x10E6 in 5 mls YPD. Add 0.5mls 37 formaldehyde (best grade) and incubate on the roller at same temp. for 10 min. Spin down cells 2Kx3min. and resuspend in ...