抗体抗原实验

Materials L-lysine coated glass cover slips or charged glass slides Neutral Buffered Formalin (Sigma HT50-128) 0.5% NP40 in PBS (2.5 N HCl or 0.07 N NaOH for BrDU staining only) Primary and secondary ...

Reagents and MaterialsPBSPBS: 0.05M EDTAPBS: 0.01 thimerosalConjugate Storage BufferSATADMFHydroxylamine hydrochlorideNaOH PelletsPD10 Column or Sephadex G25Sulfo-SMCCPreparation of Thiolated Ant ...

DescriptionThe Following Protocol can be used to couple either FITC or Biotin to IgG. The same method is used for both and when discrepancies occur they will be noted. Procedure1. Dissolve 2mg of IgG ...

DescriptionThe Following Protocol can be used to couple either FITC or Biotin to IgG. The same method is used for both and when discrepancies occur they will be noted. Procedure1. Dissolve 2mg of IgG ...

Solutions:-IMDM-IMDM complete + 15 MCM.-PBS-PEG 50 ( w/v ): PEG 4000 ( 50 ) from Boehringer Manheim 1 243 268-Aminopterin: Sigma A 5159 (50X )Prepare a T-150 flask with 170ml of IMDM complete with MC ...

ReagentsHorseradish peroxidaseSodium periodate 0.1M freshly madeEthylene glycolSephadex G-25Sodium acetate buffer 0.001M pH 4.2IgGCarbonate:Bicarbonate Buffer 1.0 M pH 9.5Sodium tetraborate (4 mg/ml) ...

Healthy Sp2/0 cells should be rapidly growing by this time. Sp 2/0 cells should be started about two weeks before the cell fusion. Every two days they should be centrifuged at a 64.4 xg on the IEC cli ...

Materials: P3X63Ag8.653 murine myeloma or YB2/0 (maintained at 50 w/v PEG 1500 warmed to 37° CMedium: IMDM supplemented with 20 fetal bovine serum 4 mM L-glutamine 1 mM sodium p ...

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 ...