免疫学技术

Protocol for Limiting Dilutions 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 ...

Conjugation of Antibody to HRPAuthor: Nanci DonackiSource: Contributed by Nanci DonackiReagents and MaterialsPBSPBS: 0.05M EDTAPBS: 0.01% thimerosalConjugate Storage BufferSATADMFHydroxylamine hydr ...

Peroxidase Conjugation by Periodate MethodAuthor: Nanci DonackiSource: Contributed by Nanci DonackiReagentsHorseradish peroxidaseSodium periodate 0.1M freshly madeEthylene glycolSephadex G-25Sodium ac ...

Antibody LabelingImmunolabeling 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 antibod ...

Biotinylation of AntibodyAuthor: Nanci DonackiSource: Contributed by Nanci DonackiMaterials 0.1M NaHC03 pH9 DMSO NHS-Biotin (N-Hydroxysuccinimidobiotin Sigma #H-1759) PBS Procedure Dialyze the sample ...

The production of monoclonal antibody by hybridoma fusion: Immortalization of sensitized B lymphocytes from immune mice. OverviewWe outline a simple and contemporary protocol for the development o ...

Monoclonal Antibody ProductionMouse 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 mic ...

Fusion is an important procedure to produce monoclonal antibodies (MoAb).PROTOCOLprepare the culture of myeloma cells and maintain the concentration of the cells at the level of less than 10^6 cells/m ...

Recipies for Cell FusionPhosphate-buffered saline (PBS) (0.01 M Na2HPO4 0.15 M NaCl pH 7.4) sodium phosphate dibasic (Na2HPO4)0.87 g sodium phosphate monobasic (NaH2PO4 x H2O)0.54 gsodium chloride8.5 ...

General Fusion ProtocolMaterials: P3X63Ag8.653 murine myeloma or YB2/0 (maintained at /ml)50% w/v PEG 1500 warmed to 37° CMedium: IMDM supplemented with 20% fetal bovine serum 4 ...

Fusion of Mouse Rat or Hamster CellAuthor: Nanci DonackiSource: Contributed by Nanci DonackiMaterialsDMEM high glucose (Life Technologies Inc. #10313-021 or equivalent)Fetal Bovine Serum (Life Techn ...

GTPγS-induced Inhibition of Binding1. Chemicals and EquipmentDB Buffer PB cAMP cAMP 8- 3H (25 Ci/mmol 1.0 mCi/ml 32 000 nM) 10 mM GTPγS Wacker silicone AR-20 Reagiergefab test tubes (Sarstedt) millip ...

Enzyme-linked Immunosorbent Assays (ELISAs) combine the specificity of antibodies with the sensitivity of simple enzyme assays by using antibodies or antigens coupled to an easily-assayed enzyme that ...

Immunization Protocol for Monoclonal Antibody ProductionAntigens:Proteins: 10-100 µgCells: 0.5 - 5.0 x107Adjuvant:Complete Freunds AdjuvantNoneSchedule:DayManipulatio ...

GST-fusion protein injection into rabbitsConcentration of GST-fusion protein eluted from GSH affinity column The GST-fusion protein eluted from the GSH-sepharose is diluted and need to be concentrated ...

You'll need to couple the synthetic peptide hapten to a protein carrier with a bifunctional reagent. For antibody development we don't bother to use highly purified peptide. Crude (unpurified) peptide ...

Purification Of Anti Peptide AntibodyClaire WalczakJanuary 1996Protocol: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. W ...

Antigen Design & Sera Purification Tech Sheet Antibodies to small peptides have become an essential tool in life science research with applications including gene product detection and identification ...

If your peptide is synthesized with a terminal cysteine residue (i.e. a free sulfhydryl group -SH) you can use any maleimide-activated carrier protein to create an immunogen. One-step chemical reactio ...

Preparation Of Peptide-KLH Conjugates For ImmunizationClaire WalczakJanuary 1996These are basically Tim's procedures and have been used successfully by numerous members of our lab and by others around ...