抗体抗原实验

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 ...

Testing Rabbit Bleeds By ElisaClaire WalczakJanuary 1996You 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 pep ...

This protocol describes a method for studying chemotaxis (migration towards a concentration gradient of chemoattractant) of leukocytes (neutrophils monocytes and lymphocytes) or other migratory cell ...

1. Resuspend cells (vegetative or developed) in phosphate buffer at 1 x 10 8 /ml .2. Add 50 ul cells to 50 ul 20 nM 3H-cAMP in 5-10 mM DTT (for background binding add same amount of hot cAMP plus 10 ...

CHIFF preparationDevelop cells for at least 3 hrs to minimize protease activity. Cell are resuspended in DB to 2-4 x 107/ml and shaken at 120 rpm at 22o for 3-6 hrs. Cells were harvested and washed wi ...

Sandwich ELISAThe sandwich ELISA measures the amount of antigen between two layers of antibodies. The antigens to be measured must contain at least two antigenic sites capable of binding to antibody s ...

MaterialsNUNC-Immuno Plate IIF. Horseradish Peroxidase-Streptavidin (Zymed; 43-4323) PBS or Balanced Salt Solution (BSS). Do not use RPMI or any medium that contains biotin. BSA (1% in PBS). ABTS subs ...

ELISA (Enzyme-Linked ImmunoadSorbent Assay)You will need the following solutions: PBS (Phosphate-Buffered Saline Ca/Mg free) :- Solute1x10xNaCl8g80gKCl0.2g2gNa2PO4 (anhydrous)1.15g11.5gKH2PO4 (anhydro ...

Gangliosides ELISA protocolAuthor: pingsunjimSource: Contributed by PingsunjimAbstract: This protocol can be used for detection of gangliosides.Spicific antibodies to gangliosides are added to cells a ...

Indirect ELISAThe indirect ELISA is used primarily to determine the strength and/or amount of antibody response in a sample whether it is from the serum of an immunized animal or the cell supernatant ...