抗原设计

ReagentsSodium Borohydride (NaBH4) - 1 mg/mlCarbonate:Bicarbonate Buffer pH 9.6 Sodium Periodate (NaIO4) - 1 mM 2.14 mg/10 ml Acetate Buffer 10mM Acetate Buffer pH 4.50.82 g sodium acetate in 90 ml di ...

Immunocytochemistry is the name given to methods that use antibodies to detect the location of proteins within cells. The antibodies bind specifically to the protein being investigated. With electron ...

This method has been used successfully in our laboratory since 1994. The protocol assumes all users will be familiar with routine TEM preparation methods. An important application of this method was m ...

Preparation of Colloidal Gold Conjugates.Colloidal gold has been used for centuries in the preparation of stained glass for windows and fine glassware. In recent years colloidal gold particles have be ...

Immunocytochemistry at the ultrastructural level (TEM)Results obtained from immunocytochemical labeling can often differ from what was expected. Sometimes this may result in the formulation of new the ...

Procedure Dilute p27 monoclonal antibody 1:1000 (v/v) in Carbonate Coating Buffer. Add 100 µl/well and incubate o/n @ 4°C or 1 hr @37°C. Wash 3X with TBST (pour onto plate empty into sink hit onto tow ...

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 ...