Herzenberg Procedures (From Binder)

Front

            B Cell Blind Panel Transfectant Results

            Old Table of Contents

            New Table of Contents (Word)

 

Serum Preparation

            Bleeding Procedures

            Immunization Protocols

            Allotype Immunization Results

            Production of Rabbit Antisera

            C' Absorption

 

Cell Preparation

            LeucoPrep from BD

            AO/EB

            FDA

            CRBC

            Bone Marrow Removal Technique

            Dissection of Mouse Spleen

            Cell Treatment With Glass Wool

            Preparation of Buffy Coat

            Counting Cells

            Counting Cells With an Electronic Counter

            Loading Cells onto Gradient

            Separation of Lymphocytes Using Ficoll-Hypaque

            Ficoll-Isopaque Technique

            Method for Velocity Sedimentation Ig Gradient

            Ficoll-Isopaque Technique (article)

            Identification of Dead Cells in FACS Cloning Procedure

 

Solutions

            Cell Counting Solutions

            Trypsinization Reagent

            Sodium Azide NaN3

            Hepes Buffer

            Balanced Salt Solution (BSS)

            ulbecco Phosphate-Bufferred Saline (PBS)

            Ethylenediamine Tetraacetic Acid (EDTA or Versene) pH 7.2

            Acid Citrate Dextrose Solution

            Procedure for IsoMEM Preparation

            Antibiotics for Irradiated Mice

            Preparation of Glutamine for Tissue Culture Media

            Preparation of Concentrated Pen-Strep for Frozen Aliquots

            To Calculate Molarity of a Solution When Only Limited Information is Given on Label

 

Separation of T and B Cells

            Procedure for Cell Separation by Nylon Wool Columns

            Dupont Procedure for Wool Columns

            Anti-Theta Treatment of Cells

            Hepes Buffer

            Preparations for Cell Separators

            Human White Cell Separation

 

Antigen Preparation

            TNP Conjugation to BRBC for Hemaglutination and Plaquing (updated for BRBC)

            TNP Conjugation to SRBC for Hemaglutination and Plaquing

            Preparation of KLH

            CoCl3 Coupling of HGG, KLH and POL to SRBC (Conjugate with CRCL3 on Day of Use)

            Preparation of KLH, HGG Coated Gluteraldehyde Fixed SRBC for Hemagglutination

            Preparation of DNP-KLH, HGG

            Alum Precipitation

            Use of Biotin-Activated Esters for Protein Conjugation

 

Culture of Lymph and Spleen

            Preparation of Human Leukocyte Cultures for Chromosome Analysis

            Mixed Lymphocyte Reaction

            Mitogen Simulation

            Preparation of Tissue Culture cells -H3 Thymidine-Pulsed Prior to Counting in Scintillation Counter

 

Ab Purification

            Ultrogel A - Ultrogel AcA Instruction Sheet

            Absorption of Antibody with Lymphoid Cells

            DE-52 Column Chromatography

            Sepharose 4B Conjugation

            Buffer

            Ion Agar Electrophoresis

            Electrophoresis

            Radio-immune Assay

 

Ab Assays

            Radioimmunoassay

            Quantitation of Immunoglobulins by Inhibition of Precipitation of I-125 Labelled Antigens

            Cunningham Plaque Assay

            Cytotoxicity Assay - Cr51 Release

            Hemagglutination Procedure

            Agar Plates for Phage Inhibition Assay

            Procedure for Phage Inhibition

            Media for Phage Inhibition Assay

            Phosphate-Gelatin Buffer (0.05M)

            Hemagglutination Technique

            Precoating and Pouring Agar Slides

            Ouchterlony Procedure

            Procedure for Mouse Ouchterlony

            Sample Ouchterlony Sheets

            Sodium Bicarbonate Wash

            Elispot Methods

            Mishell-Dutton Culture Methods

            IgG Elispot

 

Cell Staining

            Microtiter Staining Method

            Membrane Fluorescent Antibody Staining

            Mithramycin Staining Protocol

            Cytoplasmic Fluorescent Antibody Staining

            F-KLH Staining Procedure

            FDA Vital Cell Stain

            Stripping of Surface Globulin in Mouse Cells

            Pronase Stripping of Membrane Immunoglobulin in Rabbits

            ICN Biomedicals

            Wright Giemsa Stain (Blood Smears)

            Wright Giemsa Stain

            Autoradiography of Cell Smears

            Chemical Identification of Monocytes

            Cytochemical Identification of Monocytes and Granulocytes (article)

 

Labelling

            FTC Labelling in Suspension

            Fluoresceination

            Fluoresceinating C----valin A

 

Fixation of Cells

            Formalin Fixation of Cells

            Fixation of Cells with Glutaraldehyde

 

Radiolabelling

            Immunoprecipitation of Radiolabelled Proteins from NP-40 Extracts

            S35-Methionine Labelling of Lymphocytes and NP-40 Extraction of Membrane Proteins

            TCA Precipitation for Determinaion of Radioactivity

            Labelling with I-125

            Preparation of Iodogen Tubes

            I-125 Labelling of Proteins

            Solid Phase I-125 Labelling Protocol Page

 

Membranes and Microspheres

            Coupling Protein to Microspheres

            Binding of Microspheres to Hybrind Cells

            Dilute Procedure for Coupling

            Standard protein-Microsphere Coupling Procedure

            Isolation of Plasma Membranes

 

Miscellaneous

            Sepracell-MN Separation Manual

            Preparation of Brushite and Hydroxylapatite

            Mycoplasma Screening of Cell Lines Using H33258 Staining of a 3T3 Indicator Cell Layer

            Two-Dimensional Gel Electrophoresis

            Operation of Packard Liquid Scintillation Counter

            X-Irradiation Procedure for Mice

            Preparation of Polaroid 3x5 Lantern Slides

            Use of Cytocentrifuge

            Operation of Cavalli's Cell Freezing Apparatus

            Use of Osometer

            (Microscope) Photographing Settings For Slides

            Changing UV Light on Wild Scope (Left Hand Scope)

            Changing Bulb on Right Hand Scope

            Washing Dialysis Membrane from Marbrook Cultures

            Physiochemical data of _____ used in the adoptive transfer of the secondary _____ immune response

            Myeloma Proteins = Groups = Allotypes (Class)

            Sterile Hood Inspection

            International Centrifuges - Nomograph for Computing Relative Centrifugal Force

            International Centrifuges - Weight per Microcurie, Half-Life Decay Chart, Counting Error, Conversions

            Instructions for H-P Calculation of I125 Assay

            Assay Method

            Mouse Skin Grafting Procedure (Aplit Thickness Method)

            Skin Grafting (New Procedure - 1981)

            Stock Room - Mating and Weaning

            Colormetric (MTT) Assay for Cell Survival and Proliferation

            Reagents for 2-D Gels

            Autoradiography of Cell Smears

            Preparation of Fluorescein or Rhodamine Labelled Chicken Red Blood Cells (CRBC)

            Isolation of Plasma Membranes

            Instructions for Growth of Rat x Mouse Hybridomas

            DNA Preparation