Stanford
Herzenberg Laboratory

Protocols & Procedures

Here is the protocols & procedures information we have accumulated over the years.

Ab Assays

Agar Plates for Phage Inhibition Assay

Antibody Titration Curve

Cunningham Plaque Assay

Cytotoxicity Assay - Cr51 Release

Elispot IgG

Elispot Methods

Hemagglutination Procedure

Hemagglutination Technique

Media for Phage Inhibition Assay

Mishell-Dutton Culture Methods

Ouchterlony Mouse Procedure

Ouchterlony Procedure

Ouchterlony Sample Sheets

Phosphate-Gelatin Buffer (0.05M)

Precoating and Pouring Agar Slides

Procedure for Phage Inhibition

Quantitation of Immunoglobulins by Inhibition or Precipitation of I-125 Labeled Antigens

Radioimmunoassay

Sodium Bicarbonate Wash

Ab Purification

Absorption of Antibody with Lymphoid Cells

Buffer

DE-52 Column Chromatography

Electrophoresis

Ion Agar Electrophoresis

Radio-Immune Assay

Sepharose 4B Conjugation

Ultrogel A - Ultrogel AcA Instruction Sheet

Antigen Preparation

Alum Precipitation

CoC13 Coupling of HGG, KLH and POL to SRBC (Conjugate with CRCL3 on Day of Use

Preparation of DNP-KLH, HGG

Preparation of KLH

Preparation of KLH, HGG and Coated Gluteraldehyde Fixed SRBC for Hemoglutination

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

TNP Conjugation to SRBC for Hemoglutination and Plaquing

Use of Biotin-Activated Esters for Protein Comjugation

Pronase Stripping of Membrane Immunoglobin in Rabbits

Cell & FACS Staining

Autoradiography of Cell Smears

Chemical Identification of Monocytes

Cytochemical Identification of Monocytes and Granulocytes (article)

Cytoplasmic Fluorescent Antibody Staining

F-KLH Staining Procedure

FDA Vital Cell Stain

Human Antibody Titration Protocol

ICN Biomedicals

Mario's Conjugated Protocols

Mario's Perspective on Compensation

Membrane Fluorescent Antibody Staining

Microtier Staining Method

Mithramycin Staining Protocol

Standard FACS Surface Staining Protocol

Stripping of Surface Globulin in Mouse Cells

Titration Analysis Protocol

Titration Analysis Worksheet

Wright Giesma Stain

Wright Giesma Stain (Blood Smears)

Cell Preparation

AO/EB

Bone Marrow Removal Technique

CBRC

FDA

LeucoPrep from BD (PDF)

Dissection of Mouse Spleen

Cell Treatment with Glass Wool

Preparation of Buffy Coat

Counting Cells

Counting Cells with an Electric Counter

Loading Cells onto Gradient

Separation of Lymphocytes Using Ficoll-Hypaque

Ficoll-Isopaque Technique

Method doe Velocity Sedimentation Ig Gradient

Identification of Dead Cells in FACS Cloning Procedure

A Solid-Phase Immunoenzymatic Technique for the Enumeration of Specific Antibody-Secreting Cells

Culture of Lymph and Spleen

Mitogen Simulation

Mixed Lymphocyte Reaction

Preparation of Human Leukocyte Cultures for Chromosome Analysis

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

Fixation of Cells

Fixation of Cells with Glutaraldehyde

Formlin Fixation of Cells

Labeling

Fluoresceination

Fluoresceinating C ---Valin A

FTC Labeling in Suspension

Radiolabeling

I-125 Labeling of Proteins

Immunoprecipitation of radiolabelled Proteins from NP-40 Extracts

Labeling with I-125

Preparation of Iodogen Tubes

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

Solid Phase I-125 Labeling Protocol Page

TCA Precipitation for Determination of Radioactivity

Separation of T and B Cells

Anti-Theta Treatment of Cells

Dupont Procedure for Wood Columns

Hepes Buffer

Human White Cell Separation

Preparations for  Cell Separators

Procedure for  Cell Separation by Nylon Wool Columns

Solutions

Acid Citrate Dextrose Solution

Antibiotics for Irradiated Mice

Balanced Salt Solution (BSS)

Cell Counting Solutions

Dulbecco Phosphate-Buffered Saline (PBS)

Ethylenediamine Tetraacetic Acid (EDTA or Versene) pH 7.2

Hepes Buffer

Preparation of Concentrated Pen-Strep for Frozen Aliquots

Preparation of Glutamine for Tissue Culture Media

Procedure for IsoMEM Preparation

Sodium Azide NaN3

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

Trypsinization Reagent