Bjorkman Lab Protocols

BACTERIAL EXPRESSION

• PI-PLC purification

• Inclusion body preparation

• Ben's E.coli expression and renaturing (also see Ben's website)

BACULOVIRUS

• Baculovirus-transfection

• Sf9 Spinner Media Recipe

Biacore

• Covalent oriented coupling

• Biacore instructions

CALCULATING KD's

• Equilibrium Dialysis

• Scatchard Analysis

• B27 binding assay

CELL SORTING

• Sorting of FcRn expressing cell

CLONING

• Preparation of electocompetent E. coli

• How to label DNA tubes

• PJB's cloning hints

• Cloning tips by Astrid

• Lance's subcloning procedures

• Site-directed mutagenesis protocol

CRYSTALLISATION

• Siliconized coverslips

ELISA

• ELISA

• Anti FcRn ELISA

IMMUNOPRECIPITATION

• Immunoprecipitations (FcRn)

• Immunoprecipitations (FcRn) short

• Immunoprecipitations (HFE)

IMMUNOSTAINING

• Immunostaining of cells

• FcRn sorting

PROTEIN ELECTROPHORESIS

• TCA precipitation protocol

• Silver staining of SDS-PAGE

• SDS-PAGE and Western blot

• Western blot

PROTEIN PURIFICATION

• 1G3 Column-short

• 1G3 Column for FcRn mutants

• Anti-FcRn mAb

• Antibody by Protein G

• HFE - short protocol

• HFE - full protocol

• TFR (his-tagged)

• B27

• FcRn

• Antibodies

• kd/cb2m

• kd/mb2m

• T cell receptor

• UL18 from insect-cell media

• Purification of His-tagged proteins

• nickel vs. cobalt columns

• Fca Purification Protocol

• Concentrator instructions

• Recharging Ni beads

• Osmotic Shock and TRX-htt purification

• Fab production and purification

• PSMA purification

TISSUE CULTURE

• Tissue Culture Guidelines

• Maintenance of HFE producing cells

• Freezing adherent mammalian cells

• Harvesting proteins Chinese Hamster Ovary cells

• Splitting adherent mammalian cells

TRANSFECTION

• Lipofection Procedure

• Ca-P Transfection MDCK

• Stable transfection of CHO-cells

• PCR genes from cells

USDA - APHIS REGULATIONS

• APHIS regulations letter

• U.S. Customs Protein Import

MISC

• Keeping a lab book

• Pichia MeOH induction screening

• BMMY, BMGY recipes

• Metabolic labeling of cells

• Acid elution tips

• Christina's notes on FcRn constructs

• ZipTip

• Preparing figures for publication

• Molecular movies

• Polyclonal Antiserum Production

YUMMIES

• Jenny's Beef Protocol

• Beth's Banana Bread