Lab Methods & Reagents

Capping Protein (CP, CapZ) Reagents
Antibodies at Developmental Studies Hybridoma Bank  DSHB
Antibody name and link Subunit / isoform recognized Species recognized Notes: All work well on immunoblots w/ SDS in sample prep.
mAb 5B12 Alpha1 and alpha2 Chicken, mouse For staining, use glyoxal fixative, e.g. Glyco-Fixx.
mAb 1E5 Beta1, not beta2 Chicken, not mouse or human  
mAb 3F2 Beta2, not beta1 Chicken, mouse, human For staining, use glyoxal fixative, e.g. Glyco-Fixx.
poly Anti-Ysc Cof1 Cofilin Yeast, S. cerevisiae Polyclonal

Expression Plasmids at Addgene

Protein Name Plasmid Expression Vector Addgene Number
CP alpha 1 subunit (Capza1 Mouse) pBJ 2358 Mammalian 89245
CP beta 2 subunit (Capzb Mouse) p-eGFP-beta 2 CP Mammalian 13298
Cadherin 5 (CD144) (CDH5 Human) pBOB-VE-cadherin-GFP Lentiviral 118732
Cadherin 5 (CD144) (CDH5 Human) pBOB-VE-cadherin-tdTomato Lentiviral 118733

Others not listed in the table above (please see Addgene):
  • Budding yeast cofilin (COF1, RFP)
  • Human Lck domain (Lentiviral, tdTomato and GFP)
  • Human CARMIL2 BH Domain (Plasma membrane marker, Lentiviral, tdTomato and GFP)
  • Human Septin 2 (Lentiviral, tdTomato and GFP)
  • Bacterial expression of chicken and mouse CP (wt and mutants)
GFP-Actin Reagent. For vertebrate cells, we use a GFP-actin expression plasmid kindly provided by Dr. Beat Imhof in Geneva. Imhof Lab Web Site.
  • Original paper: Ballestrem et al. (1998). J. Cell Sci. 111, 1649-1658.
  • Example of its use: Merrifield et al. (1999) Nature Cell Biology 1, 72 - 74.

Light Microscopy Methods

Protein Biochemistry Methods

Molecular Biology Methods
  • Gibson Cloning Protocol. From Aaron Kershner and Judith Kimble, Univ Wisconsin - Madison.Based on Gibson DG, Methods Enzymol. 2011. 498:349-61.
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