The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
Application notesICC/IF: Use at a concentration of 1 - 2 µg/ml.
IP: 10-20 ug of the antibody can immunoprecipitate EB1 from a cell lysate of HeLa cells.
WB: Use at a concentration of 0.5 - 1 µg/ml. Predicted molecular weight: 30 kDa. Staining of the EB1 band in immunoblotting is specifically inhibited with the immunizing peptide.
Not yet tested in other applications.
Optimal dilutions/concentrations should be determined by the end user.
FunctionMay be involved in microtubule polymerization, and spindle function by stabilizing microtubules and anchoring them at centrosomes. May play a role in cell migration.
Tissue specificityUbiquitously expressed.
Sequence similaritiesBelongs to the MAPRE family. Contains 1 CH (calponin-homology) domain. Contains 1 EB1 C-terminal domain.
DomainComposed of two functionally independent domains. The N-terminal domain forms an hydrophobic cleft involved in microtubule binding and the C-terminal is involved in the formation of mutually exclusive complexes with APC and DCTN1.
Cellular localizationCytoplasm > cytoskeleton. Cytoplasm > cytoskeleton > centrosome. Associated with the microtubule network at the growing distal tip of microtubules. Also enriched at the centrosome.
ab50188 at 1ug/ml dilution staining the “plus-end tracking protein” (+TIP) structures in Rat1 fibroblasts grown to semi-confluency and fixed with acetone/methanol.
The antibody was developed with Goat Anti-Rabbit IgG, Cy3 conjugate. Antibody dilution 1:1,000.
Western blot - MAPRE1 antibody (ab50188)
All lanes : Anti-MAPRE1 antibody (ab50188) at 0.5 µg/ml
Lane 1 : HeLa cell lysate
Lane 2 : NIH3T3 cell lysate
Lane 3 : Rat1 cell lysate
Lane 4 : A431 cell lysate Lane 5 : SW48 cell lysate Lane 6 : HCT116 cell lysate Lane 7 : HEK293 cell lysate
Lane 8 : HeLa cell lysate with EB1 immunizing peptide at 10 µg/ml
Secondary Goat Anti-Rabbit IgG, Peroxidase conjugate at 1/1000 dilution
Shimozawa T et al. Improving spinning disk confocal microscopy by preventing pinhole cross-talk for intravital imaging. Proc Natl Acad Sci U S A110:3399-404 (2013).
Read more (PubMed: 23401517) »
Goldspink DA et al. The microtubule end-binding protein EB2 is a central regulator of microtubule reorganisation in apico-basal epithelial differentiation. J Cell Sci126:4000-14 (2013).
Read more (PubMed: 23813963) »