Catalog # EP5131

EB3 (Ser-176), phospho-specific Antibody

Rabbit Polyclonal

Application / Dilution

ELISA	1:1000
WB	1:500

Size 100 μl

Species Reactivity Hu, Rt, Ms

MW 32 kDa

Microtubles (MTs) are oriented with a fast growing plus-end and a slower growing minus-end. The MT plus-end is a crucial site for the regulation of MT dynamics and MT association with different cellular organelles by several groups of plus-end tracking proteins (+TIPs). These +TIPs form comet-like accumulations at the plus ends of MTs to regulate MT dynamics and interactions. The End-Binding (EB) family of +TIPs includes EB1 (MAPRE1), EB2 (MAPRE2, RP1), and EB3 (MAPRE3, EBF3). EB proteins are ubiquitiously expressed +TIPs that can dimerize through a coiled-coil C-terminal region, and bind MTs through an N-terminal calponin homology domain. EB proteins can stabilize MTs, increase MT dynamics, and suppress MT pauses. Site specific phosphorylation may regulate EB activity. EB3 Ser-162 phosphorylation destabilizes EB3 dimer and reduces MT growth, while aurora-kinase induced Ser-176 phosphorylation regulates EB3 protein stability during mitosis.

References

Komarova, Y. et al. (2012) Mol Cell. 48:914.
Gouveia, S.M. & Akhmanova, A. (2010) Int Rev Cell Mol Biol. 285:1.
Komarova, Y. et al. (2005) Mol Biol Cell. 16(11):5334.

Western blot analysis of adult mouse brain untreated (lanes 1, 3, 5, & 7) or treated with lambda phosphatase (lanes 2, 4, 6, & 8). The blots were probed with rat monoclonal anti-EB3 (EM5091) (lanes 1 & 2), and rabbit polyclonals anti-EB3 (Ser-162) (lanes 3 & 4), anti-EB3 (Ser-176) (lanes 5 & 6), and anti-EB3 (a.a. 171-182) (lanes 7 & 8).

Phospho-EB3 (Ser-176) synthetic peptide (coupled to KLH) corresponding to amino acid residues surrounding serine 176 in human EB3. This peptide sequence is highly conserved in mouse and rat EB3. This site is conserved in EB2 but the neighboring amino acids are not well conserved.

*For more information, see UniProt Accession Q9UPY8

Rabbit polyclonal, affinity-purified antibody is supplied in 100µl phosphate-buffered saline, 50% glycerol, 1 mg/ml BSA, and 0.05% sodium azide. Store at –20°C. Stable for 1 year.

The products are are safely shipped at ambient temperature for both domestic and international shipments. Each product is guaranteed to match the specifications as indicated on the corresponding technical data sheet. Please store at -20C upon arrival for long term storage.

This antibody was cross-absorbed to unphospho-EB3 (Ser-176), before affinity purification using phospho-EB3 (Ser-176) peptide (without carrier). The antibody detects a 32 kDa* protein corresponding to the apparent molecular mass of phosphorylated EB3 on SDS-PAGE immunoblots of mouse brain. This reactivity is significantly reduced after lambda phosphatase treatment.

*All molecular weights (MW) are confirmed by comparison to Bio-Rad Rainbow Markers and to western blot mobilities of known proteins with similar MW.

This kit contains: