Catalog # EK6960

EB3 Phospho-Regulation Antibody Kit

Antibody Sampler Kit



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.

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).

Immunocytochemical labeling of EB3 in paraformaldehyde-fixed and NP40-permeabilized A431 cells. The cells were dual labeled with mouse monoclonal anti-α-Tubulin (TM4111) (green) and rat monoclonal anti-EB3 (EM5091) (red). The antibodies were detected using either goat anti-mouse:DyLight® 488 (MS3011) or goat anti-Rat:DyLight® 594 (RS3111).

Rat monoclonal antibodies are supplied in phosphate-buffered saline and 0.05% sodium azide. Rabbit polyclonal antibodies are supplied in phosphate-buffered saline, 50% glycerol, 1 mg/ml BSA, and 0.05% sodium azide. Store all 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.

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

Product References:

EM5101 Komarova, Y. et al. (2005) Mol Biol Cell. 16(11):5334. WB, ICC: CHO-K1 cellls

This kit contains:

EB3 Rat mAb50 μlWB, E, ICCHu, Rt, Ms32
EB3 Rat mAb50 μlWB, E, ICCHu, Rt, Ms32
EB3 (a.a. 171-182) Rabbit pAb50 μlWB, EHu, Rt, Ms32
EB3 (Ser-162), phospho-specific Rabbit pAb50 μlWB, EHu, Rt, Ms32
EB3 (Ser-176), phospho-specific Rabbit pAb50 μlWB, EHu, Rt, Ms32

The end-binding 3 (EB3) phospho-regulation antibody sampler kit can be used to detect the phosphorylation of Ser-162 and Ser-176 in EB3. The kit includes phospho-specific antibodies, as well as rabbit polyclonal and rat monoclonal antibodies for detecting total EB3 expression.