Anti-phospho AKAP-12/Gravin Antibody (Thr766) (C15-1317-688)

General description

A-kinase anchor protein 12 (UniProt Q02952; also known as A-kinase anchor protein 250 kDa, AKAP-12, AKAP 250, Gravin, Myasthenia gravis autoantigen, SSeCKS) is encoded by the AKAP12 (also known as AKAP250) gene (Gene ID 9590) in human. AKAP-12/Gravin is a member of the AKAPs (A-kinase anchoring proteins) family of scaffold proteins that are characterized by a binding domain for the RI/RII regulatory subunit of protein kinase A (a.a. 1541-1554 of AKAP-12) and play critical roles in establishing spatial constraints for multivalent signalling assemblies. AKAP-12/Gravin form a scaffold with PKA and the mitotic kinases, Aurora A and Plk1, at the mother spindle pole during metaphase, guarding against errors occurring during cell division by constraining active Aurora A and Plk1 at the end of the spindle to facilitate signal relay from one kinase to the next. AKAP-12/Gravin is phosphorylated on Thr766 in mitotic cells, T766A phosphosite mutant is unable to interact with Plk1 and negatively impacts cell proliferation. AKAP-12/Gravin is reported to be depleted in proliferating germ line-derived tumors from patients with testicular seminoma, and seminiferous tubular cells divide more rapidly in Akap12-knockout mice. Mutation or amplification of AKAP12 has also been linked to melanoma, prostate, and ovarian cancers in human.

Specificity

The Thr766 numbering is based on the mature sequence (Met1 removed) of human Gravin isoform 1 (Alpha), which is equivalent to Thr668 of isoform 2 (Beta) and Thr661 of isoform 3 (Gamma).

Immunogen

Epitope: pThr766.

KLH-conjugated linear peptide corresponding to a human AKAP-12/Gravin sequence containing phosphorylated Thr766.

Application

Immunocytochemistry Analysis: A representative lot detected an asymmetric enrichment of AKAP-12/Gravin pThr766 immunoreactivity at the mother spindle poles over the daughter spindle poles by fluorescent immunocytochemistry staining of fixed metaphase MEFs and prometaphase HEK293 cells (Hehnly, H., et al. (2015). Elife. 4. pii: e09384).
Immunocytochemistry Analysis: A representative lot detected AKAP-12/Gravin Thr766 phosphorylation in mitotic HEK293 and RAT-2 cells (from prophase through telophase/cytokinesis), but not cells in interphase, by fluorescent immunocytochemistry staining of methanol-fixed and permeabilized cells (Canton, D.A., et al. (2012). Mol.Cell. 48(4):547-559).
Western Blotting Analysis: A representative lot detected AKAP-12/Gravin Thr766 phosphorylation in HEK293 cell lysate (Hehnly, H., et al. (2015). Elife. 4. pii: e09384).

Research Category
Signaling

Research Sub Category
Cell Cycle, DNA Replication & Repair

This Anti-phospho AKAP-12/Gravin Antibody (Thr766) is validated for use in Immunocytochemistry, Western Blotting for the detection of phospho AKAP-12/Gravin.

Quality

Evaluated by Immunocytochemistry in HeLa cells.

Immunocytochemistry Analysis: A 1:500 dilution of this antibody detected AKAP-12/Gravin Thr766 phosphorylation in HeLa cells.

Target description

191.4 kDa (isoform 1; Alpha), 181.6 kDa (isoform 2; Beta), 180.9 kDa (isoform 3; Gamma) calculated (Met1 removed). Due to posttranslational phosphorylation on multiple sites, target band size of greater than 250 kDa was reported (Hehnly, H., et al. (2015). Elife. 4. pii: e09384; Canton, D.A., et al. (2012). Mol.Cell. 48(4):547-559).

Physical form

Rabbit polyclonal antibody serum with 0.05% sodium azide.

Unpurified.

Storage and Stability

Stable for 1 year at -20°C from date of receipt.
Handling Recommendations: Upon receipt and prior to removing the cap, centrifuge the vial and gently mix the solution. Aliquot into microcentrifuge tubes and store at -20°C. Avoid repeated freeze/thaw cycles, which may damage IgG and affect product performance.

Other Notes

Concentration: Please refer to lot specific datasheet.

Disclaimer

Unless otherwise stated in our catalog or other company documentation accompanying the product(s), our products are intended for research use only and are not to be used for any other purpose, which includes but is not limited to, unauthorized commercial uses, in vitro diagnostic uses, ex vivo or in vivo therapeutic uses or any type of consumption or application to humans or animals.

biological source: rabbit. Quality Level: 100. antibody form: serum. antibody product type: primary antibodies. clone: polyclonal. species reactivity: rat, human. species reactivity (predicted by homology): rhesus macaque (based on 100% sequence homology). technique(s): immunocytochemistry: suitable, western blot: suitable. NCBI accession no.: NP_005091. UniProt accession no.: Q02952. shipped in: dry ice. target post-translational modification: phosphorylation (pThr766). Gene Information: human ... AKAP12(9590). Storage Class Code: 12 - Non Combustible Liquids. WGK: WGK 1. Flash Point(F): Not applicable. Flash Point(C): Not applicable.