Invitrogen

Phospho-CaMKII alpha (Thr286) Monoclonal Antibody (22B1)

Product Image Gallery

Phospho-CaMKII alpha (Thr286) Antibody — Published figure using Phospho-CaMKII alpha (Thr286) monoclonal antibody (Product # MA1-047)

Phospho-CaMKII alpha (Thr286) Antibody in Immunofluorescence (IF) — Immunofluorescent analysis of Phospho-CaM Kinase II pThr286 in Neuro-2a cells (left panel) or cells stimulated with Lamda-Phosphatase (Lambda-PPase) (right panel). Formalin-fixed cells were permeabilized with 0.1% Triton X-100 in TBS for 5-10 minutes at room temperature and blocked with 3% BSA-PBS for 30 minutes at room temperature. Cells were probed with a Phospho-CaM Kinase II pThr286 Monclonal Antibody (22B1) (Product # MA1-047) at a dilution of 1:50 and incubated overnight in a humidified chamber. Cells were washed with PBST and incubated with a DyLight-conjugated secondary antibody for 45 minutes at room temperature in the dark. F-actin (red) was stained with a fluorescent phalloidin and nuclei (blue) were stained with DAPI. Images were taken at a 60X magnification.

Phospho-CaMKII alpha (Thr286) Antibody in Western Blot (WB) — Western blot detection of unphosphorylated and phosphorylated rat brain homogenate (left and right lanes respectively) using Product # MA1-047 and Product # MA1-048 antibodies.

Product Details

Tested Applications

Dilution

ELISA (ELISA)

Assay dependent

Flow Cytometry (Flow)

1 µg / 10^6 cells

Immunocytochemistry (ICC)

1:100

Immunofluorescence (IF)

1:50

Immunohistochemistry (IHC)

Assay dependent

Immunoprecipitation (IP)

Assay dependent

Western Blot (WB)

1:2,000

Published Applications

Immunofluorescence (IF)

See 1 publication below

Western Blot (WB)

See 50 publications below

Miscellaneous PubMed (MISC)

See 1 publication below

Immunohistochemistry (IHC)

See 5 publications below

Immunocytochemistry (ICC)

See 2 publications below

Product Specifications

Species Reactivity

Human, Mouse, Rat

Published species

Human, Mouse, Rabbit, Rat

Host / Isotype

Mouse / IgG1

Class

Monoclonal

Type

Antibody

Clone

22B1

Immunogen

Synthetic peptide corresponding to residues M(281) H R Q E T(p) V D C L K K F N(294) of rat CaM Kinase II protein.

Conjugate

Unconjugated

Form

Liquid

Concentration

1 mg/mL

Purification

Protein G

Storage buffer

PBS

Contains

0.05% sodium azide

Storage conditions

-20° C, Avoid Freeze/Thaw Cycles

RRID

AB_325402

Product Specific Information

MA1-047 detects phosphorylated CaM kinase II from human, rat and mouse tissues/cells. This antibody has been shown to detect the phosphorylated alpha subunit CaM kinase II only. MA1-047 does not detect the non-phosphorylated alpha subunit CaM kinase II nor with the ~60 kDa beta subunit in either phosphorylation state.

MA1-047 has been successfully used in Western blot, immunofluorescence, immunoprecipitation, immunocytochemistry, immunohistochemistry, and ELISA procedures. By Western blot, this antibody detects an ~50 kDa band representing the alpha subunit of CaM kinase II only after phosphorylation. Immunofluorescence staining of CaM kinase II in rat/mouse hippocampal cells with MA1-047 densely labels cell somas and neuropil areas, while nuclei are only lightly stained.

The MA1-047 immunizing peptide is a synthetic peptide corresponding to residues M(281) H R Q E T(p) V D C L K K F N(294) of rat CaM Kinase II protein with a phosphorylated threonine at position 286.

Target Information

CaM kinase II, in a calcium and calmodulin dependent manner, phosphorylates many different brain substrates including synapsin I, tryptophan hydroxylase, tyrosine hydroxylase and nitric oxide synthase thereby performing regulatory functions associated with increases in intracellular free calcium. CaM kinase II is particularly abundant in the hippocampus and forebrain where it comprises ~1% of total protein. Within the neuron, CaM kinase II is a major component of the postsynaptic density fraction accounting for as much as 30% of total protein in the post synaptic density (PSD) region.

Autophosphorylation of threonine-286 occurs after calcium/calmodulin activation and can enable CaM kinase II to remain active independent of free calcium. Autophosphorylation of CaM kinase II in hippocampal neurons, which results in relatively high levels of kinase activity even at basal calcium concentrations, may allow for both upward and downward regulation as opposed to simply on/off regulation.

For Research Use Only. Not for use in diagnostic procedures. Not for resale without express authorization.

Bioinformatics

Protein Aliases: alpha CaM kinase II; alpha-CaMKII; Ca2+/calmodulin-dependent protein kinase II alpha; calcium/calmodulin-dependent protein kinase (CaM kinase) II alpha; calcium/calmodulin-dependent protein kinase II alpha subunit; calcium/calmodulin-dependent protein kinase II alpha-B subunit; Calcium/calmodulin-dependent protein kinase type 2; calcium/calmodulin-dependent protein kinase type II alpha chain; Calcium/calmodulin-dependent protein kinase type II subunit alpha; CaM kinase II alpha subunit; CaM kinase II subunit alpha; CaM-kinase II alpha chain; CaMK II; CaMK-II alpha subunit; caMK-II subunit alpha; CaMKIINalpha; Phospho-CaMKI

Gene Aliases: CAMK2A; CAMKA; CaMKII; KIAA0968; mKIAA0968; PK2CDD; PKCCD; R74975

UniProt ID: (Human) Q9UL21, (Mouse) P11798, (Rat) P11275

Entrez Gene ID: (Human) 815, (Mouse) 12322, (Rat) 25400

Molecular Function: non-receptor serine/threonine protein kinase