CaMKII (Alpha-Specific) Antibody (OASE00036)

• Product Number: OASE00036
• Name: CaMKII (Alpha-Specific) Antibody (OASE00036)
• Clone: 6G9
• Isotype: IgG1
• Conjugation: Unconjugated
• NCBI Gene Id: 12322
• Host: Mouse
• Clonality: Monoclonal
• Concentration: 1 mg/ml
• Gene Full Name: calcium/calmodulin-dependent protein kinase II alpha
• Description: CaMKII is an important member of the calcium/calmodulin-activated protein kinase family, functioning in neural synaptic stimulation and T-cell receptor signaling (1, 2). CaMKII is expressed in many different tissues but is specifically found in the neurons of the forebrain and its mRNA is found within the dendrites and the soma of the neuron. The CaMKII that is found in the neurons consist of two subunits of 52 (termed alpha genes) and 60 kDa (beta genes). CaMKII has catalytic and regulatory domains, as well as an ATP-binding domain, and a consensus phosphorylation site (3-7). The binding of Ca2+/calmodulin to its regulatory domain releases its auto inhibitory effect and activates the kinase (8). This kinase activation results in autophosphorylation at threonine 286 (8). The threonine phosphorylation state of CaMKII can be regulated through PP1/PKA. Whereas PP1 (protein phosphatase 1) dephosphorylates phospho-CaMKII at Thr286, PKA (protein kinase A) prevents this dephosphorylation (9). Autophosphorylation also enables CaMKII to attain an enhanced affinity for NMDA receptors in postsynaptic densities (10-12).
• Alias Symbols: CamK2, CamK2A, CamK2B, CamK2D, CamK2G, CAMKA
• Product Format: Liquid PBS (pH 7.4)
• Reference: 1. Hughes K. et al. (2001) J. Biol. Chem. 276: 36008–36013. 2. Barria A. et al. (1997) Science 276: 2042–2045.
  3. Bennet M.K. and Kennedy M.B. (1987) Proc. Natl. Acad. Sci. U.S.A. 84: 1794-1798.
  4. Broke L., Srinivasan M. and Schulman H. (1995) J. Neurosci. 15: 6797-6808.
  5. Nghiem P., Saati S. M., Martens C. L., Gardner P. and Schulman H. (1993) J. Biol. Chem. 268: 5471-5479.
  6. Edman C.F. and Schulman H. (1994) Biochem. Biophys. Acta 1221: 90-102.
  7. Tombes R.M. and Krystal G.W., (1997) Biochem. Biophys. Acta 13555: 281-292.
  8. Means A.R. (2000) Mol. Endocrinol. 14: 4–12.
  9. Makhinson M. et al. (1999) J. Neurosci. 19: 2500–2510.
  10. Strack S. and Colbran R.J. (1998) J. Biol. Chem. 273: 20689-20692.
  11. Leonard S.A., Lim I.A., Hemsworth D.E., Horne M.C. and Hell J.W. (1999) Proc. Natl. Acad. Sci. U.S.A. 96: 3239-3244.
  12. Shen K. and Meyer Y. (1999) Science 284: 162-167.
• Description of Target: CaMKII is an important member of the calcium/calmodulin-activated protein kinase family, functioning in neural synaptic stimulation and T-cell receptor signaling (1, 2). CaMKII is expressed in many different tissues but is specifically found in the neurons of the forebrain and its mRNA is found within the dendrites and the soma of the neuron. The CaMKII that is found in the neurons consist of two subunits of 52 (termed alpha genes) and 60 kDa (beta genes). CaMKII has catalytic and regulatory domains, as well as an ATP-binding domain, and a consensus phosphorylation site (3-7). The binding of Ca2+/calmodulin to its regulatory domain releases its auto inhibitory effect and activates the kinase (8). This kinase activation results in autophosphorylation at threonine 286 (8). The threonine phosphorylation state of CaMKII can be regulated through PP1/PKA. Whereas PP1 (protein phosphatase 1) dephosphorylates phospho-CaMKII at Thr286, PKA (protein kinase A) prevents this dephosphorylation (9). Autophosphorylation also enables CaMKII to attain an enhanced affinity for NMDA receptors in postsynaptic densities (10-12).
• Reconstitution and Storage: Store at -20°C for 1 year. Avoid freeze/thaw cycles.
• Specificity: Detects ~50-60kDa. Recognizes both phosphorylated and non-phosphorylated forms.
• Immunogen: Partially purified rat CaMKII
• Additional Information: Certificate of Analysis: 0.1ug/ml was sufficient for detection of CamKII in 20ug rat brain tissue extract by colorimetric immunoblot analysis using Goat Anti-Mouse IgG:AP as the secondary.
• Dilution: WB (1:10000), IHC (1:2000), ICC/IF (1:50); optimal dilutions for assays should be determined by the user.
• Storage Buffer: PBS pH7.4, 50% glycerol, 0.09% sodium azide
• Uniprot ID: P11798
• Protein Accession #: NP_033922.1
• Purification: Protein G Purified
• Gene Symbol: CaMKII
• Predicted Species Reactivity: Human, Mouse, Rat, Bovine
• Application: WB, IHC, ICC, IF, IP, ELISA, RIA
• Image 1: Immunofluorescence
  
  Immunocytochemistry/Immunofluorescence analysis using Mouse Anti-CaMKII Monoclonal Antibody, Clone 6G9. Tissue: HaCaT cells. Species: Human. Fixation: Cold 100% methanol for 10 minutes at -20C. Primary Antibody: Mouse Anti-CaMKII Monoclonal Antibody at 1:100 for 1 hour at RT. Secondary Antibody: FITC Goat Anti-Mouse (green) at 1:50 for 1 hour at RT. Localization: Nuclear Staining.
• Image 2: Immunofluorescence
  
  Immunocytochemistry/Immunofluorescence analysis using Mouse Anti-CaMKII Monoclonal Antibody, Clone 6G9. Tissue: dissociated hippocampal neurons. Species: Mouse. Fixation: Cold 4% paraformaldehyde/0.2% glutaraldehyde in 0.1M sodium phosphate buffer. Primary Antibody: Mouse Anti-CaMKII Monoclonal Antibody at 1:1000 for 12 hours at 4C. Secondary Antibody: FITC Goat Anti-Mouse IgG (green) at 1:50 for 30 minutes at RT. Magnification: 10X. Courtesy of: Mary Kennedy, Caltech.
• Image 3: Immunohistochemistry
  
  Immunohistochemistry analysis using Mouse Anti-CaMKII Monoclonal Antibody, Clone 6G9. Tissue: colon carcinoma. Species: Human. Fixation: Formalin. Primary Antibody: Mouse Anti-CaMKII Monoclonal Antibody at 1:10000 for 12 hours at 4C. Secondary Antibody: Biotin Goat Anti-Mouse at 1:2000 for 1 hour at RT. Counterstain: Mayer Hematoxylin (purple/blue) nuclear stain at 200 ul for 2 minutes at RT. Magnification: 40x.
• Image 4: Western Blot
  
  Western Blot analysis of Rat brain membrane lysate showing detection of CaMKII protein using Mouse Anti-CaMKII Monoclonal Antibody, Clone 6G9. Primary Antibody: Mouse Anti-CaMKII Monoclonal Antibody at 1:1000.