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Purified Mouse Anti-CaM Kinase II
Purified Mouse Anti-CaM Kinase II

Western blot analysis of CaM Kinase II expression on a rat cerebrum lysate (left panel). Rat Cerebrum Lysate (Cat. No. 611463) was stained with Purified Mouse Anti-CaM Kinase II (Cat. No. 611292/611293) at dilutions of 1: 1:2500, 1:5000, and 1:10,000 (lanes 1, 2, and 3 respectively). CaM Kinase II expression was visualized with HRP Goat Anti-Mouse Ig (Cat. No. 554002) and appears as a 52 kDa band.

Immunohistochemical analysis of CaM Kinase II expression on rat brain (center panel). Formalin-fixed paraffin-embedded section without citrate buffer pretreatment (10X magnification).

Immunofluorescence analysis of CaM Kinase II expression on SK-N-SH cells (Human neuroblastoma; ATCC HTB-11) (right panel).  Cells were seeded in a collagen coated 384-well imaging plate at ~ 8,000 cells per well.  After overnight incubation, cells were stained using the methanol fix/perm protocol and Purified Mouse Anti-CaM Kinase II.  The second step reagent used was Alexa Fluor® 488 goat anti-mouse Ig (Invitrogen) (pseudo colored green). Cell nuclei were counter stained with Hoechst 33342 (pseudo colored blue). Images was taken either on a BD Pathway™ 855 or 435 Bioimager System with a 20x objective and merged using BD AttoVision™ software. This antibody also stains SH-SY5Y, C6, U87 and U373 cells using both the Triton X-100 and methanol fix/perm protocols.

Western blot analysis of CaM Kinase II expression on a rat cerebrum lysate (left panel). Rat Cerebrum Lysate (Cat. No. 611463) was stained with Purified Mouse Anti-CaM Kinase II (Cat. No. 611292/611293) at dilutions of 1: 1:2500, 1:5000, and 1:10,000 (lanes 1, 2, and 3 respectively). CaM Kinase II expression was visualized with HRP Goat Anti-Mouse Ig (Cat. No. 554002) and appears as a 52 kDa band.

Immunohistochemical analysis of CaM Kinase II expression on rat brain (center panel). Formalin-fixed paraffin-embedded section without citrate buffer pretreatment (10X magnification).

Immunofluorescence analysis of CaM Kinase II expression on SK-N-SH cells (Human neuroblastoma; ATCC HTB-11) (right panel).  Cells were seeded in a collagen coated 384-well imaging plate at ~ 8,000 cells per well.  After overnight incubation, cells were stained using the methanol fix/perm protocol and Purified Mouse Anti-CaM Kinase II.  The second step reagent used was Alexa Fluor® 488 goat anti-mouse Ig (Invitrogen) (pseudo colored green). Cell nuclei were counter stained with Hoechst 33342 (pseudo colored blue). Images was taken either on a BD Pathway™ 855 or 435 Bioimager System with a 20x objective and merged using BD AttoVision™ software. This antibody also stains SH-SY5Y, C6, U87 and U373 cells using both the Triton X-100 and methanol fix/perm protocols.

Product Details
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BD Transduction Laboratories™
Ca2+/calmodulin-dependent protein kinase II
Rat (QC Testing), Human, Mouse, Dog (Tested in Development)
Mouse IgG1
Rat CaM Kinase IIα aa. 448-460
Bioimaging, Western blot (Routinely Tested), Immunofluorescence, Immunohistochemistry (Tested During Development)
52 kDa
250 µg/ml
AB_398819
Aqueous buffered solution containing BSA, glycerol, and ≤0.09% sodium azide.
RUO


Preparation And Storage

The monoclonal antibody was purified from tissue culture supernatant or ascites by affinity chromatography. Store undiluted at -20°C.

Product Notices

  1. Since applications vary, each investigator should titrate the reagent to obtain optimal results.
  2. Caution: Sodium azide yields highly toxic hydrazoic acid under acidic conditions. Dilute azide compounds in running water before discarding to avoid accumulation of potentially explosive deposits in plumbing.
  3. Source of all serum proteins is from USDA inspected abattoirs located in the United States.
  4. Sodium azide is a reversible inhibitor of oxidative metabolism; therefore, antibody preparations containing this preservative agent must not be used in cell cultures nor injected into animals. Sodium azide may be removed by washing stained cells or plate-bound antibody or dialyzing soluble antibody in sodium azide-free buffer. Since endotoxin may also affect the results of functional studies, we recommend the NA/LE (No Azide/Low Endotoxin) antibody format, if available, for in vitro and in vivo use.
  5. Please refer to http://regdocs.bd.com to access safety data sheets (SDS).
  6. Species cross-reactivity detected in product development may not have been confirmed on every format and/or application.
  7. Triton is a trademark of the Dow Chemical Company.
  8. Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
611292 Rev. 2
Antibody Details
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45/CaM Kinase II

Ca2+/calmodulin-dependent protein kinase II (CaM kinase II) is a multifunctional Ser/Thr kinase that regulates a number of cellular functions in response to increased intracellular Ca2+. CaM kinase II is widely distributed, but is predominantly expressed in brain. It is involved in the regulation of neuronal functions such as neurotransmitter synthesis, neurotransmitter release, long-term potentiation, and formation of spatial learning. Neuronal CaM kinase II contains heteromers of two major subunits, α and β, at a ratio of 2:1 and homomers of α subunits. Each subunit has N-terminal ATP-binding and catalytic/regulatory domains and a C-terminal association domain. The regulatory domain consists of the autoinhibitory and calmodulin-binding sites. Assembly of the association domains of multiple subunits positions the regulatory domains for intersubunit autophosphorylation. After binding Ca2+/calmodulin, CaM kinase II undergoes rapid autophosphorylation of the α and β subunits, which results in a substantial increase in its affinity for Ca2+/calmodulin.

611292 Rev. 2
Format Details
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Purified
Tissue culture supernatant is purified by either protein A/G or affinity purification methods. Both methods yield antibody in solution that is free of most other soluble proteins, lipids, etc. This format provides pure antibody that is suitable for a number of downstream applications including: secondary labeling for flow cytometry or microscopy, ELISA, Western blot, etc.
Purified
611292 Rev.2
Citations & References
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Development References (5)

  1. Brocke L, Chiang LW, Wagner PD, Schulman H. Functional implications of the subunit composition of neuronal CaM kinase II. J Biol Chem. 1999; 274(32):22713-22722. (Biology). View Reference
  2. Fallon L, Moreau F, Croft BG, Labib N, Gu WJ, Fon EA. Parkin and CASK/LIN-2 associate via a PDZ-mediated interaction and are co-localized in lipid rafts and postsynaptic densities in brain. J Biol Chem. 2002; 277(1):486-491. (Biology: Western blot). View Reference
  3. Hanson PI, Schulman H. Neuronal Ca2+/calmodulin-dependent protein kinases. Annu Rev Biochem. 1992; 61:559-601. (Biology). View Reference
  4. Ishida A, Fujisawa H. Stabilization of calmodulin-dependent protein kinase II through the autoinhibitory domain. J Biol Chem. 1995; 270(5):2163-2170. (Biology). View Reference
  5. Zong H, Ren JM, Young LH, et al. AMP kinase is required for mitochondrial biogenesis in skeletal muscle in response to chronic energy deprivation. Proc Natl Acad Sci U S A. 2002; 99(25):15983-15987. (Biology: Western blot). View Reference
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611292 Rev. 2

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Comparisons, where applicable, are made against older BD Technology, manual methods or are general performance claims.  Comparisons are not made against non-BD technologies, unless otherwise noted.

For Research Use Only. Not for use in diagnostic or therapeutic procedures.