Antibodies to Calmodulin and CaM-kinases - FocusOn 084

Introduction

Calmodulin acts as an intermediary protein that senses calcium levels and relays signals to various calcium-sensitive enzymes, ion channels and other proteins. This 17 kD protein is composed of two globular domains connected together by a flexible linker. Each end binds to two calcium ions. As it binds calcium, Calmodulin undergoes conformational changes which can increase its affinity for target proteins.

Studies have found that Calmodulin participates in the regulation of several biological processes including energy and biosynthetic metabolism, cell motility, exocytosis, cytoskeletal assembly, and intracellular modulation of both cAMP and calcium concentrations. It acts both directly, through interaction with key target enzymes, and indirectly, via specific kinases. Ca2+/Calmodulin (CaM)-dependent protein kinases (CaMkinases). CaM-kinases constitute a family of structurally related enzymes that include phosphorylase kinase, Myosin light chain kinase, and CaM-kinases I-IV.

There have been identified multiple substrates for CaMK I, including synapsin 1, synapsin 2, CREB, and CFTR, and its activation is dependent upon phosphorylation of Thr177 by CaMKkinase (CaMKK). Studies suggest that CaMK I plays an important role in retinal function and its expression is regulated by light stimulation.

Calcium/Calmodulin-dependent protein kinase 1D-beta isoform is a novel Calmodulin-kinase that exhibits Ca2+/CaM-dependent activity. CaMKK alpha upregulates CaMK ID-beta activity through phosphorylation of its Thr180. CaM-kinase II, one of the best-studied multifunctional enzymes, is found in high concentrations in neuronal synapses, and in some regions of the brain it may constitute up to 2% of the total protein content. Activation of CaM-kinase II has been linked to memory and learning processes in the vertebrate nervous system. In the inactive state there is a strong interaction between the inhibitory and catalytic domains of the enzyme. The binding of Ca2+/CaM allows the catalytic domain to phosphorylate the inhibitory domain. Autophosphorylation of Threonine 286 occurs after Ca2+/Calmodulin activation and can enable CaM-Kinase II to remain active independent of free calcium, thereby prolonging the duration of kinase activity.

CaM-kinase IV (CaMK IV), in a calcium and Calmodulin dependent manner, is implicated in phosphorylating numerous transcription factors including the CCAAT enhancer binding protein (C/EBP), serum response factor (SRF), and cyclic AMP response element binding protein (CBP). The nuclear localization of this protein is consistent with its role in mediating calciumdependent gene expression. CaMK IV is particularly abundant in testis, T-cells, and neurons but is also found in other tissues to varying degrees. In neurons, CaMK IV is thought to play an important role in synaptic plasticity via its gene regulatory effects.

Antibody Panel to Calmodulin and CaM-kinases

OriGene Technologies offers a range of monoclonal and polyclonal antibodies to Calmodulin and Calmodulin-Dependent Protein Kinases (CaMKI, II and IV). The antibodies react in various species and can be used for different applications.

For CaM-Kinase II a panel of phospho-specific antibodies is also available. These antibodies have been raised against two different phosphorylation sites: Thr286 and Thr305.

Staining of Calmodulin in rat brain using antibody SM5111.Western blot of CaMK I on At20 cell extract using antibody SP5224P.Western blot of CAMK 1D-beta in human brain cell lysate using antibody SP7340P at 2 µg/ml.Western blot of CaMK IV in rat brain extract using antibody SP5223P.

Fig.1: Staining of Calmodulin in rat brain using antibody SM5111.

Fig.2: Western blot of CaMK I on At20 cell extract using antibody SP5224P.

Fig.3: Western blot of CAMK 1D-beta in human brain cell lysate using antibody SP7340P at 2 µg/ml.

Fig.4: Western blot of CaMK IV in rat brain extract using antibody SP5223P.

Antibody Panel to Calmodulin and CaM-kinases

Calmodulin, CaMK I, CaMK ID-beta, CaMK II antibody.

Literature

Yamanaka A, Hiragami Y, Maeda N, Toku S, Kawahara M, Naito Y, Yamamoto H.Involvement of CaM kinase II in gonadotropin-releasing hormone-induced activation of MAP kinase in cultured hypothalamic neurons.Arch Biochem Biophys. 2007 Oct 15;466(2):234-41.

Kameshita I, Yamada Y, Nishida T, Sugiyama Y, Sueyoshi N, Watanabe A, Asada Y.Involvement of Ca(2+)/calmodulin-dependent protein kinases in mycelial growth of the basidiomycetous mushroom, Coprinus cinereus.Biochim Biophys Acta. 2007 Sep;1770(9):1395-403. Epub 2007 Jun 8

Ishikawa Y, Tokumitsu H, Inuzuka H, Murata-Hori M, Hosoya H, Kobayashi R. Identification and characterization of novel components of a Ca2+/calmodulin-dependent protein kinase cascade in HeLa cells. FEBS Lett. 2003 Aug 28;550(1-3):57-63.

Chin, D. and A.R. Means Mechanisms for regulation of calmodulin kinase IIalpha by Ca2+/calmodulin and autophosphorylation of threonine 286.Biochemistry, 2002, 41(47):14001-14009.

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Primärantikörper

Katalog-Nr. Wirt Iso. Klon Präs. Reakt. Anwendungen  

Calmodulin Antikörper

The cell lysates (40ug) were resolved by SDS-PAGE, transferred to PVDF membrane and probed with anti-human PNMT antibody (1:1000). Proteins were visualized using a goat anti-mouse secondary antibody conjugated to HRP and an ECL detection system. Lane 1.: 293T cell lysate Lane 2.: PNMT transfected 293T cell lysate Mouse IgG2a J4D8 Aff - Purified Hu E, ICC/IF, WB
0.1 ml / 330,00 €
  OriGene Technologies GmbH

Calmodulin Antikörper

The cell lysates (40ug) were resolved by SDS-PAGE, transferred to PVDF membrane and probed with anti-human PNMT antibody (1:1000). Proteins were visualized using a goat anti-mouse secondary antibody conjugated to HRP and an ECL detection system. Lane 1.: 293T cell lysate Lane 2.: PNMT transfected 293T cell lysate Mouse IgG2a J4D8 Aff - Purified Hu E, ICC/IF, WB
50 µl / 220,00 €
  OriGene Technologies GmbH

Calmodulin Antikörper

Calmodulin Rabbit Aff - Purified Hu, Ms, Rt E, P, WB
0.1 mg / 370,00 €
  OriGene Technologies GmbH

Calmodulin (C-term) Antikörper

Formalin-fixed and paraffin-embedded human brain tissue reacted with Calmodulin antibody (C-term), which was peroxidase-conjugated to the secondary antibody, followed by DAB staining. This data demonstrates the use of this antibody for immunohistochemistry; clinical relevance has not been evaluated. Rabbit Ig Purified Hu, Ms, Rt, Ze P, WB
0.4 ml / 415,00 €
  OriGene Technologies GmbH

Calmodulin (C-term) Antikörper

Western blot analysis of CALM1 Antibody (C-term) (CAP17918PU-N) in Hela cell line lysates (35ug/lane). CALM1 (arrow) was detected using the purified Pab.(2ug/ml) Rabbit Hu WB
0.4 ml / 415,00 €
  OriGene Technologies GmbH

CAMK1 (C-term) Antikörper

Formalin-fixed and paraffin-embedded human cancer tissue (hepatocarcinoma) reacted with the primary antibody, which was peroxidase-conjugated to the secondary antibody, followed by DAB staining. This data demonstrates the use of this antibody for immunohistochemistry; clinical relevance has not been evaluated. Rabbit Ig Purified Hu, Ms, Prim, Rt P, WB
0.4 ml / 415,00 €
  OriGene Technologies GmbH

CAMK1D (C-term) Antikörper

Formalin-fixed and paraffin-embedded human cancer tissue (hepatocarcinoma) reacted with the primary antibody, which was peroxidase-conjugated to the secondary antibody, followed by DAB staining. This data demonstrates the use of this antibody for immunohistochemistry; clinical relevance has not been evaluated. Rabbit Ig Purified Hu P, WB
0.4 ml / 415,00 €
  OriGene Technologies GmbH

CAMK1D (N-term) Antikörper

Western blot analysis of CAMK1D (arrow) using rabbit polyclonal CAMK1D Antibody (N-term) (AP17173PU-N). 293 cell lysates (2  µg/lane) either nontransfected (Lane 1) or transiently transfected with the CAMK1D gene (Lane 2). Rabbit Hu WB
0.4 ml / 415,00 €
  OriGene Technologies GmbH

CAMK2A Antikörper

CAMK2A Rabbit Aff - Purified Hu, Ms, Rt WB
0.1 mg / 380,00 €
  OriGene Technologies GmbH

CAMK2A (6-23) Antikörper

Brain, cortex: Formalin-Fixed Paraffin-Embedded (FFPE) Rabbit Purified Can, Hu, Ms, Rt E, ICC/IF, P, WB
50 µg / 425,00 €
  OriGene Technologies GmbH

CAMK2A (C-term) Antikörper

Formalin-fixed and paraffin-embedded human brain tissue reacted with CAMK2 alpha Antibody (C-term) (AP13884PU-N), which was peroxidase-conjugated to the secondary antibody, followed by DAB staining. This data demonstrates the use of this antibody for immunohistochemistry; clinical relevance has not been evaluated. Rabbit Ig Purified Hu P, WB
0.4 ml / 415,00 €
  OriGene Technologies GmbH

CAMK2A (C-term) Antikörper

Formalin-fixed and paraffin-embedded human brain tissue reacted with CAMK2A (C-term), which was peroxidase-conjugated to the secondary antibody, followed by DAB staining Rabbit Ig Purified Hu P, WB
0.4 ml / 415,00 €
  OriGene Technologies GmbH

CAMK2A Antikörper

CAMK2A Rabbit Aff - Purified Hu, Mky, Ms, Rt WB
0.1 ml / 480,00 €
  Neuromics Antibodies

CAMK2A/B Antikörper

Immunofluorescence staining of paraffin-embedded human hippocampal region tissue from the falling sickness disease using CaMKII antibody Rabbit Aff - Purified Hu, Ms, Rt ICC/IF, WB
0.1 mg / 300,00 €
  OriGene Technologies GmbH

CAMK2A/B Antikörper

Immunofluorescence staining of paraffin-embedded human hippocampal region tissue from the falling sickness disease using CaMKII antibody Rabbit Aff - Purified Hu, Ms, Rt ICC/IF, WB
50 µg / 230,00 €
  OriGene Technologies GmbH

CAMK2A/B pThr286 Antikörper

Figure 2. Immunofluorescence staining of paraffin-embedded human hippocampal region tissue from the falling sickness disease using CaMKII (phospho-Thr286) antibody (#AP02526PU, green). Rabbit Aff - Purified Hu, Ms, Rt ICC/IF, WB
0.1 mg / 300,00 €
  OriGene Technologies GmbH

CAMK2A/B pThr286 Antikörper

Figure 2. Immunofluorescence staining of paraffin-embedded human hippocampal region tissue from the falling sickness disease using CaMKII (phospho-Thr286) antibody (#AP02526PU, green). Rabbit Aff - Purified Hu, Ms, Rt ICC/IF, WB
50 µg / 230,00 €
  OriGene Technologies GmbH

CAMK2B (beta/gamma) Antikörper

Western blot (WB) analysis of CaMKIIβ/γ antibody (Cat.-No.: AP06726PU-N) at 1/500 dilution Lane1:HEK293T whole cell lysate Lane2:Raw264.7 whole cell lysate Lane3:Rat colon tissue lysate Rabbit Aff - Purified Hu, Ms, Rt ICC/IF, WB
0.1 mg / 370,00 €
  OriGene Technologies GmbH

CAMK2D (C-term) Antikörper

Formalin-fixed and paraffin-embedded human brain tissue reacted with CAMK2 delta Antibody (C-term) (AP13887PU-N), which was peroxidase-conjugated to the secondary antibody, followed by DAB staining. This data demonstrates the use of this antibody for immunohistochemistry; clinical relevance has not been evaluated. Rabbit Ig Purified Hu, Mky P, WB
0.4 ml / 415,00 €
  OriGene Technologies GmbH

CAMK4 Antikörper

Immunohistochemistry (IHC) analyzes of CaMK4 antibody (Cat.-No.: AP06503PU-N) in paraffin-embedded human breast tissue. Rabbit Aff - Purified Hu, Ms, Rt ICC/IF, P, WB
0.1 mg / 370,00 €
  OriGene Technologies GmbH

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