Human CaM Kinase I Antibody

Catalog #: AF7899 Datasheet / COA / SDS

Catalog #	Availability	Size / Price	Qty
AF7899
AF7899-SP

Detection of Human CaM Kinase I by Western Blot.

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All CaM Kinase I Antibodies

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Summary Data Examples Preparation and Storage Reconstitution Calculator Background Product Datasheets Related Research Areas

Human CaM Kinase I Antibody Summary

Species Reactivity

Human

Specificity

Detects recombinant human CaM Kinase I alpha in direct ELISAs and Western blots. In direct ELISAs, less than 1% cross-reactivity with recombinant human CaMKIV is observed.

Source

Polyclonal Sheep IgG

Purification

Antigen Affinity-purified

Immunogen

E. coli-derived recombinant human CaM Kinase I alpha
Ser176-Arg254 (Ala224Ser)
Accession # Q14012

Formulation

Lyophilized from a 0.2 μm filtered solution in PBS with Trehalose. *Small pack size (SP) is supplied either lyophilized or as a 0.2 µm filtered solution in PBS.

Applications

Recommended Concentration

Sample

Western Blot

2 µg/mL

See below

Immunohistochemistry

5-15 µg/mL

See below

Please Note: Optimal dilutions should be determined by each laboratory for each application. General Protocols are available in the Technical Information section on our website.

Scientific Data

Western Blot

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Detection of Human CaM Kinase I by Western Blot. Western blot shows lysates of IMR-32 human neuroblastoma cell line, U-118-MG human glioblastoma/astrocytoma cell line, and human brain (cortex) tissue. PVDF membrane was probed with 2 µg/mL of Sheep Anti-Human CaM Kinase I Antigen Affinity-purified Polyclonal Antibody (Catalog # AF7899) followed by HRP-conjugated Anti-Sheep IgG Secondary Antibody (Catalog # HAF016). A specific band was detected for CaM Kinase I at approximately 37 kDa (as indicated). This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 1.

Immunohistochemistry

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CaM Kinase I alpha in Human Hypothalamus. CaM Kinase I was detected in immersion fixed paraffin-embedded sections of human hypothalamus using Sheep Anti-Human CaM Kinase I Antigen Affinity-purified Polyclonal Antibody (Catalog # AF7899) at 1.7 µg/mL overnight at 4 °C. Tissue was stained using the Anti-Sheep HRP-DAB Cell & Tissue Staining Kit (brown; Catalog # CTS019) and counterstained with hematoxylin (blue). Specific staining was localized to cytoplasm and nuclei. View our protocol for Chromogenic IHC Staining of Paraffin-embedded Tissue Sections.

Reconstitution Calculator

Preparation and Storage

Reconstitution

Sterile PBS to a final concentration of 0.2 mg/mL.

Shipping

The product is shipped at ambient temperature. Upon receipt, store it immediately at the temperature recommended below. *Small pack size (SP) is shipped with polar packs. Upon receipt, store it immediately at -20 to -70 °C

Stability & Storage

Use a manual defrost freezer and avoid repeated freeze-thaw cycles.

12 months from date of receipt, -20 to -70 °C as supplied.
1 month, 2 to 8 °C under sterile conditions after reconstitution.
6 months, -20 to -70 °C under sterile conditions after reconstitution.

Background: CaM Kinase I

CaMKI (Calcium/Calmodulin-dependent kinase I) is a 37-43 kDa monomeric and cytosolic member of the CaMK subfamily, Ser/Thr protein kinase family, protein kinase superfamily of molecules. It is found in multiple cell types, including adrenal cortical cells, neurons and macrophages/osteoclast precursors. CaMKI is associated with the cellular Ca⁺⁺ signaling pathway. Upon entry into the cell, calcium levels are sensed by calmodulin. Upon Ca⁺⁺ binding, calmodulin (and CaMK kinase) interacts with a Ser/Thr protein kinase termed CaMKIa. CaMKIa is of particular interest in neurons and cells of the zona glomerulosa. In neurons, CaMKIa activation results in the recruitment of AMPA-Rs and the promotion of axon growth (vs.dendrite outgrowth which is promoted by CaMKIg). In the adrenal gland, CaMKIa regulates CYP11B2 transcription, and thus the ability to synthesize aldosterone. Human CaMKI, the isoform used for immunization, is 370 amino acids (aa) in length (SwissProt #:Q14012). It contains one protein kinase domain (aa 20-276) followed by a calmodulin-binding region (aa 296-317) and an NES (aa 315-321). Over aa 178-257, human and mouse CaMKI are identical in amino acid sequence. There are three additional CaMKI isoforms (b, g, d), all the product of distinct genes. Nevertheless, over the same aa sequence, these three isoforms (b, g, d) share 80%, 81% and 97% aa sequence identity with CaMKI(a), respectively.

Long Name

Calcium/Calmodulin-dependent Protein Kinase I

Alternate Names

calcium/calmodulin-dependent protein kinase I; calcium/calmodulin-dependent protein kinase type 1; CaM kinase I alpha; CaM Kinase I; CAMKI; caM-KI; caMKI-alpha; EC 2.7.11; EC 2.7.11.17; MGC120317; MGC120318