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.
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.
Preparation and Storage
Sterile PBS to a final concentration of 0.2 mg/mL.
Reconstitution Buffer Available
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.
Calcium/Calmodulin-dependent Protein Kinase I
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 220.127.116.11; MGC120317; MGC120318
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