Product Name:
CaMKK1-pS74
Product Number:
ab-pk557
Target Full Name: Calcium/calmodulin-dependent protein-serine kinase kinase
Target Alias: Calcium/calmodulin-dependent protein kinase 1 alpha, isoform a; Calcium/calmodulin-dependent protein kinase kinase 1, alpha; CaMKK alpha protein; CaMKK1.; CaMKKA; DKFZP761M0423; KKCC1; MGC34095
Product Type Specific: CaMKK1 protein kinase phosphosite-specific antibody
Antibody Code: PK557
Antibody Target Type: Phosphosite-specific
Antibody Phosphosite: S74
Protein UniProt: Q8N5S9
Protein SigNET: CaMKK1
Antibody Type: Polyclonal
Antibody Host Species: Rabbit
Antibody Immunogen Source: Human CaMKK1 (CaMKK) sequence peptide Cat. No.: PE-04AJD99
Antibody Immunogen Sequence: RKL(pS)LQE(bA)C
Antibody Immunogen Description: Corresponds to amino acid residues R71 to E77; in the N-terminal quarter of the protein located before the kinase catalytic domain. S74 phosphorylation stimulates phosphotransferase activity and inhibits phosphotransferase activity by inducing binding to 14-3-3 eta. This is the major in vivo phosphorylation sites in CaMMK1 (≥75 reports from high throughput mass spectrometry studies recorded in PhosphoSitePlus). CaMKK1 is known to be phosphorylated at this site in vitro by PKACa (PRKACA).
Production Method: The immunizing peptide was produced by solid phase synthesis on a multipep peptide synthesizer and purified by reverse-phase hplc chromatography. Purity was assessed by analytical hplc and the amino acid sequence confirmed by mass spectrometry analysis. This peptide was coupled to KLH prior to immunization into rabbits. New Zealand White rabbits were subcutaneously injected with KLH-coupled immunizing peptide every 4 weeks for 4 months. The sera from these animals was applied onto an agarose column to which the immunogen peptide was thio-linked. Antibody was eluted from the column with 0.1 M glycine, pH 2.5. Subsequently, the antibody solution was neutralized to pH 7.0 with saturated Tris.
Antibody Modification: Unconjugated. Contact KInexus if you are interest in having the antibody biotinylated or coupled with fluorescent dyes.
Antibody Concentration: 0.25 / 0.5 mg/ml
Storage Buffer: Phosphate buffered saline pH 7.4, 0.05% Thimerasol
Storage Conditions: For long term storage, keep frozen at -40°C or lower. Stock solution can be kept at +4°C for more than 3 months. Avoid repeated freeze-thaw cycles.
Product Use: Western blotting | Antibody microarray
Antibody Dilution Recommended: 2 µg/ml for immunoblotting
Antibody Species Reactivity: This antibody detects the target phosphoprotein in the following species due to conservation of amino acid sequence: Human | Rhesus Macaque | Dog | Rat | Mouse | Chicken | Frog.
Antibody Positive Control: The observed molecular mass of the processed target protein on SDS-PAGE gels is reported to be around 55-60 kDa.
Antibody Specificity: High-very high
Antibody Cross Reactivity: Weak 37 kDa cross-reactivity detected in phenylarsine oxide (PAO)+vanadate-treated HeLa cells, EGF-treated A431 cells, and insulin-treated MCF7 cells, when these cells were directly homogenized into SDS-PAGE sample buffer.
Related Product 1: CaMKK1 (71-77) pS74 antibody blocking peptide (Cat. No.: PE-04AJD99)
Related Product 2: CaMKK1 expression antibody (Cat. No.: AB-NK022)
Scientific Background: CaMKK1 is a 56 kDa protein-serine/threonine kinase of the Other group and CaMKK family. It is a calcium/calmodulin-activated protein kinase that phosphorylates and activates CaMK1 (at Thr-177), CaMK1d, CaMK1g, CaMK4 (at Thr-196/200), and Akt1. It also targets SAD-B (neuronal development), and SIRT1 as substrates. It promotes cell survival by phosphorylating AKT1/PKB, which inhibits the pro-apoptotic protein BAD. It features an N-terminal catalytic domain and a C-terminal regulatory domain. Binding of calmodulin may releave intrasteric autoinhibition. CaMKK1 is activated by the binding of Ca2+/calmodulin to its regulatory domain, which releases an autoinhibitory segment from the catalytic domain. It is partially inhibited upon phosphorylation by PRCAKA/PKA. It highest expression is in the brain (cerebellum, hippocampus, hypothalamus), but it is expressed in heart and lymphoid tissues. While not crucial for early brain development, it is implicated in regulating neuronal gene transcription and synaptic plasticity. CaMKK1 plays a selective role in long-term contextual fear memory, specifically in males. It is up-regulated in some cancers (lung, hepatocellular carcinoma) and during viral infections (e.g., Cytomegalovirus), where it enhances glycolytic flux. This description may include information annotated by UniProt and/or Google AI.