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Anti-BCKDK Antibody

Rabbit polyclonal antibody to BCKDK

Western blot analysis of BCKDK expression in Hela (A), DLD (B) whole cell lysates.
Immunohistochemical analysis of BCKDK staining in human liver cancer formalin fixed paraffin embedded tissue section. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0). The section was then incubated with the antibody at room temperature and detected using an HRP conjugated compact polymer system. DAB was used as the chromogen. The section was then counterstained with haematoxylin and mounted with DPX.
Immunofluorescent analysis of BCKDK staining in HeLa cells. Formalin-fixed cells were permeabilized with 0.1% Triton X-100 in TBS for 5-10 minutes and blocked with 3% BSA-PBS for 30 minutes at room temperature. Cells were probed with the primary antibody in 3% BSA-PBS and incubated overnight at 4 °C in a humidified chamber. Cells were washed with PBST and incubated with a DyLight 594-conjugated secondary antibody (red) in PBS at room temperature in the dark. DAPI was used to stain the cell nuclei (blue).

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Product Information
Application Applications Legend: E=ELISA WB=Western Blotting IHC=Immunohistochemistry IHC-P=Immunohistochemistry (Paraffin) IP=Immunoprecipitation IF=Immunofluorescence IC=Immunochemistry ICC=Immunocytochemistry FC=Flow Cytometry DB=Dot Blot	WB, IF/IC, IHC
Primary Accession	O14874
Other Accession	O55028
Reactivity	Human, Mouse, Rat, Monkey, Pig, Bovine, Drosophila
Host	Rabbit
Clonality	Polyclonal
Calculated MW	46360 Da

Additional Information
Gene ID	10295
Other Names	[3-methyl-2-oxobutanoate dehydrogenase [lipoamide]] kinase mitochondrial; Branched-chain alpha-ketoacid dehydrogenase kinase; BCKD-kinase; BCKDHKIN
Target/Specificity	Recognizes endogenous levels of BCKDK protein.
Dilution	WB~~WB (1/500 - 1/1000), IHC (1/100 - 1/200), IF/IC (1/100 - 1/500) IF/IC~~N/A IHC~~WB (1/500 - 1/1000), IHC (1/100 - 1/200), IF/IC (1/100 - 1/500)
Format	Liquid in 0.42% Potassium phosphate, 0.87% Sodium chloride, pH 7.3, 30% glycerol, and 0.09% (W/V) sodium azide.
Storage	Store at -20 °C.Stable for 12 months from date of receipt

Protein Information
Name	BCKDK {ECO:0000303\|PubMed:29779826, ECO:0000312\|HGNC:HGNC:16902}
Function	Serine/threonine-protein kinase component of macronutrients metabolism. Forms a functional kinase and phosphatase pair with PPM1K, serving as a metabolic regulatory node that coordinates branched-chain amino acids (BCAAs) with glucose and lipid metabolism via two distinct phosphoprotein targets: mitochondrial BCKDHA subunit of the branched- chain alpha-ketoacid dehydrogenase (BCKDH) complex and cytosolic ACLY, a lipogenic enzyme of Krebs cycle (PubMed:24449431, PubMed:29779826, PubMed:37558654). Phosphorylates and inactivates mitochondrial BCKDH complex a multisubunit complex consisting of three multimeric components each involved in different steps of BCAA catabolism: E1 composed of BCKDHA and BCKDHB, E2 core composed of DBT monomers, and E3 composed of DLD monomers. Associates with the E2 component of BCKDH complex and phosphorylates BCKDHA on Ser-337, leading to conformational changes that interrupt substrate channeling between E1 and E2 and inactivates the BCKDH complex (PubMed:29779826, PubMed:37558654). Phosphorylates ACLY on Ser-455 in response to changes in cellular carbohydrate abundance such as occurs during fasting to feeding metabolic transition. Refeeding stimulates MLXIPL/ChREBP transcription factor, leading to increased BCKDK to PPM1K expression ratio, phosphorylation and activation of ACLY that ultimately results in the generation of malonyl-CoA and oxaloacetate immediate substrates of de novo lipogenesis and glucogenesis, respectively (PubMed:29779826). Recognizes phosphosites having SxxE/D canonical motif (PubMed:29779826).
Cellular Location	Mitochondrion matrix {ECO:0000250\|UniProtKB:Q00972, ECO:0000305\|PubMed:24449431} Note=Detected in the cytosolic compartment of liver cells {ECO:0000250\|UniProtKB:Q00972}
Tissue Location	Ubiquitous.