ANTI-HEXOKINASE 1 CLONE 3B19 ZOOMAB« RABBIT MONOCLONAL (C005B-260949)

General description

ZooMAb^® antibodies represent an entirely new generation of recombinant monoclonal antibodies. Each ZooMAb^® antibody is manufactured using our proprietary recombinant expression system, purified to homogeneity, and precisely dispensed to produce robust and highly reproducible lot-to-lot consistency. Only top-performing clones are released for use by researchers. Each antibody is validated for high specificity and affinity across multiple applications, including its most commonly used application. ZooMAb^® antibodies are reliably available and ready to ship when you need them.

Specificity

Clone 3B19 is a ZooMAb^® rabbit recombinant monoclonal antibody that specifically detects Hexokinase 1. It targets an epitope within 21 amino acids from the N-terminal half.

Immunogen

KLH-conjugated linear peptide corresponding to 21 amino acids from the N-terminal half of human Hexokinase 1.

Application

Quality Control Testing

Evaluated by Western Blotting in Mouse brain tissue lysate.

Western Blotting Analysis: A 1:1,000 dilution of this antibody detected Hexokinase 1 in Mouse brain tissue lysate.

Tested Applications

Western Blotting Analysis: A 1:1,000 dilution from a representative lot detected Hexokinase 1 in lysates from HeLa cells, MCF-7 cells and Rat brain tissue.

Immunohistochemistry (Paraffin) Analysis: A 1:100 dilution from a representative lot detected Hexokinase 1 in Human kidney tissue sections.

Immunocytochemistry Analysis: A 1:100 dilution from a representative lot detected Hexokinase 1 in HeLa cells.

Affinity Binding Assay: A representative lot of this antibody bound Hexokinase 1 peptide with a KD of 2.1 x 10-9 in an affinity binding assay.

Note: Actual optimal working dilutions must be determined by end user as specimens, and experimental conditions may vary with the end user.

Target description

Hexokinase-1 (UniProt: P19367; also known as EC:2.7.1.1, Brain form hexokinase 1, Hexokinase type I, HK I, Hexokinase-A) is encoded by the HK1 gene (Gene ID: 3098) in human. Hexokinase I (HKI) belongs to the hexokinase family that has four isoforms. HKI and II exhibit overlapping tissue expression, but different subcellular distribution. HKI is a monomeric enzyme that associates with outer mitochondrial membrane and its mitochondrial binding peptide is localized in amino acids 1-10. HKII is associated with both mitochondrial and cytoplasmic compartments. Hexokinase catalyzes the phosphorylation of glucose and other hexoses to produce glucose-6-phosphate (G-6-P) and other hexose phosphates. Both HKI and II are inhibited allosterically by G-6-P and this sensitivity is reported to decrease when HK are bound to mitochondria. The N- and C-terminal halves of HK contains a hexokinase domain (aa 16-458 and 464-906). Its catalytic activity is associated with the C-terminus while regulatory function is associated with the N-terminus half. Each domain can bind a single D-glucose and D-glucose 6-phosphate molecule. Hexokinase exhibits lower apparent Km values for ATP generated in the mitochondria than for ATP provided exogenously indicating that the active site of bound hexokinase is relatively inaccessible to extramitochondrial ATP. Hence, mitochondrial bound hexokinase can sequentially accept mitochondrially generated ATP in a kinetically advantageous way. This ZooMAb^® recombinant monoclonal antibody, generated by our propriety technology, offers significantly enhanced specificity, Affinity^™, reproducibility, and stability over conventional monoclonals. (Ref.: John, S., et al. (2011). PLoS One. 6(3); e17674; Aleshin, AE., et al. (1998). Structure. 6(1); 39-50; Gots, RE., and Bessman, SP^®. (1974). Arch. Biochem. Biophys. 163(1); 7-14).

Physical form

Purified recombinant rabbit monoclonal antibody IgG, lyophilized in PBS, 5% Trehalose, normal appearance a coarse or translucent resin. The PBS/trehalose components in the ZooMAb formulation can have the appearance of a semi-solid (bead like gel) after lyophilization. This is a normal phenomenon. Please follow the recommended reconstitution procedure in the data sheet to dissolve the semi-solid, bead-like, gel-appearing material. The resulting antibody solution is completely stable and functional as proven by full functional testing. Contains no biocide or preservatives, such as azide, or any animal by-products. Larger pack sizes provided as multiples of 25 µL.

Reconstitution

300 µg/mL after reconstitution at 25 µL per vial. Please refer to guidance on suggested starting dilutions and/or titers per application and sample type.

Storage and Stability

Recommend storage of lyophilized product at 2-8°C; Before reconstitution, micro-centrifuge vials briefly to spin down material to bottom of the vial; Reconstitute each vial by adding 25 µL of filtered lab grade water or PBS; Reconstituted antibodies can be stored at 2-8°C, or -20°C for long term storage. Avoid repeated freeze-thaws.

Other Notes

Concentration: Please refer to the Certificate of Analysis for the lot-specific concentration.

Legal Information

Affinity is a trademark of Mine Safety Appliances Co.

SP is a registered trademark of Sigma-Aldrich Co. LLC

ZooMAb is a registered trademark of Merck KGaA, Darmstadt, Germany

Disclaimer

Unless otherwise stated in our catalog or other company documentation accompanying the product(s), our products are intended for research use only and are not to be used for any other purpose, which includes but is not limited to, unauthorized commercial uses, in vitro diagnostic uses, ex vivo or in vivo therapeutic uses or any type of consumption or application to humans or animals.