Invitrogen

HK2 Recombinant Rabbit Monoclonal Antibody (2H8L6)

Advanced Verification

This Antibody was verified by Knockdown to ensure that the antibody binds to the antigen stated.

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Datasheet

Protocols

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Datasheet

Protocols

Questions & Answers

Tech Support

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HK2 Antibody in Immunocytochemistry (ICC/IF) — Immunofluorescence analysis of Hexokinase-2 was performed using 70% confluent log phase A-431 cells. The cells were fixed with 4% paraformaldehyde for 10 minutes, permeabilized with 0.01% Triton™ X-100 for 15 minutes, and blocked with 2% BSA for 45 minutes at room temperature. The cells were labeled with HK2 Recombinant Rabbit Monoclonal Antibody (2H8L6) (Product # 700422) at 1:100 in 0.1% BSA, incubated at 4 degree celsius overnight and then labeled with Donkey anti-Rabbit IgG (H+L) Highly Cross-Adsorbed Secondary Antibody, Alexa Fluor Plus 488 (Product # A32790), (1:2000), for 45 minutes at room temperature (Panel a: Green). Nuclei (Panel b:Blue) were stained with Hoechst 33342 (Product # H1399). F-actin (Panel c: Red) was stained with Rhodamine Phalloidin (Product # R415, 1:300). Panel d represents the merged image showing cytoplasmic localization. Panel e represents control cells with no primary antibody to assess background. The images were captured at 40X magnification in CellInsight CX7 LZR High-Content Screening (HCS) Platform (Product # CX7A1110LZR) and externally deconvoluted (D.Sage et al. / Methods 115 (2017) 28-41).

HK2 Antibody in Western Blot (WB) — Western blot analysis was performed on whole cell extracts (30 µg lysate) of HCT 116 (Lane 1), HT -29 (lane 2), SK-O-V3 (lane 3), T47D (Lane 4), MDA-MB-231 (lane 5), and MCF-7 (Lane 6).The blots were probed with Anti-Hexokinase II Rabbit Monoclonal Antibody (Product # 700422, 3 µg/mL) and detected by chemiluminescence Goat Anti-Rabbit IgG Secondary Antibody, HRP conjugate (Product # G-21234, 1:5000 dilution). A 102kDa band corresponding to Hexokinase II was observed across cell lines tested. Known quantity of protein samples were electrophoresed using Novex® NuPAGE® 10 % Bis-Tris gel (Product # NP0301BOX), XCell SureLock™ Electrophoresis System (Product # EI0002) and Novex® Sharp Pre-Stained Protein Standard (Product # LC5800). Resolved proteins were then transferred onto a nitrocellulose membrane with Pierce™ Power Blotter System (Product # 22834). The membrane was probed with the relevant primary and secondary Antibody following blocking with 5 % skimmed milk. Chemiluminescent detection was performed using Pierce™ ECL Western Blotting Substrate (Product # 32106).

HK2 Antibody — Antibody specificity was demonstrated by siRNA mediated knockdown of target protein. U-87 MG cells were transfected with Hexokinase-2 siRNA and decrease in signal intensity was observed in Western Blot application using Anti-HK2 Recombinant Rabbit Monoclonal Antibody (2H8L6) (Product # 700422). {KD}

Product Details

700422

Applications	Tested Dilution	Publications
Western Blot (WB)	2-3 µg/mL	-
Immunocytochemistry (ICC/IF)	2-3 µg/mL	-

Product Specifications
Species Reactivity	Human
Host/Isotype	Rabbit / IgG
Class	Recombinant Monoclonal
Type	Antibody
Clone	2H8L6
Immunogen	A recombinant protein corresponding to amino acids 325-444. View antigen
Conjugate	Unconjugated
Form	Liquid
Concentration	0.5 mg/mL
Purification	Protein A
Storage buffer	PBS
Contains	0.09% sodium azide
Storage conditions	Store at 4°C short term. For long term storage, store at -20°C, avoiding freeze/thaw cycles.
RRID	AB_2532323

Product Specific Information

This antibody is predicted to react with bovine, mouse and rat based on sequence homology.

Intact IgG appears on a non-reducing gel as ~150 kDa band and upon reduction generating a ~25 kDa light chain band and a ~50 kDa heavy chain.

Recombinant rabbit monoclonal antibodies are produced using in vitro expression systems. The expression systems are developed by cloning in the specific antibody DNA sequences from immunoreactive rabbits. Then, individual clones are screened to select the best candidates for production. The advantages of using recombinant rabbit monoclonal antibodies include: better specificity and sensitivity, lot-to-lot consistency, animal origin-free formulations, and broader immunoreactivity to diverse targets due to larger rabbit immune repertoire.

Target Information

In vertebrates there are four major glucose-phosphorylating isoenzymes, designated hexokinase I, II, III, and IV. Hexokinase activity is involved in the first step in several metabolic pathways including phosphorylation of glucose to produce glucose-6-phosphate, thus committing glucose to the glycolytic pathway. Hexokinase is an allosteric enzyme inhibited by its product GLC-6-P. Hexokinase 2 is the predominant hexokinase isozyme expressed in insulin-responsive tissues such as skeletal muscle. Expression of this gene is insulin-responsive, and studies in rat suggest that it is involved in the increased rate of glycolysis seen in rapidly growing cancer cells.

For Research Use Only. Not for use in diagnostic procedures. Not for resale without express authorization.

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Bioinformatics

Protein Aliases: DKFZp686M1669; Hexokinase; Hexokinase type II; Hexokinase-2; hexokinase-2 muscle; hexokinase-2, muscle; Hexokinase-B; HK II; Muscle form hexokinase; OTTHUMP00000202738

Gene Aliases: HK2; HKII; HXK2

UniProt ID: (Human) P52789

Entrez Gene ID: (Human) 3099

Molecular Function: kinase