Invitrogen

HK1 Recombinant Polyclonal Antibody

Advanced Verification

This Antibody was verified by Orthogonal method to ensure that the antibody binds to the antigen stated. View Details

View all (18) HK1 antibodies

Datasheet

Questions & Answers

Datasheet

Questions & Answers

Product Image Gallery

HK1 Antibody in Western Blot (WB) — Western blot analysis was performed on Whole cell extracts (60 µg lysate) of PC-3 (Lane 1), K-562 (Lane 2), U-87 MG (Lane 3), MCF-7 (Lane 4), A-431 (Lane 5) and A549 (Lane 6). The blots were probed with Anti-Hexokinase 1 Recombinant Rabbit Polyclonal Antibody (Product # 711777, 2.5 µg/mL) and detected by chemiluminescence using Goat anti-Rabbit IgG (H+L) Superclonal™ Secondary Antibody, HRP conjugate (Product # A27036, 0.25 µg/mL, 1:4000 dilution). A 102 kDa band corresponding to Hexokinase 1 was observed across the cell lines tested. Known quantity of protein samples were electrophoresed using Novex®NuPAGE®4-12% Bis-Tris gel (Product # NP0322BOX), 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 iBlot® Dry Blotting System (Product # IB21001). 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).

HK1 Antibody in Immunofluorescence (IF) — For immunofluorescence analysis, MCF7 cells were fixed and permeabilized for detection of endogenous Hexokinase-1 using Anti-Hexokinase-1 Recombinant Rabbit Polyclonal Antibody (Product # 711777, 5 µg/mL) and labeled with Goat anti-Rabbit IgG (H+L) Superclonal™ Secondary Antibody, Alexa Fluor® 488 conjugate (Product # A27034, 1:2000). Panel a) shows representative cells that were stained for detection and localization of Hexokinase-1 protein (green), Panel b) is stained for nuclei (blue) using SlowFade® Gold Antifade Mountant with DAPI (Product # S36938). Panel c) represents mitochondrial staining using MitoTracker® Red CMXRos (Product # M7512). Panel d) is a composite image of Panels a, b and c clearly demonstrating co-localization of Hexokinase-1 with mitotracker which specifically binds to the mitochondria. Panel e) represents control cells without primary antibody to assess background. The images were captured at 60X magnification.

HK1 Antibody in Orthogonal method — Antibody specificity was demonstrated by showing similar results, obtained through an antibody dependent and an antibody independent method. Immunofluorescence analysis using Anti-Hexokinase 1 Recombinant Rabbit Polyclonal Antibody (Product # 711777, 5 µg/mL) shows co- localization with a mitochondrial marker, MitoTracker® Red CMXRos (Product # M7512) (antibody independent method) in MCF-7 cells. {OM}

Product Details

Applications

Tested Dilution

Publications

Western Blot (WB)

2.5 µg/mL

Immunocytochemistry (ICC)

5 µg/mL

Immunofluorescence (IF)

5 µg/mL

Product Specifications

Species Reactivity

Human

Host/Isotype

Rabbit / IgG

Class

Recombinant Polyclonal

Type

Antibody

Immunogen

Peptides corresponding to human HK1 [1) aa315-aa334 2) aa719-aa738]

Conjugate

Unconjugated

Form

Liquid

Concentration

0.5 mg/mL

Purification

Protein A

Storage buffer

PBS, pH 7.4

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_2724453

Product Specific Information

This antibody is predicted to react with Monkey, Horse, Dog, Bovine.

Recombinant rabbit polyclonal antibodies are unique offerings from Thermo Fisher Scientific. They are comprised of a selection of multiple different recombinant monoclonal antibodies, providing the best of both worlds - the sensitivity of polyclonal antibodies with the specificity of monoclonal antibodies - all delivered with the consistency only found in a recombinant antibody. While functionally the same as a polyclonal antibody - recognizing multiple epitope sites on the target and producing higher detection sensitivity for low abundance targets - a recombinant rabbit polyclonal antibody has a known mixture of light and heavy chains. The exact population can be produced in every lot, circumventing the biological variability typically associated with polyclonal antibody production.

Target Information

This gene encodes a member of the ARHGAP family of Rho/Rac/Cdc42-like GTPase activating proteins. The encoded protein interacts with the Ras-related protein Gem through its N-terminal domain. Separately, it interacts with RhoA through a RhoGAP domain, and stimulates RhoA-dependent GTPase activity. Alternatively spliced variants that encode different protein isoforms have been described but their full-length nature has not been determined.

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