Human HK2 (Hexokinase II) knockout HCT116 cell lysate (ab275250)
Overview
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Product name
Human HK2 (Hexokinase II) knockout HCT116 cell lysate
See all Hexokinase II kits -
Product overview
Knockout cell lysate achieved by CRISPR/Cas9. -
Parental Cell Line
HCT116 -
Organism
Human -
Mutation description
Knockout achieved by using CRISPR/Cas9, Homozygous: 1 bp insertion in exon 2 -
Passage number
<20 -
Knockout validation
Sanger Sequencing, Western Blot (WB) -
Reconstitution notes
To use as WB control, resuspend the lyophilizate in 50 µL of LDS* Sample Buffer to have a final concentration of 2 mg/ml. For reducing conditions, we recommend a final concentration of 0.1 M DTT.*Usage of SDS sample buffer is not recommended with these lyophilized lysates.
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Notes
Lysate preparation: Our lysates are made using RIPA buffer to which we add a protease inhibitor cocktail and phosphatase inhibitor cocktail (ratio: 300:100:10). This means that the protein of interest is denatured. If you require a native form of the protein please use the live cell version - found here. Please refer to our lysis protocol for further details on how our lysates are prepared.
User storage instructions: Lyophilizate may be stored at 4°C. After reconstitution, store at -20°C for short-term storage or -80°C for long-term storage.
Access thousands of knockout cell lysates, generated from commonly used cancer cell lines.
See here for more information on knockout cell lysates.Abcam has not and does not intend to apply for the REACH Authorisation of customers’ uses of products that contain European Authorisation list (Annex XIV) substances.
It is the responsibility of our customers to check the necessity of application of REACH Authorisation, and any other relevant authorisations, for their intended uses.This product is subject to limited use licenses from The Broad Institute, ERS Genomics Limited and Sigma-Aldrich Co. LLC, and is developed with patented technology. For full details of the licenses and patents please refer to our limited use license and patent pages.
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Tested applications
Suitable for: WBmore details
Properties
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Storage instructions
Store at -80°C. Please refer to protocols. -
Components 1 kit ab277294 - Human HK2 knockout HCT116 cell lysate 1 x 100µg ab277289 - Human wild-type HCT116 cell lysate 1 x 100µg -
Research areas
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Cell type
epithelial -
Disease
Carcinoma -
Gender
Male
Target
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Tissue specificity
Predominant hexokinase isozyme expressed in insulin-responsive tissues such as skeletal muscle. -
Pathway
Carbohydrate metabolism; hexose metabolism. -
Sequence similarities
Belongs to the hexokinase family.
Contains 2 hexokinase domains. -
Domain
The N- and C-terminal halves of this hexokinase show extensive sequence similarity to each other. The catalytic activity is associated with the C-terminus while regulatory function is associated with the N-terminus. Each domain can bind a single glucose and Gluc-6-P molecule. -
Cellular localization
Mitochondrion outer membrane. Its hydrophobic N-terminal sequence may be involved in membrane binding. - Information by UniProt
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Alternative names
- DKFZp686M1669
- Hexokinase 2
- Hexokinase 2 muscle
see all
Associated products
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KO cell lines
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Related Products
Applications
The Abpromise guarantee
Our Abpromise guarantee covers the use of ab275250 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
| Application | Abreviews | Notes |
|---|---|---|
| WB |
Use at an assay dependent concentration. Predicted molecular weight: 102 kDa.
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| Notes |
|---|
|
WB
Use at an assay dependent concentration. Predicted molecular weight: 102 kDa. |
Images
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Western blot - Human HK2 (Hexokinase II) knockout HCT116 cell lysate (ab275250)
Lane 1: Wild-type HCT116 cell lysate 20 ug
Lane 2: HK2 knockout HCT116 cell lysate 20 ug
Lane 3: HeLa cell lysate 20 ug
Lane 4: SH-SY5Y cell lysate 20 ug
Lanes 1 - 4:Merged signal (red and green). Green - ab209847 observed at 105 kDa. Red - loading control, ab8245 (Mouse anti-GAPDH antibody [6C5]) observed at 37kDa.
ab209847 was shown to react with Anti-Hexokinase II in wild-type HCT 116 cells in western blot with loss of signal observed in HK2 knockout cell line ab273721 (HK2 knockout cell lysate ab275250). HCT 116 wild-type and HK2 knockout cell lysates were subjected to SDS-PAGE. Membranes were blocked in 3% milk in TBS-T (0.1% Tween®) before incubation with ab209847 and ab8245 (Mouse anti-GAPDH antibody [6C5]) overnight at 4°C at a 1 in 1000 dilution and a 1 in 20000 dilution respectively. Blots were incubated with Goat anti-Rabbit IgG H&L (IRDye® 800CW) preabsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye® 680RD) preabsorbed (ab216776) secondary antibodies at 1 in 20000 dilution for 1 hour at room temperature before imaging. -
Sanger Sequencing - Human HK2 (Hexokinase II) knockout HCT116 cell lysate (ab275250)Allele-1: 1 bp insertion in exon 2.
Protocols
To our knowledge, customised protocols are not required for this product. Please try the standard protocols listed below and let us know how you get on.
Datasheets and documents
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SDS download
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Datasheet download
References (0)
ab275250 has not yet been referenced specifically in any publications.