人HK2 (Hexokinase II) knockout HEK-293 cell裂解物(ab281353)
概述
-
产品名称
人HK2 (Hexokinase II) knockout HEK-293 cell裂解物
参阅全部 Hexokinase II 试剂盒 -
产品概述
Western blot data indicates that the CRISPR gene edit may have resulted in a truncation of the protein of interest. Please see data images.
-
Parental Cell Line
HEK-293 -
Organism
Human -
Mutation description
Knockout achieved by CRISPR/Cas9; X = 5 bp deletion; Frameshift: 99% -
Passage number
<20 -
Knockout validation
Next Generation Sequencing (NGS) -
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.
-
说明
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 and ERS Genomics Limited, and is developed with patented technology. For full details of the limited use licenses and relevant patents please refer to our limited use license and patent pages.
-
经测试应用
适用于: WBmore details
性能
-
存放说明
Store at -80°C. Please refer to protocols. -
组件 1 kit ab281393 - Human HK2 knockout HEK-293 cell lysate 1 x 100µg ab259780 - Human wild-type HEK-293 cell lysate 1 x 100µg -
研究领域
-
Cell type
epithelial -
Gender
Female
靶标
-
组织特异性
Predominant hexokinase isozyme expressed in insulin-responsive tissues such as skeletal muscle. -
通路
Carbohydrate metabolism; hexose metabolism. -
序列相似性
Belongs to the hexokinase family.
Contains 2 hexokinase domains. -
结构域
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. -
细胞定位
Mitochondrion outer membrane. Its hydrophobic N-terminal sequence may be involved in membrane binding. - Information by UniProt
-
别名
- DKFZp686M1669
- Hexokinase 2
- Hexokinase 2 muscle
see all
相关产品
-
KO cell lines
-
Related Products
应用
The Abpromise guarantee
Abpromise™承诺保证使用ab281353于以下的经测试应用
“应用说明”部分 下显示的仅为推荐的起始稀释度；实际最佳的稀释度/浓度应由使用者检定。
| 应用 | Ab评论 | 说明 |
|---|---|---|
| WB |
Use at an assay dependent concentration.
Western blot data indicates that the CRISPR gene edit may have resulted in a truncation of the protein of interest. Please see data images. |
| 说明 |
|---|
|
WB
Use at an assay dependent concentration. Western blot data indicates that the CRISPR gene edit may have resulted in a truncation of the protein of interest. Please see data images. |
图片
-
Western blot - Human HK2 (Hexokinase II) knockout HEK-293 cell lysate (ab281353)
Lane 1: Wild-type HEK-293 (Human epithelial cell line from embryonic kidney) whole cell lysate 20 ug
Lane 2: HK2 knockout HEK-293 (Human epithelial cell line from embryonic kidney) whole cell lysate 20 ug
Lane 3: HeLa (Human epithelial cell line from cervix adenocarcinoma) whole cell lysate 20 ug
Lane 4: A549 (Human lung carcinoma cell line) whole cell lysate 20 ug
Lanes 1 - 4: Merged signal (red and green). Green - ab209847 observed at 110 kDa. Red - loading control ab8245 (Mouse anti-GAPDH antibody [6C5]) observed at 37kDa.
ab209847 was shown to bind specifically to HK2. A band was observed at 110 kDa in wild-type cell lysates with no signal at this size in HK2 knockout cell line ab269485 (knockout cell lysate ab281353). Wild-type and HK2 knockout HEK-293 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. -
Next Generation Sequencing - Human HK2 (Hexokinase II) knockout HEK-293 cell lysate (ab281353)Knockout achieved by CRISPR/Cas9; X = 5 bp deletion; Frameshift: 99%
实验方案
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.
数据表及文件
-
SDS download
-
Datasheet download
文献 (0)
ab281353 尚未被引用在任何文献中。