人CBFB knockout A-431 cell裂解物(ab270495)
概述
-
产品名称
人CBFB knockout A-431 cell裂解物 -
产品概述
Knockout cell lysate achieved by CRISPR/Cas9.
-
Parental Cell Line
A431 -
Organism
Human -
Mutation description
Knockout achieved by CRISPR/Cas9; X = 19 bp deletion; Frameshift = 99.99% -
Passage number
<20 -
Knockout validation
Next Generation Sequencing (NGS), 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.
-
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 ab280548 - Human CBFB knockout A-431 cell lysate 1 x 100µg ab263973 - Human wild-type A-431 cell lysate 1 x 100µg -
研究领域
-
Cell type
epithelial -
Disease
Epidermoid Carcinoma -
Gender
Female
靶标
-
功能
CBF binds to the core site, 5'-PYGPYGGT-3', of a number of enhancers and promoters, including murine leukemia virus, polyomavirus enhancer, T-cell receptor enhancers, LCK, IL3 and GM-CSF promoters. CBFB enhances DNA binding by RUNX1. -
疾病相关
Note=A chromosomal aberration involving CBFB is associated with acute myeloid leukemia of M4EO subtype. Pericentric inversion inv(16)(p13;q22). The inversion produces a fusion protein that consists of the 165 N-terminal residues of CBF-beta (PEPB2) with the tail region of MYH11. -
序列相似性
Belongs to the CBF-beta family. -
细胞定位
Nucleus. - Information by UniProt
-
别名
- CBF b
- CBF beta
- CBF-beta
see all
相关产品
-
KO cell lines
-
Related Products
应用
The Abpromise guarantee
Abpromise™承诺保证使用ab270495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.
|
说明 |
---|
WB
Use at an assay dependent concentration. |
图片
-
Knockout achieved by CRISPR/Cas9; X = 19 bp deletion; Frameshift = 99.99%
-
Lane 1: Wild-type A431 cell lysate 20 ug
Lane 2: CBFB knockout A431 cell lysate 20 ug
Lane 3: K562 (Human chronic myelogenous leukemia lymphoblast cell line ) whole cell lysate 20 ug
Lane 4: HeLa (Human epithelial cell line from cervix adenocarcinoma) whole cell lysate 20 ug
ab133600 was shown to react with CBFb in wild-type A-431 cells in western blot. Loss of signal was observed when CBFB knockout cell line ab270472 (knockout cell lysate ab270495) was used. Membranes were blocked in 2 % BSA in TBS-T (0.1 % Tween®) before incubation with ab133600 overnight at 4°C at a 1 in 1000 dilution and ab184095 (Mouse Anti-GAPDH antibody [mAbcam 9484] - Alexa Fluor® 680) at a 1 in 1000 dilution. Blots were incubated with HRP conjugated Goat anti-Rabbit (H+L) secondary antibody at 1/5000 for 1 hour at room temperature before development with Optiblot ECL reagent (ab133456) and imaging. -
Lane 1: Wild-type A431 cell lysate 20 ug
Lane 2: CBFB knockout A431 cell lysate 20 ug
Lane 3: K562 (Human chronic myelogenous leukemia lymphoblast cell line ) whole cell lysate 20 ug
Lane 4: HeLa (Human epithelial cell line from cervix adenocarcinoma) whole cell lysate 20 ug
Lanes 1 - 4: Merged signal (red and green). Green - ab133600 observed at 22 kDa. Red - loading control ab7291 (Mouse anti-Alpha Tubulin [DM1A]) observed at 55kDa.
ab133600 was shown to react with CBFb in wild-type A-431 cells in western blot with loss of signal observed in CBFB knockout cell line ab270472 (knockout cell lysate ab270495). Wild-type and CBFB knockout A-431 cell lysates were subjected to SDS-PAGE. Membranes were blocked in 2% BSA in TBS-T (0.1% Tween®) before incubation with ab133600 and ab7291 (Mouse anti-Alpha Tubulin [DM1A]) 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. -
Lane 1: Wild-type A431 cell lysate 20 ug
Lane 2: CBFB knockout A431 cell lysate 20 ug
Lane 3: K562 (Human chronic myelogenous leukemia lymphoblast cell line ) whole cell lysate 20 ug
Lane 4: HeLa (Human epithelial cell line from cervix adenocarcinoma) whole cell lysate 20 ug
Lanes 1 - 4: Merged signal (red and green). Green - ab124693 observed at 24-25 kDa. Red - loading control, ab7291 (Mouse anti-Alpha Tubulin [DM1A] observed at 55kDa.
ab124693 was shown to react with CBFb in wild-type A-431 cells in western blot Loss of signal was observed when CBFB knockout cell line ab270472 (knockout cell lysate ab270495) was used. Wild-type and CBFB knockout cell lysates were subjected to SDS-PAGE. Membranes were blocked in 3% milk in TBS-T (0.1% Tween®) before incubation with ab124693 and ab7291 (Mouse anti-Alpha Tubulin [DM1A] 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.
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)
ab270495 尚未被引用在任何文献中。