Human TPBG knockout MCF7 cell lysate (ab269661)
製品の概要
-
製品名
Human TPBG knockout MCF7 cell lysate -
製品の概要
Knockout cell lysate achieved by CRISPR/Cas9.
-
Parental Cell Line
MCF7 -
Organism
Human -
Mutation description
Knockout achieved by CRISPR/Cas9; X = 1 bp insertion; Frameshift: 97% -
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.
-
特記事項
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.
-
アプリケーション
適用あり: WBmore details
製品の特性
-
保存方法
Store at -80°C. Please refer to protocols. -
内容 1 kit ab280536 - Human TPBG knockout MCF7 cell lysate 1 x 100µg ab269599 - Human wild-type MCF7 cell lysate 1 x 100µg -
研究分野
-
Cell type
epithelial -
Disease
Adenocarcinoma -
Gender
Female
ターゲット情報
-
組織特異性
Expressed by all types of trophoblasts as early as 9 weeks of development. Specific for trophoblastic cells except for amniotic epithelium. In adult tissues, the expression is limited to a few epithelial cell types but is found on a variety of carcinoma. -
配列類似性
Contains 6 LRR (leucine-rich) repeats.
Contains 1 LRRCT domain.
Contains 1 LRRNT domain. -
細胞内局在
Membrane. - Information by UniProt
-
製品の状態
5T4 is a 72 kDa transmembrane glycoprotein that exhibits restricted expression in human and mouse adult tissues, is upregulated on many carcinomas and tumour expression correlates with poorer clinical outcome in ovarian, gastric and colorectal cancers. mES cells lack cell surface expression of the 5T4 antigen, and both protein and mRNA are rapidly upregulated following induction of differentiation. This proteins expression correlates with downregulation of OCT-4 and Tra-1–60 (Ward et al., 2006). -
別名
- 5T4
- 5T4 oncofetal antigen
- 5T4 oncofetal trophoblast glycoprotein
see all
関連製品
-
KO cell lines
-
Related Products
アプリケーション
The Abpromise guarantee
Abpromise保証は、 次のテスト済みアプリケーションにおけるab269661の使用に適用されます
アプリケーションノートには、推奨の開始希釈率がありますが、適切な希釈率につきましてはご検討ください。
アプリケーション | Abreviews | 特記事項 |
---|---|---|
WB |
Use at an assay dependent concentration. Predicted molecular weight: 46 kDa.
|
特記事項 |
---|
WB
Use at an assay dependent concentration. Predicted molecular weight: 46 kDa. |
画像
-
Knockout achieved by CRISPR/Cas9; X = 1 bp insertion; Frameshift: 97%
-
Lane 1: Wild-type MCF7 (Human breast adenocarcinoma cell line) whole cell lysate, 20 ug
Lane 2: TPBG knockout MCF7 (Human breast adenocarcinoma cell line) whole cell lysate, 20 ug
Lane 3: HeLa (Human epithelial cell line from cervix adenocarcinoma) whole cell lysate, 20 ug
Lane 4: HEK-293T (Human epithelial cell line from embryonic kidney transformed with large T antigen) whole cell lysate, 20 ugab199547 was shown to react with 5T4 (HRP) in wild-type MCF7 cells in western blot. Loss of signal was observed when TPBG knockout cell line ab269499 (knockout cell lysate ab269661) was used. Membranes were blocked in 3 % milk in TBS-T (0.1 % Tween®) before incubation with ab199547 overnight at 4 °C at a 1 in 5000 dilution Blots were developed with Optiblot ECL reagent (ab133456) and imaged.
-
Lane 1: Wild-type MCF7 (Human breast adenocarcinoma cell line) whole cell lysate, 20 ug
Lane 2: TPBG knockout MCF7 (Human breast adenocarcinoma cell line) whole cell lysate, 20 ug
Lane 3: HeLa (Human epithelial cell line from cervix adenocarcinoma) whole cell lysate, 20 ug
Lane 4: HEK-293T (Human epithelial cell line from embryonic kidney transformed with large T antigen) whole cell lysate, 20 ugLanes 1 - 4: Merged signal (red and green). Green - ab134162 observed at 85 kDa. Red - loading control ab8245 (Mouse anti-GAPDH antibody [6C5]) observed at 37 kDa.
ab134162 was shown to react with 5T4 in wild-type MCF7 cells in Western blot with loss of signal observed in TPBG knockout cell line ab269499 (knockout cell lysate ab269661). Wild-type MCF7 and TPBG knockout cell lysates were subjected to SDS-PAGE. Membranes were blocked in before incubation with ab134162 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 h at room temperature before imaging.
-
Lane 1: Wild-type MCF7 (Human breast adenocarcinoma cell line) whole cell lysate, 20 ug
Lane 2: TPBG knockout MCF7 (Human breast adenocarcinoma cell line) whole cell lysate, 20 ug
Lane 3: HeLa (Human epithelial cell line from cervix adenocarcinoma) whole cell lysate, 20 ug
Lane 4: HEK-293T (Human epithelial cell line from embryonic kidney transformed with large T antigen) whole cell lysate, 20 ugLanes 1 - 4: Merged signal (red and green). Green - ab129058 observed at 85 kDa. Red - loading control ab8245 (Mouse anti-GAPDH antibody [6C5]) observed at 37 kDa.
ab129058 was shown to react with 5T4 in wild-type MCF7 cells in Western blot with loss of signal observed in TPBG knockout cell line ab269499 (knockout cell lysate ab269661). Wild-type MCF7 and TPBG knockout cell lysates were subjected to SDS-PAGE. Membranes were blocked in before incubation with ab129058 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 h at room temperature before imaging.
プロトコール
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.
データシートおよび資料
-
SDS download
-
Datasheet download
参考文献 (0)
ab269661 は論文での使用が確認できていません。