Human EEA1 knockout HeLa cell line
- Advanced Validation
- 각 태그의 뜻
Be the first to review this product! 리뷰 제출
|
(0 출판물)
- WB
Lab
Western blot - Human EEA1 knockout HeLa cell line (AB261822)
Lanes 1- 4 : Merged signal (red and green). Green - ab109110 observed at 175 kDa. Red - Anti-GAPDH antibody [6C5] - Loading Control (ab8245) observed at 37 kDa.
ab109110 was shown to react with EEA1 in wild-type HeLa cells in western blot. The band observed in CRISPR/Cas9 edited cell line ab261822 (CRISPR/Cas9 edited cell lysate ab256897) lane below 175kDa may represent truncated forms and cleaved fragments. This has not been investigated further. Wild-type HeLa and EEA1 CRISPR/Cas9 edited HeLa cell lysates were subjected to SDS-PAGE. Membrane was blocked for 1 hour at room temperature in 0.1% TBST with 3% non-fat dried milk. ab109110 and Anti-GAPDH antibody [6C5] - Loading Control (ab8245) were incubated overnight at 4° at a 1 in 10000 dilution and a 1 in 20000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye®800CW) preadsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye®680RD) preadsorbed (ab216776) secondary antibodies at 1 in 20000 dilution for 1 hour at room temperature before imaging.
All lanes:
Western blot - Anti-EEA1 antibody [EPR4245] - Early Endosome Marker (<a href='/ko/products/primary-antibodies/eea1-antibody-epr4245-early-endosome-marker-ab109110'>ab109110</a>) at 1/1000 dilution
Lane 1:
Wild-type HeLa cell lysate at 20 µg
Lane 2:
EEA1 CRISPR/Cas9 edited HeLa cell lysate at 20 µg
Lane 2:
Western blot - Human EEA1 knockout HeLa cell line (ab261822)
Lane 3:
Daudi cell lysate at 20 µg
Lane 4:
SH-SY5Y cell lysate at 20 µg
Predicted band size: 162 kDa
Observed band size: 175 kDa
false
- Sanger seq
Unknown
Sanger Sequencing - Human EEA1 knockout HeLa cell line (AB261822)
Homozygous : 1 bp insertion in exon 13.
보완 제품
Cell Lines & Lysates
AB261794
Human HIST1H1C (Histone H1.2) knockout HeLa cell line
Advanced Validation
- 0
- 0
Cell Lines & Lysates
AB261873
Human HIST1H1C (Histone H1.2) knockout A549 cell line
Advanced Validation
- 0
- 0
Reactivity 정보
제품 세부 정보
We will provide viable cells that proliferate on revival.
Western blot data indicates that the CRISPR gene edit may have resulted in a truncation of the protein of interest. Please see data images.
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.
제품 구성
특성 및 보관 정보
유전자명
유전자 편집 종류
유전자 편집 방식
Knockout 검증
Zygosity
품질 관리 - STR analysis
배송 시 보관 조건
적절한 단기 보관 조건
적절한 장기 보관 조건
추가 정보
This supplementary information is collated from multiple sources and compiled automatically.
Biological function summary
EEA1 plays a significant role in early endosomal pathways facilitating endocytic vesicle fusion. EEA1 does not usually work alone; it functions as part of a larger complex involving other endosomal proteins. This protein acts as a tethering molecule which stabilizes the interaction of vesicles with endosomes. By anchoring vesicles EEA1 ensures the correct delivery and sorting of cargo within the cell which is important for cellular homeostasis and signaling.
Pathways
EEA1 contributes to the endocytic pathway and receptor recycling. It ensures proper functioning of the endocytic recycling of receptors such as the epidermal growth factor receptor (EGFR). EEA1-mediated tethering is vital for these pathways and it works closely with proteins like Rab5 which regulate endocytic trafficking. Through these interactions EEA1 affects cellular processes by controlling the flow of signaling receptors and other molecules within the cell.
Cell culture
Biosafety level
EU: 2 US: 2
Adherent/suspension
Adherent
Gender
Female
초기 취급 지침
Upon arrival, the vial should be stored in liquid nitrogen vapor phase and not at -80°C. Storage at -80°C may result in loss of viability. <p>1. Thaw the vial in 37°C water bath for approximately 1-2 minutes.<br>2. Transfer the cell suspension (0.8 mL) to a 15 mL/50 mL conical sterile polypropylene centrifuge tube containing 8.4 mL pre-warmed culture medium, wash vial with an additional 0.8 mL culture medium (total volume 10 mL) to collect remaining cells, and centrifuge at 201 x g (rcf) for 5 minutes at room temperature. 10 mL represents minimum recommended dilution. 20 mL represents maximum recommended dilution.<br>3. Resuspend the cell pellet in 5 mL pre-warmed culture medium and count using a haemocytometer or alternative cell counting method seed all remaining cells into a T25.<br>4. Incubate the culture at 37°C incubator with 5% CO<sub>2</sub>. Check the culture one day after revival and continue to check until 80% confluent. Media change can be given if needed.<br>5. Once confluent passage into an appropriate flask at a density of 2x10<sup>4</sup> cells/cm<sup>2</sup>. Seeding density is given as a guide only and should be scaled to align with individual lab schedules. Cultures should be monitored daily.</p>
Subculture 가이드라인
- All seeding densities should be based on cell counts gained by established methods.
- A guide seeding density of 2x104 cells/cm2 is recommended.
- Cells should be passaged when they have achieved 80-90% confluence.
배양 배지
DMEM (High Glucose) + 10% FBS
동결보존 배지
Cell Freezing Medium-DMSO Serum free media, contains 8.7% DMSO in MEM supplemented with methyl cellulose.
타겟 정보
대체 명칭 보기
Product promise
Please note: All products are 'FOR RESEARCH USE ONLY. NOT FOR USE IN DIAGNOSTIC OR THERAPEUTIC PROCEDURES'.
For licensing inquiries, please contact partnerships@abcam.com