Human EXOC2 knockout HeLa cell line
Human EXOC2 knockout HeLa cell line
- Advanced Validation
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EXOC2 KO cell line available to order. KO validated. Free of charge wild type control available. Knockout achieved by using CRISPR/Cas9, 1 bp insertion in exon 3 and 2 bp insertion in exon 3. To order both knockout and wild-type control cells: select '2 x 1000000 Cells/vial'. To order only knockout cells: select '1000000 Cells/vial'.
別名を表示する
EXOC2_HUMAN, Exocyst complex component 2, Exocyst complex component Sec5, FLJ11026, SEC5, SEC5 like 1, SEC5L1, Sec5p
- WB
Lab
Western blot - Human EXOC2 knockout HeLa cell line (AB265839)
Lanes 1- 4 : Merged signal (red and green). Green - ab140620 observed at 104 kDa. Red - Anti-GAPDH antibody [6C5] - Loading Control (ab8245) observed at 37 kDa.
ab140620 was shown to react with EXOC2 in wild-type HeLa cells in western blot. The band observed in CRISPR/Cas9 edited cell line ab265839 (CRISPR/Cas9 edited cell lysate ab257944) lane below 104kDa may represent truncated forms and cleaved fragments. This has not been investigated further. Wild-type HeLa and EXOC2 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. ab140620 and Anti-GAPDH antibody [6C5] - Loading Control (ab8245) were incubated overnight at 4° at a 1 in 1000 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-EXOC2 antibody [EPR9420] (<a href='/products/primary-antibodies/exoc2-antibody-epr9420-ab140620'>ab140620</a>) at 1/1000 dilution
Lane 1:
Wild-type HeLa cell lysate at 20 µg
Lane 2:
EXOC2 CRISPR/Cas9 edited HeLa cell lysate at 20 µg
Lane 2:
Western blot - Human EXOC2 knockout HeLa cell line (ab265839)
Lane 3:
SH-SY5Y cell lysate at 20 µg
Lane 4:
Daudi cell lysate at 20 µg
Predicted band size: 104 kDa
Observed band size: 104 kDa
false
- Sanger seq
Unknown
Sanger Sequencing - Human EXOC2 knockout HeLa cell line (AB265839)
Allele-1 : 1 bp insertion in exon 3.
- Sanger seq
Unknown
Sanger Sequencing - Human EXOC2 knockout HeLa cell line (AB265839)
Allele-2 : 2 bp insertion in exon 3.
Reactivity data
製品の詳細
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.
製品内容
出荷温度及び保存条件
遺伝子名
遺伝子編集のタイプ
遺伝子編集の方法
ノックアウト検証方法
出荷温度
短期保存温度
長期保存温度
取り扱い方法
初回取り扱いガイドライン
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.
1. Thaw the vial in 37°C water bath for approximately 1-2 minutes.
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.
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.
4. Incubate the culture at 37°C incubator with 5% CO2. Check the culture one day after revival and continue to check until 80% confluent. Media change can be given if needed.
5. Once confluent passage into an appropriate flask at a density of 2x104 cells/cm2. Seeding density is given as a guide only and should be scaled to align with individual lab schedules. Cultures should be monitored daily.
継代培養ガイドライン
- 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.
培養培地
DMEM (High Glucose) + 10% FBS
凍結保存培地
Cell Freezing Medium-DMSO Serum free media, contains 8.7% DMSO in MEM supplemented with methyl cellulose.
補足情報
This supplementary information is collated from multiple sources and compiled automatically.
Biological function summary
EXOC2 acts as a fundamental part of the exocyst complex which comprises eight proteins. This complex orchestrates vesicle targeting and tethering during exocytosis a process essential for maintaining cell surface area and membrane protein composition. Unlike other vesicle trafficking complexes the exocyst shows significant specificity for certain vesicles and localization contributing to the precision of membrane-associated functions.
Pathways
EXOC2 joins key pathways like the insulin signaling pathway and the regulation of actin cytoskeleton. In the insulin signaling pathway EXOC2 integrates with components such as the protein Rab11 to mediate glucose transporter vesicle trafficking impacting glucose uptake. In the regulation of the actin cytoskeleton it interacts with actin-related proteins to influence cell shape and motility.
Quality control
STR analysis
CSF1PO, D13S317, D7S820, D5S818, TH01, D16S539, TPOX
Cell culture
Biosafety level
EU: 2 US: 2
Adherent/suspension
Adherent
Gender
Female
組合せ製品
![Flow Cytometry (Intracellular) - Anti-EXOC2 antibody [EPR9420] (AB140620)](https://content.abcam.com/products/images/exoc2-antibody-epr9420-ab140620--flow-cytometry-intracellular-img37294.jpg)
Abcam 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