Human ARF1 knockout HeLa cell line
- Advanced Validation
- What is this?
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ARF1 KO cell line available to order. KO validated by Western blot. Free of charge wild type control available. Knockout achieved by using CRISPR/Cas9, 1 bp deletion in exon 2 and 1 bp insertion in exon 2. To order both knockout and wild-type control cells: select '2 x 1000000 Cells/vial'. To order only knockout cells: select '1000000 Cells/vial'.
View Alternative Names
ADP-ribosylation factor 1, ARF1_HUMAN
- WB
Lab
Western blot - Human ARF1 knockout HeLa cell line (AB264939)
Lanes 1 - 4 : Merged signal (red and green). Green - ab183576 observed at 18 kDa. Red - loading control ab7291 (Mouse anti-Alpha Tubulin [DM1A]) observed at 55kDa.
ab183576 was shown to react with ARF1 in wild-type HeLa cells in Western blot with loss of signal observed in ARF1 knockout cell line ab264939 (ARF1 knockout cell lysate ab257353). Wild-type HeLa and ARF1 knockout cell lysates were subjected to SDS-PAGE. Membranes were blocked in 3 % milk in TBS-T (0.1 % Tween®) before incubation with ab183576 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 h at room temperature before imaging.
All lanes:
Western blot - Anti-ARF1 antibody (<a href='/en-us/products/primary-antibodies/arf1-antibody-ab183576'>ab183576</a>) at 1/1000 dilution
Lane 1:
Wild-type HeLa cell lysate at 20 µg
Lane 2:
ARF1 knockout HeLa cell lysate at 20 µg
Lane 2:
Western blot - Human ARF1 knockout HeLa cell line (ab264939)
Lane 3:
MDA-MB-231 cell lysate at 20 µg
Lane 4:
PANC-1 cell lysate at 20 µg
Predicted band size: 21 kDa
Observed band size: 18 kDa
false
- WB
Lab
Western blot - Human ARF1 knockout HeLa cell line (AB264939)
Lanes 1 - 2 : Merged signal (red and green). Green - ab58578 observed at 18 kDa. Red - loading control ab7291 (Mouse anti-Alpha Tubulin [DM1A]) observed at 55 kDa.
ab58578 was shown to react with ARF1 in wild-type HeLa cells in Western blot with loss of signal observed in ARF1 knockout cell line ab264939 (ARF1 knockout cell lysate ab257353). Wild-type HeLa and ARF1 knockout cell lysates were subjected to SDS-PAGE. Membranes were blocked in 3 % milk in TBS-T (0.1 % Tween®) before incubation with ab58578 and ab7291 (Mouse anti-Alpha Tubulin [DM1A]) overnight at 4 °C at 1 μg/ml and a 1 in 20000 dilution respectively. Blots were incubated with Donkey anti-Goat IgG H&L (IRDye® 800CW) preabsorbed (ab216775) and Donkey anti-Mouse 680RD secondary antibodies at 1 in 20000 dilution for 1 h at room temperature before imaging.
All lanes:
Western blot - Anti-ARF1 antibody (<a href='/en-us/products/primary-antibodies/arf1-antibody-ab58578'>ab58578</a>) at 1 µg/mL
Lane 1:
Wild-type HeLa cell lysate at 20 µg
Lane 2:
ARF1 knockout HeLa cell lysate at 20 µg
Lane 2:
Western blot - Human ARF1 knockout HeLa cell line (ab264939)
Predicted band size: 21 kDa
Observed band size: 18 kDa
false
- Sanger seq
Unknown
Sanger Sequencing - Human ARF1 knockout HeLa cell line (AB264939)
Allele-2 : 1 bp insertion in exon 2.
- Sanger seq
Unknown
Sanger Sequencing - Human ARF1 knockout HeLa cell line (AB264939)
Allele-1 : 1 bp deletion in exon 2.
Reactivity data
Product details
We will provide viable cells that proliferate on revival.
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.
What's included?
Properties and storage information
Gene name
Gene editing type
Gene editing method
Knockout validation
Shipped at conditions
Appropriate short-term storage conditions
Appropriate long-term storage conditions
Handling procedures
Initial handling guidelines
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.
Subculture guidelines
- 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.
Culture medium
DMEM (High Glucose) + 10% FBS
Cryopreservation medium
Cell Freezing Medium-DMSO Serum free media, contains 8.7% DMSO in MEM supplemented with methyl cellulose.
Supplementary information
This supplementary information is collated from multiple sources and compiled automatically.
Biological function summary
ARF1 is essential in regulating membrane dynamics and vesicular traffic. It forms part of the COPI and clathrin-coated vesicle complexes where it recruits coat proteins to budding vesicles. This recruitment is fundamental for maintaining Golgi structure and function. Additionally ARF1 plays a role in cytokinesis by interacting with the septin cytoskeleton. The protein also influences actin cytoskeleton remodeling which is pivotal for cell shape changes and motility.
Pathways
ARF1 is central to both the endocytic and secretory pathways. It collaborates with proteins like ARFS and 3F1 in modulating the trafficking of cargo between the endoplasmic reticulum and Golgi. Within the secretory pathway ARF1 interacts with SNARE proteins to facilitate vesicle docking and fusion. Its actions in pathways maintain cellular homeostasis and promote proper cellular response to various stimuli.
Quality control
STR analysis
CSF1PO, D13S317, D7S820, D5S818, TH01, D16S539, TPOX
Cell culture
Biosafety level
EU: 2 US: 2
Adherent/suspension
Adherent
Gender
Female
Complementary Products
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