Human RAB7A (RAB7) knockout HeLa cell line
- Advanced Validation
- What is this?
Be the first to review this product! Submit a review
|
(1 Publication)
RAB7A KO cell line available to order. KO validated by Western blot. Free of charge wild type control provided. Knockout achieved by using CRISPR/Cas9, Homozygous: 25 bp deletion in exon 2.
View Alternative Names
CMT2B, PRO2706, PSN, RAB7, member RAS oncogene family, RAB7A, member RAS oncogene family, RAB7A_HUMAN, Ras associated protein RAB7, Ras related protein Rab7, Ras-related protein Rab-7a
- WB
Lab
Western blot - Human RAB7A (RAB7) knockout HeLa cell line (AB255423)
Lanes 1 - 2 : Merged signal (red and green). Green - ab126712 observed at 23 kDa. Red - loading control ab7291 (Mouse anti-Alpha Tubulin [DM1A]) observed at 55 kDa.
ab126712 was shown to react with Rab7 in wild-type HeLa cells in Western blot with loss of signal observed in Rab7A knockout cell line ab255423 (Rab7A knockout cell lysate ab263831). Wild-type HeLa and Rab7A knockout cell lysates were subjected to SDS-PAGE. Membranes were blocked in 3 % milk in TBS-T (0.1 % Tween®) before incubation with ab126712 and ab7291 (Mouse anti-Alpha Tubulin [DM1A]) overnight at 4 ° 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-RAB7 antibody [EPR7588(B)] - Late Endosome Marker (<a href='/en-us/products/primary-antibodies/rab7-antibody-epr7588b-late-endosome-marker-ab126712'>ab126712</a>) at 1/1000 dilution
Lane 1:
Wild-type HeLa cell lysate at 20 µg
Lane 2:
RAB7A knockout HeLa cell lysate at 20 µg
Lane 2:
Western blot - Human RAB7A (RAB7) knockout HeLa cell line (ab255423)
Observed band size: 23 kDa
false
- WB
Lab
Western blot - Human RAB7A (RAB7) knockout HeLa cell line (AB255423)
Lanes 1 - 2 : Merged signal (red and green). Green - ab137029 observed at 23 kDa. Red - loading control ab7291 (Mouse anti-Alpha Tubulin [DM1A]) observed at 55 kDa.
ab137029 was shown to react with Rab7 in wild-type HeLa cells in Western blot with loss of signal observed in Rab7A knockout cell line ab255423 (Rab7A knockout cell lysate ab263831). Wild-type HeLa and Rab7A knockout cell lysates were subjected to SDS-PAGE. Membranes were blocked in 3 % milk in TBS-T (0.1 % Tween®) before incubation with ab137029 and ab7291 (Mouse anti-Alpha Tubulin [DM1A]) overnight at 4 ° 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-RAB7 antibody [EPR7589] - Late Endosome Marker (<a href='/en-us/products/primary-antibodies/rab7-antibody-epr7589-late-endosome-marker-ab137029'>ab137029</a>) at 1/1000 dilution
Lane 1:
Wild-type HeLa cell lysate at 20 µg
Lane 2:
RAB7A knockout HeLa cell lysate at 20 µg
Lane 2:
Western blot - Human RAB7A (RAB7) knockout HeLa cell line (ab255423)
Observed band size: 23 kDa
false
- Sanger seq
Unknown
Sanger Sequencing - Human RAB7A (RAB7) knockout HeLa cell line (AB255423)
Homozygous : 25 bp deletion in exon 2.
Reactivity data
Product details
Recommended control: Human wild-type HeLa cell line (ab255928). Please note a wild-type cell line is not automatically included with a knockout cell line order, if required please add recommended wild-type cell line at no additional cost using the code WILDTYPE-TMTK1.
We will provide viable cells that proliferate on revival.
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.
What's included?
Properties and storage information
Gene name
Gene editing type
Gene editing method
Knockout validation
Zygosity
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
RAB7 affects various processes including autophagy cell signaling and pathogen infection response. It often functions as part of the endosomal machinery and forms complexes with other proteins like RAB3GAP1 and RILP. These complexes help mediate vesicle transport and membrane trafficking processes which are essential for the maintenance of cellular homeostasis. RAB7 is significant in ensuring proper lysosomal positioning and function which is critical for cellular metabolism and degradation.
Pathways
RAB7 integrates into the endocytic and autophagic pathways by facilitating the transport between endosomes and lysosomes. It connects with the PI3K/AKT pathway impacting cell proliferation and survival. RAB7 also interacts with the retromer complex influencing the retrieval of receptors from endosomes. Proteins like FYCO1 and Prostaglandin E2 synthase can also cooperate with RAB7 in these pathways further establishing its role in cellular dynamics and signaling.
Quality control
STR analysis
CSF1PO, D13S317, D7S820, D5S818, TH01, D16S539, TPOX
Cell culture
Biosafety level
EU: 2 US: 2
Adherent/suspension
Adherent
Gender
Female
Target data
Publications (1)
Recent publications for all applications. Explore the full list and refine your search
Stem cells international 2021:9992381 PubMed34367295
2021
Applications
Unspecified application
Species
Unspecified reactive species
Complementary Products
Cell Lines & Lysates
AB255371
Human CAV1 (Caveolin-1) knockout HeLa cell line
Advanced Validation
- 0
- 0
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