Human RAB27A knockout A549 cell line
- Advanced Validation
- What is this?
Be the first to review this product! Submit a review
|
(0 Publication)
RAB27A KO cell line available to order. KO validated by Western blot. Free of charge wild type control provided. Knockout achieved by using CRISPR/Cas9, 16 bp deletion in exon 5 and 2 bp deletion in exon 5 and 5 bp deletion in exon 5 and 7 bp insertion in exon 5.
View Alternative Names
GS2, GTP-binding protein Ram, HsT18676, MGC117246, Mutant Ras related protein Rab-27A, RAB-27A, RAB27A member RAS oncogene family, RAM, RB27A_HUMAN, Rab-27, Ras-related protein Rab-27A
- WB
Lab
Western blot - Human RAB27A knockout A549 cell line (AB266921)
Lanes 1 - 2 : Merged signal (red and green). Green - anti-RAB27A antibody observed at 140 kDa. Red - loading control ab8245 (Mouse anti-GAPDH antibody [6C5]) observed at 37kDa.
anti-RAB27A was shown to react with RAB27A in wild-type A549 cells in Western blot with loss of signal observed in RAB27A knockout cell line ab266921 (RAB27A knockout cell lysate ab258618). Wild-type A549 and RAB27A knockout cell lysates were subjected to SDS-PAGE. Membranes were blocked in fluorescent western blot (TBS-based) blocking solution before incubation with anti-RAB27A antibody 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.
All lanes:
anti-RAB27A antibody at 1/1000 dilution
Lanes 1 - 2:
RAB27A knockout A549 cell lysate at 40 µg
Lane 2:
Western blot - Human RAB27A knockout A549 cell line (ab266921)
false
- Sanger seq
Unknown
Sanger Sequencing - Human RAB27A knockout A549 cell line (AB266921)
Allele-1 : 16 bp deletion in exon5
- Sanger seq
Unknown
Sanger Sequencing - Human RAB27A knockout A549 cell line (AB266921)
Allele-3 : 2 bp deletion in exon 5.
- Sanger seq
Unknown
Sanger Sequencing - Human RAB27A knockout A549 cell line (AB266921)
Allele-2 : 5 bp deletion in exon 5.
Reactivity data
Product details
Recommended control: Human wild-type A549 cell line (ab255450). 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, 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.
- Do not allow the cell density to exceed 7x104 cells/cm2.
Culture medium
F-12K + 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
This protein regulates the exocytosis processes in certain secretory cells. RAB27A partners with effectors through its GDP-bound and GTP-bound states forming protein complexes to mediate the movement and anchoring of secretory granules. Melanocytes depend on RAB27A to control the distribution of melanosomes which are organelles containing pigment melanin. In immune cells RAB27A regulates the secretion of lytic granules significantly impacting the immune response.
Pathways
This protein is involved in the melanogenesis pathway and the lytic granule exocytosis pathway. In the melanogenesis pathway RAB27A works closely with proteins such as Melanophilin and Myosin Va to control melanosome transport. In the exocytosis pathway in immune cells RAB27A interacts with proteins like UNC13D which are vital for the release of cytotoxic molecules. By controlling these pathways RAB27A influences pigment formation in skin and hair as well as immune defense mechanisms.
Quality control
STR analysis
CSF1PO, D13S317, D7S820, D5S818, TH01, D16S539, TPOX
Cell culture
Biosafety level
EU: 1 US: 1
Adherent/suspension
Adherent
Gender
Male
Target data
Complementary Products
Primary Antibodies
AB313575
Anti-RAB27A + RAB27B antibody [EPR27120-68]
BOND RX™ Validated|20ul selling size|RabMAb|Recombinant
![Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-RAB27A + RAB27B antibody [EPR27120-68] (AB313575)](https://content.abcam.com/products/images/rab27a-rab27b-antibody-epr27120-68-ab313575--immunohistochemistry-formalin-pfa-fixed-paraffin-embedded-sections-img418683.jpg)
- 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