Human A-Raf knockout HEK-293T cell line
- Advanced Validation
- What is this?
Be the first to review this product! Submit a review
|
(0 Publication)
ARAF KO cell line available to order. KO validated by Western blot. Free of charge wild type control available. Knockout achieved by using CRISPR/Cas9, Homozygous: 1 bp deletion in exon 2. To order both knockout and wild-type control cells: select 2 x 1000000Cells/vial. To order only knockout cells: select 1000000Cells/vial.
View Alternative Names
A Raf proto oncogene serine/threonine protein kinase, A raf 1, ARAF_HUMAN, Oncogene Araf1, Oncogene PKS2, PKS, PKS 2, Proto-oncogene A-Raf, Proto-oncogene A-Raf-1, Proto-oncogene Pks, RAFA 1, Ras binding protein DA Raf, Serine/threonine-protein kinase A-Raf, v raf murine sarcoma 3611 viral oncogene homolog, v raf murine sarcoma 3611 viral oncogene homolog 1, v raf oncogene homolog 1 (murine sarcoma 3611 virus)
- WB
Lab
Western blot - Human A-Raf knockout HEK-293T cell line (AB266351)
Lanes 1-4 : Merged signal (red and green). Green - ab200653 observed at 68 kDa. Red - loading control ab8245 observed at 36 kDa.
ab200653 Anti-A-Raf antibody [EPR16208] was shown to specifically react with A-Raf in wild-type HEK-293T cells. Loss of signal was observed when knockout cell line ab266351 (knockout cell lysate ab257838) was used. Wild-type and A-Raf knockout samples were subjected to SDS-PAGE. ab200653 and Anti-GAPDH antibody [6C5] - Loading Control (ab8245) were incubated overnight at 4° at 1 in 1000 dilution and 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-A-Raf antibody [EPR16208] (<a href='/en-us/products/primary-antibodies/a-raf-antibody-epr16208-ab200653'>ab200653</a>) at 1/1000 dilution
Lane 1:
Wild-type HEK-293T (Human epithelial cell line from embryonic kidney transformed with large T antigen) whole cell lysate at 20 µg
Lane 2:
A-Raf knockout HEK-293T (Human epithelial cell line from embryonic kidney transformed with large T antigen) whole cell lysate at 20 µg
Lane 2:
Western blot - Human A-Raf knockout HEK-293T cell line (ab266351)
Lane 3:
HeLa (Human epithelial cell line from cervix adenocarcinoma) whole cell lysate at 20 µg
Lane 4:
Raji (Human Burkitts lymphoma cell line) whole cell lysate at 20 µg
Secondary
All lanes:
Western blot - Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed (<a href='/en-us/products/secondary-antibodies/goat-rabbit-igg-h-l-irdye-800cw-preadsorbed-ab216773'>ab216773</a>) at 1/10000 dilution
Predicted band size: 68 kDa
Observed band size: 68 kDa
false
- Sanger seq
Unknown
Sanger Sequencing - Human A-Raf knockout HEK-293T cell line (AB266351)
Homozygous : 1 bp deletion in exon 2
- Cell Culture
Lab
Cell Culture - Human A-Raf knockout HEK-293T cell line (AB266351)
Representative images A-Raf knockout HEK293T cells, low and high confluency examples (top left and right respectively) and wild-type HEK293T cells, low and high confluency (bottom left and right respectively) showing typical adherent, epithelial-like morphology. Images were captured at 10X magnification using a EVOS M5000 microscope.
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 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
The A-Raf protein participates in the regulation of cell growth and survival. It forms part of the MAPK/ERK signaling cascade which is essential for transmitting mitogenic signals from the cell surface to the nucleus. A-Raf acts as a modulator within this complex impacting the overall signaling output. The protein is intimately involved in the regulation of the developmental processes influencing cell fate and function.
Pathways
The A-Raf protein is central to the MAPK/ERK pathway a critical signaling pathway involved in the regulation of cellular responses to growth signals. This pathway includes key components like the proteins B-Raf and C-Raf with A-Raf playing specific roles in the modulation and fine-tuning of this signal transmission. It helps mediate signals that control cell division differentiation and secretion ensuring appropriate cellular responses to external 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
![Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-A-Raf antibody [EPR16208] (AB200653)](https://content.abcam.com/products/images/a-raf-antibody-epr16208-ab200653--immunohistochemistry-formalin-pfa-fixed-paraffin-embedded-sections-img107369.jpg)
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