Human GRN (Granulin) knockout HEK-293T cell line
- Advanced Validation
- What is this?
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GRN KO cell line available to order. KO validated by Western blot. Free of charge wild type control available. Knockout achieved by using CRISPR/Cas9, 14 bp deletion in exon 2 and 2 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
Acrogranin, CLN11, GEP, GP88, GRN_HUMAN, Granulin A, Granulin B, Granulin C, Granulin D, Granulin E, Granulin F, Granulin G, Granulin epithelin, Granulin-7, Granulins, PC cell derived growth factor, PCDGF, PEPI, PGRN, Proepithelin, Progranulin
- WB
Lab
Western blot - Human GRN (Granulin) knockout HEK-293T cell line (AB266738)
Lanes 1-4 : Merged signal (red and green). Green - ab208777 observed at 74 kDa. Red - loading control ab8245 observed at 36 kDa.
ab208777 Anti-Granulin antibody [EPR15864] was shown to specifically react with Granulin in wild-type HEK293T cells. Loss of signal was observed when knockout cell line ab266738 (knockout cell lysate ab257235) was used. Wild-type and Granulin knockout samples were subjected to SDS-PAGE. ab208777 and Anti-GAPDH antibody [6C5] - Loading Control (ab8245) were incubated at room temperature for 2.5 hours 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-Granulin antibody [EPR15864] (<a href='/en-us/products/primary-antibodies/granulin-antibody-epr15864-ab208777'>ab208777</a>) at 1/1000 dilution
Lane 1:
Wild-type HEK293T cell lysate at 20 µg
Lane 2:
GRN knockout HEK293T cell lysate at 20 µg
Lane 2:
Western blot - Human GRN (Granulin) knockout HEK-293T cell line (ab266738)
Lane 3:
HepG2 cell lysate at 20 µg
Lane 4:
HAP1 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: 64 kDa
Observed band size: 74 kDa
false
- Sanger seq
Unknown
Sanger Sequencing - Human GRN (Granulin) knockout HEK-293T cell line (AB266738)
Allele-1 : 14 bp deletion in exon2
- Cell Culture
Unknown
Cell Culture - Human GRN (Granulin) knockout HEK-293T cell line (AB266738)
Representative images of GRN 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 an EVOS M5000 microscope.
- Sanger seq
Unknown
Sanger Sequencing - Human GRN (Granulin) knockout HEK-293T cell line (AB266738)
Allele-2 : 2 bp deletion in exon 2.
- WB
Lab
Western blot - Human GRN (Granulin) knockout HEK-293T cell line (AB266738)
All lanes:
Western blot - Anti-Granulin antibody [EPR15864] (<a href='/en-us/products/primary-antibodies/granulin-antibody-epr15864-ab208777'>ab208777</a>) at 1/1000 dilution
Lane 1:
Wild-type HEK293T lysate at 20 µg
Lane 2:
Western blot - Human GRN (Granulin) knockout HEK-293T cell line (ab266738)
Lane 2:
Western blot - Human GRN (Granulin) knockout HEK-293T cell lysate (<a href='/en-us/products/cell-lysates/human-grn-granulin-knockout-hek-293t-cell-lysate-ab257235'>ab257235</a>)
Observed band size: 64 kDa
false
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
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
Granulin functions as a growth factor involved in tissue repair cell proliferation and inflammation. It does not function as part of a larger protein complex but acts individually in exerting its effects on cells. Granulin influences various aspects of cellular architecture and signaling pathways making it important for maintaining cellular homeostasis. Additionally the granulin protein participates in regulating networks that involve cytokines and growth factors impacting both the innate and adaptive immune responses.
Pathways
Granulin plays significant roles in the MAPK and NF-kB signaling pathways. These pathways critically manage cellular reactions to stress inflammation and other external stimuli. Granulin modulates these processes often interacting with proteins like TNF receptors and interleukins to affect the downstream signaling events. Interactions between granulin and other signaling molecules allow it to integrate into broader networks that oversee various biological responses.
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
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Advanced Validation
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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