Mouse CDKN1B (p27 KIP 1) knockout RAW 264.7 cell line
- Advanced Validation
- What is this?
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- WB
Lab
Western blot - Mouse CDKN1B (p27 KIP 1) knockout RAW 264.7 cell line (AB281619)
False colour image of Western blot : Anti-p27 KIP 1 antibody [EPFHCR16] staining at 1/1000 dilution shown in green; Mouse anti-Alpha Tubulin [DM1A] (ab7291) loading control staining at 1/20000 dilution shown in red. In Western blot ab92741 was shown to bind specifically to p27 KIP 1. A band was observed at 28 kDa in wild-type RAW 264.7 cell lysates with no signal observed at this size in CDKN1B knockout cell line ab281619 (knockout cell lysate ab282970). To generate this image wild-type and CDKN1B knockout RAW 264.7 cell lysates were analysed. First samples were run on an SDS-PAGE gel then transferred onto a nitrocellulose membrane. Membranes were blocked in 3 % milk in TBS-0.1 % Tween® 20 (TBS-T) before incubation with primary antibodies overnight at 4 °. Blots were washed four times in TBS-T incubated with secondary antibodies for 1 h at room temperature washed again four times then imaged. Secondary antibodies used were Goat anti-Rabbit IgG H&L (IRDye® 800CW) preabsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye® 680RD) preabsorbed (ab216776) at 1/20000 dilution.
All lanes:
Western blot - Anti-p27 KIP 1 antibody [EPFHCR16] (<a href='/en-us/products/primary-antibodies/p27-kip-1-antibody-epfhcr16-ab92741'>ab92741</a>) at 1/1000 dilution
Lane 1:
Wild-type RAW 264.7 cell lysate at 20 µg
Lane 2:
CDKN1B knockout RAW 264.7 cell lysate at 20 µg
Lane 2:
Western blot - Mouse CDKN1B (p27 KIP 1) knockout RAW 264.7 cell line (ab281619)
Predicted band size: 22 kDa
Observed band size: 28 kDa
false
- WB
Lab
Western blot - Mouse CDKN1B (p27 KIP 1) knockout RAW 264.7 cell line (AB281619)
False colour image of Western blot : Anti-p27 KIP 1 antibody [Y236] staining at 1/5000 dilution shown in green; Mouse anti-Alpha Tubulin [DM1A] (ab7291) loading control staining at 1/20000 dilution shown in red. In Western blot ab32034 was shown to bind specifically to p27 KIP 1. A band was observed at 28 kDa in wild-type RAW 264.7 cell lysates with no signal observed at this size in CDKN1B knockout cell line ab281619 (knockout cell lysate ab282970). To generate this image wild-type and CDKN1B knockout RAW 264.7 cell lysates were analysed. First samples were run on an SDS-PAGE gel then transferred onto a nitrocellulose membrane. Membranes were blocked in 3 % milk in TBS-0.1 % Tween® 20 (TBS-T) before incubation with primary antibodies overnight at 4 °. Blots were washed four times in TBS-T incubated with secondary antibodies for 1 h at room temperature washed again four times then imaged. Secondary antibodies used were Goat anti-Rabbit IgG H&L (IRDye® 800CW) preabsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye® 680RD) preabsorbed (ab216776) at 1/20000 dilution.
All lanes:
Western blot - Anti-p27 KIP 1 antibody [Y236] (<a href='/en-us/products/primary-antibodies/p27-kip-1-antibody-y236-ab32034'>ab32034</a>) at 1/5000 dilution
Lane 1:
Wild-type RAW 264.7 cell lysate at 20 µg
Lane 2:
CDKN1B knockout RAW 264.7 cell lysate at 20 µg
Lane 2:
Western blot - Mouse CDKN1B (p27 KIP 1) knockout RAW 264.7 cell line (ab281619)
Predicted band size: 22 kDa
Observed band size: 28 kDa
false
- Sanger seq
Supplier Data
Sanger Sequencing - Mouse CDKN1B (p27 KIP 1) knockout RAW 264.7 cell line (AB281619)
52 bp deletion-3 bp insertion in Exon 1
Complementary Products
![Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-p27 KIP 1 antibody [Y236] (AB32034)](https://content.abcam.com/products/images/p27-kip-1-antibody-y236-ab32034--immunohistochemistry-formalin-pfa-fixed-paraffin-embedded-sections-img45173.jpg)
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
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 for bath 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 all remaining cells into an ultra-low attachment 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.
5. Once confluent passage into an appropriate flask at a density of 2x105 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
- Grow in ultra-low attachment flasks.
- Do not wash or use dissociation reagent. Cells are semi-adherent. Tap flask sharply to remove adhered cells.
- All seeding densities should be based on cell counts gained by established methods.
- A guide seeding density of 2x105 cells/mL is recommended.
- Cells should be passaged when they have achieved 80-90% confluence.
Culture medium
DMEM + 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 function of p27 KIP1 involves its role in controlling cell growth and division. It achieves this by becoming a part of larger protein complexes involving CDKs and cyclins. By directly interacting with these complexes p27 KIP1 modulates cell cycle progression therefore acting as a brake on cellular proliferation. The protein is important in maintaining proper cell cycle checkpoints and preventing uncontrolled cell growth which is essential for normal cellular functioning.
Pathways
The involvement of p27 KIP1 centers on cell cycle regulation and signaling pathways such as the PI3K/AKT pathway. Its interaction with CDKs and cyclins situates it within the core mechanisms that determine cell division timing. p27 KIP1 operates alongside other proteins like cyclin D and CDK4/6 fitting into the regulatory intricacies of these pathways. Proper functioning of these pathways ensures cellular homeostasis and prevents the development of oncogenic processes.
Cell culture
Biosafety level
EU: 2 US: 2
Adherent/suspension
Adherent
Gender
Male
Product promise
Please note: All products are 'FOR RESEARCH USE ONLY. NOT FOR USE IN DIAGNOSTIC OR THERAPEUTIC PROCEDURES'.
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