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CDKN1B KO cell lysate available now. KO validated by Western blot. Free of charge wild type control included. Knockout achieved by using CRISPR/Cas9,
Homozygous: 52 bp deletion-3 bp insertion in Exon 1.

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Images

Western blot - Mouse CDKN1B (p27 KIP 1) knockout RAW 264.7 cell lysate (AB282970), expandable thumbnail
  • Western blot - Mouse CDKN1B (p27 KIP 1) knockout RAW 264.7 cell lysate (AB282970), expandable thumbnail
  • Sanger Sequencing - Mouse CDKN1B (p27 KIP 1) knockout RAW 264.7 cell lysate (AB282970), expandable thumbnail

Key facts

Cell type

RAW 264.7

Species or organism

Mouse

Tissue

Lymphatic

Knockout validation

Sanger Sequencing, Western blot

Mutation description

Knockout achieved by using CRISPR/Cas9,
Homozygous: 52 bp deletion-3 bp insertion in Exon 1

Alternative names

What's included?

1 Kit
Components
Mouse CDKN1B (p27 KIP 1) knockout RAW 264.7 cell lysate
1 x 100 µg
Mouse wild-type RAW 264.7 cell lysate
1 x 100 µg

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CDKN1B KO cell lysate available now. KO validated by Western blot. Free of charge wild type control included. Knockout achieved by using CRISPR/Cas9,
Homozygous: 52 bp deletion-3 bp insertion in Exon 1.

Alternative names

Key facts

Cell type

RAW 264.7

Mutation description

Knockout achieved by using CRISPR/Cas9,
Homozygous: 52 bp deletion-3 bp insertion in Exon 1

Disease

Carcinoma

Concentration
Loading...

Properties

Gene name

CDKN1B

Gene editing type

Knockout

Gene editing method

CRISPR technology

Knockout validation

Sanger Sequencing, Western blot

Zygosity

Homozygous

Cell culture

Biosafety level

EU: 2 US: 2

Adherent/suspension

Adherent

Gender

Male

Storage

Shipped at conditions

Ambient - Can Ship with Ice

Appropriate long-term storage conditions

-20°C

Notes

Knockout cell lysate achieved by CRISPR/Cas9.

Abcam has not and does not intend to apply for the REACH Authorisation of customers' uses of products that contain European Authorisation list (Annex XIV) substances.
It is the responsibility of our customers to check the necessity of application of REACH Authorisation, and any other relevant authorisations, for their intended uses.

Lysate preparation: Our lysates are made using RIPA buffer to which we add a protease inhibitor cocktail and phosphatase inhibitor cocktail (ratio: 300:100:10). This means that the protein of interest is denatured. If you require a native form of the protein please use the live cell version. Please refer to our lysis protocol for further details on how our lysates are prepared.

User storage instructions: Lyophilizate may be stored at 4°C. After reconstitution, store at -20°C for short-term storage or -80°C for long-term storage.

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.

Supplementary info

Activity summary

The protein known as p27 KIP1 also referred to as p27 CDKN1B or KIP1 is a member of the kinase inhibitory protein family. It plays an important role in cell cycle regulation by inhibiting cyclin-dependent kinases (CDKs). p27 KIP1 weighs approximately 27 kDa and can be found in various tissues where it regulates cell proliferation. The protein functions as a suppressor by binding to cyclin-CDK complexes preventing the transition from G1 phase to S phase in the cell division cycle.

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.

Associated diseases and disorders

Dysfunction of p27 KIP1 has links to cancer and neurodegenerative diseases. Low levels or mutations can lead to uncontrolled cell proliferation contributing to the development and progression of cancers such as breast cancer. p27 KIP1's role in neurodegenerative diseases involves its regulation of neuronal cell cycle re-entry with abnormalities potentially exacerbating conditions like Alzheimer's disease. In these contexts its interaction with proteins such as cyclin E and CDK2 becomes particularly relevant in understanding disease mechanisms.

Product promise

We are dedicated to supporting your work with high quality reagents and we are here for you every step of the way should you need us.

In the unlikely event of one of our products not working as expected, you are covered by our product promise.

Full details and terms and conditions can be found here:
Terms & Conditions.

3 product images

  • Western blot - Mouse CDKN1B (p27 KIP 1) knockout RAW 264.7 cell lysate (ab282970), expandable thumbnail

    Western blot - Mouse CDKN1B (p27 KIP 1) knockout RAW 264.7 cell lysate (ab282970)

    Lane 1: Wild-type RAW 264.7 cell lysate 20 μg
    Lane 2: CDKN1B knockout RAW 264.7 cell lysate 20 μg
    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] (Anti-alpha Tubulin antibody [DM1A] - Loading Control ab7291) loading control staining at 1/20000 dilution, shown in red. In Western blot, Anti-p27 KIP 1 antibody [EPFHCR16] 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 Mouse CDKN1B (p27 KIP 1) knockout RAW 264.7 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 °C. 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 (Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed ab216773) and Goat anti-Mouse IgG H&L (IRDye® 680RD) preabsorbed (Goat anti-Mouse IgG H&L (IRDye® 680RD) preadsorbed ab216776) at 1/20000 dilution.

    All lanes: Western blot - Anti-p27 KIP 1 antibody [EPFHCR16] (Anti-p27 KIP 1 antibody [EPFHCR16] ab92741) 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

    Performed under reducing conditions.

    Predicted band size: 22 kDa

    Observed band size: 28 kDa

  • Western blot - Mouse CDKN1B (p27 KIP 1) knockout RAW 264.7 cell lysate (ab282970), expandable thumbnail

    Western blot - Mouse CDKN1B (p27 KIP 1) knockout RAW 264.7 cell lysate (ab282970)

    Lane 1: Wild-type RAW 264.7 cell lysate 20 μg
    Lane 2: CDKN1B knockout RAW 264.7 cell lysate 20 μg
    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] (Anti-alpha Tubulin antibody [DM1A] - Loading Control ab7291) loading control staining at 1/20000 dilution, shown in red. In Western blot, Anti-p27 KIP 1 antibody [Y236] 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 Mouse CDKN1B (p27 KIP 1) knockout RAW 264.7 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 °C. 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 (Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed ab216773) and Goat anti-Mouse IgG H&L (IRDye® 680RD) preabsorbed (Goat anti-Mouse IgG H&L (IRDye® 680RD) preadsorbed ab216776) at 1/20000 dilution.

    All lanes: Western blot - Anti-p27 KIP 1 antibody [Y236] (Anti-p27 KIP 1 antibody [Y236] ab32034) 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

    Performed under reducing conditions.

    Predicted band size: 22 kDa

    Observed band size: 28 kDa

  • Sanger Sequencing - Mouse CDKN1B (p27 KIP 1) knockout RAW 264.7 cell lysate (ab282970), expandable thumbnail

    Sanger Sequencing - Mouse CDKN1B (p27 KIP 1) knockout RAW 264.7 cell lysate (ab282970)

    52 bp deletion-3 bp insertion in Exon 1

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Product protocols

For this product, it's our understanding that no specific protocols are required. You can:

Please note: All products are 'FOR RESEARCH USE ONLY. NOT FOR USE IN DIAGNOSTIC OR THERAPEUTIC PROCEDURES'.

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