AB281619

Mouse CDKN1B (p27 KIP 1) knockout RAW 264.7 cell line

Advanced Validation
각 태그의 뜻

Be the first to review this product! 리뷰 제출

(0 출판물)

CDKN1B 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: 52 bp deletion-3 bp insertion in Exon 1. To order both knockout and wild-type control cells: select '2 x 1000000 Cells/vial'. To order only knockout cells: select '1000000 Cells/vial'.

3 이미지

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='/ko/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

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='/ko/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 Sequencing - Mouse CDKN1B (p27 KIP 1) knockout RAW 264.7 cell line (AB281619)

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

주요 정보

세포 유형

RAW 264.7

Species or organism

Mouse

조직

Lymphatic

제형

Liquid

form

Knockout 검증

Sanger Sequencing,Western blot

Mutation 정보

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

질병

Carcinoma

style

left-column

style

left-column

Reactivity 정보

{ "title": "Reactivity Data", "filters": { "stats": ["", "Reactivity", "Dilution Info", "Notes"] }, "values": { "WB": { "reactivity":"TESTED_AND_REACTS", "dilution-info":"", "notes":"" } } }

style

left-column

제품 세부 정보

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.

style

left-column

제품 구성

{ "values": { "2x1000000Cellsvial": { "sellingSize": "2 x 1000000 Cells/vial", "publicAssetCode":"ab281619-2x1000000Cells_vial", "assetComponentDetails": [ { "size":"1 x 1000000 Cells/vial", "name":"ab275474 Mouse wild-type RAW 264.7 cell line", "number":"AB281619-CMP02" }, { "size":"1 x 1000000 Cells/vial", "name":"ab281619 Mouse CDKN1B (p27 KIP 1) knockout RAW 264.7 cell line", "number":"AB281619-CMP01" } ] }, "1000000Cellsvial": { "sellingSize": "1000000 Cells/vial", "publicAssetCode":"ab281619-1000000Cells_vial", "assetComponentDetails": [ { "size":"1 x 1000000 Cells/vial", "name":"ab281619 Mouse CDKN1B (p27 KIP 1) knockout RAW 264.7 cell line", "number":"AB281619-CMP01", "productcode":"" } ] } } }

style

left-column

특성 및 보관 정보

유전자명

CDKN1B

유전자 편집 종류

Knockout

유전자 편집 방식

CRISPR technology

Knockout 검증

Sanger Sequencing, Western blot

Zygosity

Homozygous

배송 시 보관 조건

Dry Ice

적절한 단기 보관 조건

-196°C

적절한 장기 보관 조건

-196°C

style

left-column

추가 정보

This supplementary information is collated from multiple sources and compiled automatically.

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.

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.

style

left-column

style

left-column

Cell culture

Biosafety level

EU: 2 US: 2

Adherent/suspension

Adherent

Gender

Male

초기 취급 지침

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 가이드라인

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.

배양 배지

DMEM + 10% FBS

동결보존 배지

Cell Freezing Medium-DMSO Serum free media, contains 8.7% DMSO in MEM supplemented with methyl cellulose.

style

left-column

제품 프로토콜

Visit the General protocols
Visit the Troubleshooting

style

left-column

style

left-column

style

right-column

Product promise

저희는 고품질 시약으로 고객님의 연구를 서포트하고자 최선을 다하고 있으며, 모든 단계에서 함께 하겠습니다. 만약 제품이 기대한 성능을 보이지 않을 경우에도 Product Promise로 보호받으실 수 있습니다.

자세한 내용은 이용 약관을 확인해 주세요.

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

Mouse CDKN1B (p27 KIP 1) knockout RAW 264.7 cell line

Western blot - Mouse CDKN1B (p27 KIP 1) knockout RAW 264.7 cell line (AB281619)

Western blot - Mouse CDKN1B (p27 KIP 1) knockout RAW 264.7 cell line (AB281619)

Sanger Sequencing - Mouse CDKN1B (p27 KIP 1) knockout RAW 264.7 cell line (AB281619)

주요 정보

세포 유형

Species or organism

조직

제형

Knockout 검증

Mutation 정보

질병

보완 제품

Primary Antibodies

Primary Antibodies

Primary Antibodies

Primary Antibodies

Reactivity 정보

제품 세부 정보

제품 구성

특성 및 보관 정보

유전자명

유전자 편집 종류

유전자 편집 방식

Knockout 검증

Zygosity

배송 시 보관 조건

적절한 단기 보관 조건

적절한 장기 보관 조건

추가 정보

Biological function summary

Pathways

Cell culture

Biosafety level

Adherent/suspension

Gender

초기 취급 지침

Subculture 가이드라인

배양 배지

동결보존 배지

제품 프로토콜

Product promise