Human IFIH1 knockout Jurkat cell line
- Advanced Validation
- 각 태그의 뜻
Be the first to review this product! 리뷰 제출
|
(0 출판물)
IFIH1 KO cell line available to order. KO validated by Western blot. Free of charge wild type control available. Homozygote, 46 bp deletion. To order both knockout and wild-type control cells: select '2 x 1000000 Cells/vial'. To order only knockout cells: select '1000000 Cells/vial'.
대체 명칭 보기
Murabutide down-regulated protein, Melanoma differentiation-associated protein 5, RH116, RNA helicase-DEAD box protein 116, IFIH1, Interferon-induced with helicase C domain protein 1, Helicard, Interferon-induced helicase C domain-containing protein 1, Clinically amyopathic dermatomyositis autoantigen 140 kDa, CADM-140 autoantigen, MDA5, RLR-2, RIG-I-like receptor 2, MDA-5, Helicase with 2 CARD domains
- WB
Lab
Western blot - Human IFIH1 knockout Jurkat cell line (AB305280)
Western blot : Anti-IFIH1 antibody [EPR6743] (ab126630) 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, ab126630 was shown to bind specifically to IFIH1. A band was observed at 140, 90 kDa in wild-type Jurkat cell lysates with no signal observed at this size in IFIH1 knockout cell line. To generate this image, wild-type and IFIH1 knockout Jurkat 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 800CW and Goat anti-Mouse IgG H&L 680RD at 1/20000 dilution.
All lanes:
Western blot - Anti-MDA5 antibody [EPR6743] (<a href='/ko/products/primary-antibodies/mda5-antibody-epr6743-ab126630'>ab126630</a>) at 1/1000 dilution
Lane 1:
Wild-type Jurkat, LPS (1 ug/mL, 24 h) cell lysate at 20 µg
Lane 2:
Wild-type Jurkat, vehicle control LPS (0 ug/mL, 24 h) cell lysate at 20 µg
Lane 3:
IFIH1 knockout Jurkat, LPS (1 ug/mL, 24 h) cell lysate at 20 µg
Lane 4:
IFIH1 knockout Jurkat, vehicle control LPS (0 ug/mL, 24 h) cell lysate at 20 µg
Predicted band size: 117 kDa,90 kDa
Observed band size: 140 kDa
false
- Sanger seq
Supplier Data
Sanger Sequencing - Human IFIH1 knockout Jurkat cell line (AB305280)
Homozygote, 46 bp deletion
보완 제품
Reactivity 정보
제품 세부 정보
Recommended control: Human wild-type Jurkat cell line (ab275468). Please note a wild-type cell line is not automatically included with a knockout cell line order, if required please add recommended wild-type cell line at no additional cost using the code WILDTYPE-TMTK1.
Cryopreservation cell medium: Cell Freezing Medium-DMSO Serum free media, contains 8.7% DMSO in MEM supplemented with methyl cellulose.
Culture medium: RPMI + 10% FBS
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. Based on cell count, seed cells in an appropriate cell culture flask at a density of 2x105 cells/mL. Seeding density is given as a guide only and should be scaled to align with individual lab schedules.
4. Incubate the culture at 37°C incubator with 5% CO2. 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 2x105 cells/mL is recommended.
• Do not allow the cell density to exceed 3x106 cells/mL
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.
제품 구성
특성 및 보관 정보
유전자명
유전자 편집 종류
유전자 편집 방식
Knockout 검증
배송 시 보관 조건
적절한 단기 보관 기간
적절한 단기 보관 조건
적절한 장기 보관 조건
추가 정보
This supplementary information is collated from multiple sources and compiled automatically.
Biological function summary
In cellular defense mechanisms MDA5 plays an important role in initiating antiviral responses. It operates as part of a signaling complex that includes MAVS (mitochondrial antiviral-signaling protein) leading to the production of type I interferons and pro-inflammatory cytokines. These responses promote an antiviral state in host cells aiding in the containment and clearance of viral infections. MDA5 antibodies can be useful tools for studying these immune processes.
Pathways
MDA5 interacts with antiviral signaling pathways such as the IFN signaling pathway and the RIG-I-like receptor signaling pathway. Upon recognizing viral RNA MDA-5 cooperates with proteins like RIG-I and MAVS to activate downstream signaling cascades that mediate immune responses. This collaboration enables efficient viral recognition and response limiting viral replication and spread.
Cell culture
Biosafety level
EU: 1 US: 1
Adherent/suspension
Suspension
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. <p>1. Thaw the vial in 37°C water for bath approximately 1-2 minutes. <br>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. <br>3. Resuspend the cell pellet in 5 mL pre-warmed culture medium and count using a haemocytometer or alternative cell counting method. Based on cell count, seed cells in an appropriate cell culture flask at a density of 2x10<sup>5</sup> cells/mL. Seeding density is given as a guide only and should be scaled to align with individual lab schedules. <br>4. Incubate the culture at 37°C incubator with 5% CO<sub>2</sub>. Cultures should be monitored daily.</p>
Subculture 가이드라인
- All seeding densities should be based on cell counts gained by established methods.
- A guide seeding density of 2x105 cells/mL is recommended.
- Do not allow cell density to exceed 3x106 cells/mL.
배양 배지
RPMI + 10% FBS
동결보존 배지
Cell Freezing Medium-DMSO Serum free media, contains 8.7% DMSO in MEM supplemented with methyl cellulose.
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