Human HAVCR2 knockout THP-1 cell line
- Advanced Validation
- 각 태그의 뜻
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- WB
Lab
Western blot - Human HAVCR2 knockout THP-1 cell line (AB288696)
Western blot : Anti-TIM 3 antibody [EPR22241] ab241332 staining at 1/1000 dilution, shown in green; Mouse anti GAPDH ab8245 loading control staining at 1/20000 dilution, shown in magenta. A band was observed at 50-60 kDa. To generate this image, samples were run on an SDS-PAGE gel then transferred onto a nitrocellulose membrane. Membranes were blocked in 5% 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 800CW and Goat anti-Mouse 680RD at 1/20,000 dilution.
All lanes:
Western blot - Anti-TIM 3 antibody [EPR22241] (<a href='/ko/products/primary-antibodies/tim-3-antibody-epr22241-ab241332'>ab241332</a>) at 1/1000 dilution
Lane 1:
Wild-type THP-1 vehicle control PMA (0 ng/ml, 48 h) cell lysates at 20 µg
Lane 2:
Western blot - Human HAVCR2 knockout THP-1 cell line (ab288696) at 20 µg
Lane 3:
HAVCR2 knockout THP-1 vehicle control PMA (0 ng/ml, 48 h) cell lysates at 40 µg
Lane 4:
HAVCR2 knockout THP-1 vehicle control PMA (0 ng/ml, 48 h) cell lysates at 20 µg
Lane 5:
HAVCR2 knockout THP-1 treated PMA (100 ng/ml, 48 h) cell lysates at 20 µg
Lane 6:
Daudi cell lysates at 10 µg
Lane 7:
Jurkat cell lysates at 10 µg
Secondary
All lanes:
Goat anti-Rabbit 800CW & Goat anti-Mouse 680RD at 1/20000 dilution
Predicted band size: 33 kDa
Observed band size: 50-60 kDa
false
- Sanger seq
Supplier Data
Sanger Sequencing - Human HAVCR2 knockout THP-1 cell line (AB288696)
Homozygote, 65 bp deletion
보완 제품
Reactivity 정보
제품 세부 정보
Although we aim to provide customers with a homozygous clone, feasibility will be dependent on the biology of the protein. Should only heterozygous edits be achieved, you will be notified of the outcome and be asked to confirm whether the cell line is acceptable. All clones will be accompanied with DNA sequencing data, and the mutation description.
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 조건
적절한 장기 보관 조건
추가 정보
This supplementary information is collated from multiple sources and compiled automatically.
Biological function summary
TIM-3 functions as a checkpoint inhibitor impacting immune cell activity. It is not part of a larger physical complex but it modulates immune responses by interacting with its ligands such as Galectin-9 phosphatidylserine and CEACAM1. TIM-3 involvement in downregulating Th1 cell responses shows its necessary role in maintaining immune homeostasis. The protein also acts in regulating tolerance mechanisms and preventing autoimmunity.
Pathways
TIM-3 participation is seen in the immune checkpoint and T cell exhaustion pathways. TIM-3 signaling results in T cell inhibition affecting the PD-1 pathway as well. It shares a relationship with proteins like LAG-3 and PD-1 which are key to immune inhibitory signaling. These interactions depict TIM-3's role in immune tolerance during chronic infections and malignancies.
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 bath for 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 4x10<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.<br>5. THP-1 cells recover slowly from cryopreservation and therefore may not be ready for subculture for a number of days. Cells should be left as much as possible over this time and only subcultured when the cell density reaches 8x10<sup>5</sup> cells/mL.Small amounts of fresh media can be added until cell number/viability improves.</p>
Subculture 가이드라인
- All seeding densities should be based on cell counts gained by established methods.
- Cells should be seeded at 2x105 - 3x105 cells/mL and subcultured when they have reached 8x105 cells/mL.
- It is not recommended to allow the cell density to exceed 1x106 cells/mL.
배양 배지
RPMI + 10% FBS + 0.05 mM beta-mercaptoethanol
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