Human VDR knockout HCT116 cell line
Human VDR knockout HCT116 cell line
- Advanced Validation
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VDR KO cell line available to order. KO validated by Western blot. Free of charge wild type control provided. Knockout.
別名を表示する
1 25 dihydroxyvitamin D3 receptor, 25-dihydroxyvitamin D3 receptor, Member 1, NR1I1, Nuclear receptor subfamily 1 group I member 1, PPP1R163, Protein phosphatase 1, regulatory subunit 163, VDR_HUMAN, Vitamin D (1,25- dihydroxyvitamin D3) receptor, Vitamin D hormone receptor, Vitamin D nuclear receptor variant 1, Vitamin D receptor, Vitamin D3 receptor
- WB
Lab
Western blot - Human VDR knockout HCT116 cell line (AB289181)
Western blot : Anti-VDR antibody [EPR4552] (ab109234) staining at 1/1000 dilution, shown in green; Mouse anti-CANX [CANX/1543] (ab238078) loading control staining at 1/20000 dilution, shown in magenta. In Western blot, ab109234 was shown to bind specifically to VDR. A band was observed at 48 kDa in wild-type HCT 116 cell lysates with no signal observed at this size in VDR knockout cell line. To generate this image, wild-type and VDR knockout HCT 116 cell lysates were analysed. First, 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 IgG H&L 800CW and Goat anti-Mouse IgG H&L 680RD at 1/20000 dilution.
All lanes:
Western blot - Anti-Vitamin D Receptor antibody [EPR4552] - ChIP Grade (<a href='/products/primary-antibodies/vitamin-d-receptor-antibody-epr4552-chip-grade-ab109234'>ab109234</a>) at 1/1000 dilution
Lane 1:
Wild-type HCT 116 cell lysate at 20 µg
Lane 2:
VDR knockout HCT 116 cell lysate at 20 µg
Lane 3:
Wild-type A549 ab288558 cell lysate at 20 µg
Lane 4:
VDR knockout A549 ab288954 cell lysate at 20 µg
Lane 5:
Wild-type HeLa ab255929 cell lysate at 20 µg
Lane 6:
VDR knockout HeLa <a href='/products/cell-lines/human-vdr-vitamin-d-receptor-knockout-hela-cell-line-ab265430'>ab265430</a> cell lysate at 20 µg
Secondary
All lanes:
Goat anti-Rabbit IgG H&L 800CW and Goat anti-Mouse IgG H&L 680RD at 1/20000 dilution
Observed band size: 48 kDa
false
Reactivity data
製品の詳細
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.
Recommended control: Human wild-type HCT116 cell line (ab288559). 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.
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.
出荷温度及び保存条件
遺伝子名
遺伝子編集のタイプ
遺伝子編集の方法
ノックアウト検証方法
出荷温度
短期保存温度
長期保存温度
取り扱い方法
初回取り扱いガイドライン
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 bath for 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 seed all remaining cells into a 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. Media change can be given if needed.
5. Once confluent passage into an appropriate flask at a density of 2x104 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.
継代培養ガイドライン
- All seeding densities should be based on cell counts gained by established methods.
- A guide seeding density of 2x104 cells/cm2 is recommended.
- Cells should be passaged when they have achieved 80-90% confluence.
培養培地
McCoY5a + 10% FBS
凍結保存培地
Cell Freezing Medium-DMSO Serum free media, contains 8.7% DMSO in MEM supplemented with methyl cellulose.
Quality control
STR analysis
D5S818, TH01, D16S539, TPOX, CSF1PO, D13S317, D7S820
Cell culture
Biosafety level
EU: 1 US: 1
Adherent/suspension
Adherent
Gender
Male
ターゲットの情報
Complementary Products
Primary Antibodies
AB109234
Anti-Vitamin D Receptor antibody [EPR4552] - ChIP Grade
RabMAb|Recombinant|KO Validated
![Western blot - Anti-Vitamin D Receptor antibody [EPR4552] - ChIP Grade (AB109234)](https://content.abcam.com/products/images/vitamin-d-receptor-antibody-epr4552-chip-grade-ab109234--western-blot-img436354.jpg)
- 0
- 0
![Western blot - Anti-CYP27B1 antibody [EPR20271] (AB206655)](https://content.abcam.com/products/images/cyp27b1-antibody-epr20271-ab206655--western-blot-img17509.jpg)
Cell Lines & Lysates
AB265557
Human POLM (DNA polymerase mu) knockout HeLa cell line
Advanced Validation
- 0
- 0
Abcam product promise
Please note: All products are 'FOR RESEARCH USE ONLY. NOT FOR USE IN DIAGNOSTIC OR THERAPEUTIC PROCEDURES'.
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