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AB261743

Human FANCD2 knockout HeLa cell line

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FANCD2 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: 4 bp deletion in exon 13. 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
Western blot - Human FANCD2 knockout HeLa cell line (AB261743)
  • WB

Lab

Western blot - Human FANCD2 knockout HeLa cell line (AB261743)

Lanes 1- 2 : Merged signal (red and green). Green - ab178705 observed at 166 kDa. Red - Anti-alpha Tubulin antibody [DM1A] - Loading Control (ab7291) observed at 50 kDa.

ab178705 was shown to react with FANCD2 in wild-type HeLa cells in western blot. Loss of signal was observed when knockout cell line ab261743 (knockout cell lysate ab257173) was used. Wild-type HeLa and FANCD2 knockout HeLa cell lysates were subjected to SDS-PAGE. Membrane was blocked for 1 hour at room temperature in 0.1% TBST with 3% non-fat dried milk. ab178705 and Anti-alpha Tubulin antibody [DM1A] - Loading Control (ab7291) overnight at 4°C at a 1 in 1000 dilution and a 1 in 20000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye®800CW) preadsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye®680RD) preadsorbed (ab216776) secondary antibodies at 1 in 20000 dilution for 1 hour at room temperature before imaging.

All lanes:

Western blot - Anti-FANCD2 antibody [EPR2279(3)] (<a href='/ko/products/primary-antibodies/fancd2-antibody-epr22793-ab178705'>ab178705</a>) at 1/1000 dilution

Lane 1:

Wild-type HeLa cell lysate at 20 µg

Lane 2:

FANCD2 knockout HeLa cell lysate at 20 µg

Lane 2:

Western blot - Human FANCD2 knockout HeLa cell line (ab261743)

Predicted band size: 166 kDa

Observed band size: 166 kDa

false

Western blot - Human FANCD2 knockout HeLa cell line (AB261743)
  • WB

Lab

Western blot - Human FANCD2 knockout HeLa cell line (AB261743)

Lanes 1- 2 : Merged signal (red and green). Green - ab108928 observed at 166 kDa. Red - Anti-alpha Tubulin antibody [DM1A] - Loading Control (ab7291) observed at 50 kDa.

ab108928 was shown to react with FANCD2 in wild-type HeLa cells in western blot. Loss of signal was observed when knockout cell line ab261743 (knockout cell lysate ab257173) was used. Wild-type HeLa and FANCD2 knockout HeLa cell lysates were subjected to SDS-PAGE. Membrane was blocked for 1 hour at room temperature in 0.1% TBST with 3% non-fat dried milk. ab108928 and Anti-alpha Tubulin antibody [DM1A] - Loading Control (ab7291) overnight at 4°C at a 1 in 1000 dilution and a 1 in 20000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye®800CW) preadsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye®680RD) preadsorbed (ab216776) secondary antibodies at 1 in 20000 dilution for 1 hour at room temperature before imaging.

All lanes:

Western blot - Anti-FANCD2 antibody [EPR2302] (<a href='/ko/products/primary-antibodies/fancd2-antibody-epr2302-ab108928'>ab108928</a>) at 1/1000 dilution

Lane 1:

Wild-type HeLa cell lysate at 20 µg

Lane 2:

FANCD2 knockout HeLa cell lysate at 20 µg

Lane 2:

Western blot - Human FANCD2 knockout HeLa cell line (ab261743)

Predicted band size: 166 kDa

Observed band size: 166 kDa

false

Sanger Sequencing - Human FANCD2 knockout HeLa cell line (AB261743)
  • Sanger seq

Unknown

Sanger Sequencing - Human FANCD2 knockout HeLa cell line (AB261743)

Homozygous : 4 bp deletion in exon 13.

주요 정보

세포 유형

HeLa

Species or organism

Human

조직

Cervix

제형

Liquid

form

Knockout 검증

Sanger Sequencing,Western blot

Mutation 정보

Knockout achieved by using CRISPR/Cas9, Homozygous: 4 bp deletion in exon 13

Antibiotic resistance

Puromycin 1µg/mL

질병

Adenocarcinoma

Reactivity 정보

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

제품 세부 정보

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

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특성 및 보관 정보

유전자명
FANCD2
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

추가 정보

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

FANCD2 also known as Fanconi anemia group D2 functions mechanically in DNA repair processes. This protein plays a role in the cellular response to DNA damage specifically interstrand crosslinks. FANCD2 has a molecular weight of approximately 164 kDa. Expression of FANCD2 occurs broadly in various tissues but reaches higher levels in cells that undergo rapid division such as hematopoietic cells.
Biological function summary

FANCD2 collaborates with multiple proteins as part of the Fanconi anemia (FA) complex to ensure genomic stability. The FA complex operates by facilitating the repair of DNA interstrand crosslinks that impede replication. FANCD2 becomes activated through monoubiquitination which serves as a signal for recruitment to DNA damage sites. It essentially functions as a coordinator that brings together important components for repair.

Pathways

FANCD2 integrates into the Fanconi anemia pathway and the homologous recombination repair pathway. These pathways are critical for maintaining DNA integrity and preventing chromosomal instability. Within these pathways interactions with proteins such as BRCA1 and BRCA2 reinforce the repair processes. FANCD2's connection to these proteins exemplifies its role in complex mechanisms that preserve genomic fidelity.

FANCD2 links significantly to Fanconi anemia a genetic disorder that causes bone marrow failure and increased cancer susceptibility. Damage or malfunction of FANCD2 can disrupt DNA repair leading to cellular dysfunction seen in this condition. Additionally there is a connection to breast cancer whereby FANCD2 interacts with BRCA2 indicating a shared pathway involved in tumor suppressor functions. These associations underline the importance of FANCD2 in disease pathology and its potential as a therapeutic target.

품질 관리

STR analysis

CSF1PO, D13S317, D7S820, D5S818, TH01, D16S539, TPOX

Cell culture

Biosafety level

EU: 2 US: 2

Adherent/suspension

Adherent

Gender

Female

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 seed all remaining cells into a T25.<br>4. Incubate the culture at 37°C incubator with 5% CO<sub>2</sub>. Check the culture one day after revival and continue to check until 80% confluent. Media change can be given if needed.<br>5. Once confluent passage into an appropriate flask at a density of 2x10<sup>4</sup> cells/cm<sup>2</sup>. Seeding density is given as a guide only and should be scaled to align with individual lab schedules. 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 2x104 cells/cm2 is recommended.
  • Cells should be passaged when they have achieved 80-90% confluence.
배양 배지

DMEM (High Glucose) + 10% FBS

동결보존 배지

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

제품 프로토콜

Product promise

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