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AB265946

Human NDUFB9 knockout HeLa cell line

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NDUFB9 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: 1 bp insertion in exon 2. 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 NDUFB9 knockout HeLa cell line (AB265946)
  • WB

Lab

Western blot - Human NDUFB9 knockout HeLa cell line (AB265946)

Lanes 1-3 : Merged signal (red and green). Green - ab200198 observed at 22 kDa. Red - loading control ab8245 observed at 36 kDa.

ab200198 Anti-NDUFB9 antibody [EPR15955-78] was shown to specifically react with NDUFB9 in wild-type HeLa cells. Loss of signal was observed when knockout cell line ab265946 (knockout cell lysate ab258065) was used. Wild-type and NDUFB9 knockout samples were subjected to SDS-PAGE. ab200198 and Anti-GAPDH antibody [6C5] - Loading Control (ab8245) were incubated overnight at 4° at 1 in 1000 dilution and 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-NDUFB9 antibody [EPR15955-78] (<a href='/ko/products/primary-antibodies/ndufb9-antibody-epr15955-78-ab200198'>ab200198</a>) at 1/1000 dilution

Lane 1:

Wild-type HeLa cell lysate at 20 µg

Lane 2:

NDUFB9 knockout HeLa cell lysate at 20 µg

Lane 2:

Western blot - Human NDUFB9 knockout HeLa cell line (ab265946)

Lane 3:

Jurkat cell lysate at 20 µg

Secondary

All lanes:

Western blot - Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed (<a href='/ko/products/secondary-antibodies/goat-rabbit-igg-h-l-irdye-800cw-preadsorbed-ab216773'>ab216773</a>) at 1/10000 dilution

Predicted band size: 21 kDa

Observed band size: 22 kDa

false

Western blot - Human NDUFB9 knockout HeLa cell line (AB265946)
  • WB

Unknown

Western blot - Human NDUFB9 knockout HeLa cell line (AB265946)

Lanes 1-3 : Merged signal (red and green). Green - ab188581 observed at 22 kDa. Red - loading control ab8245 observed at 36 kDa.

ab188581 Anti-NDUFB9 antibody [EPR15955] was shown to specifically react with NDUFB9 in wild-type HeLa cells. Loss of signal was observed when knockout cell line ab265946 (knockout cell lysate ab258065) was used. Wild-type and NDUFB9 knockout samples were subjected to SDS-PAGE. ab188581 and Anti-GAPDH antibody [6C5] - Loading Control (ab8245) were incubated overnight at 4° at 1 in 1000 dilution and 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-NDUFB9 antibody [EPR15955] (<a href='/ko/products/primary-antibodies/ndufb9-antibody-epr15955-ab188581'>ab188581</a>) at 1/1000 dilution

Lane 1:

Wild-type HeLa cell lysate at 20 µg

Lane 2:

NDUFB9 knockout HeLa cell lysate at 20 µg

Lane 2:

Western blot - Human NDUFB9 knockout HeLa cell line (ab265946)

Lane 3:

Jurkat cell lysate at 20 µg

Secondary

All lanes:

Western blot - Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed (<a href='/ko/products/secondary-antibodies/goat-rabbit-igg-h-l-irdye-800cw-preadsorbed-ab216773'>ab216773</a>) at 1/10000 dilution

Predicted band size: 21 kDa

Observed band size: 22 kDa

false

Sanger Sequencing - Human NDUFB9 knockout HeLa cell line (AB265946)
  • Sanger seq

Unknown

Sanger Sequencing - Human NDUFB9 knockout HeLa cell line (AB265946)

Homozygous : 1 bp insertion in exon 2.

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: 1 bp insertion in exon 2

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, 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.

제품 구성

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특성 및 보관 정보

유전자명
NDUFB9
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.

The protein NDUFB9 also known as NADH:ubiquinone oxidoreductase subunit B9 is a part of the mitochondrial respiratory chain. This protein has a molecular mass of approximately 22 kDa. NDUFB9 is expressed in the mitochondria of many tissues serving as an accessory subunit of complex I the first enzyme in the electron transport chain. It does not participate in catalysis but assists in maintaining the integrity and stability of the complex.
Biological function summary

NDUFB9 plays a role in mitochondrial energy production. As a component of complex I which is the largest of the five mitochondrial complexes NDUFB9 contributes to the process of oxidative phosphorylation. This complex transfers electrons from NADH to ubiquinone facilitating the pumping of protons across the inner mitochondrial membrane. This proton gradient drives ATP production vital for energy metabolism in cells.

Pathways

The function of NDUFB9 is critical in cellular respiration and energy production pathways. It is integral to the pathway of oxidative phosphorylation closely associated with aerobic energy metabolism. Through its role in complex I NDUFB9 interacts with other mitochondrial proteins such as NDUFA1 and NDUFS1 which are also part of the same complex and contribute to electron transport and energy conservation.

NDUFB9 mutations have been linked to mitochondrial disorders including Leigh syndrome a severe neurological disorder characterized by progressive loss of mental and movement abilities. NDUFB9 dysfunction can also contribute to Complex I deficiency which leads to a range of metabolic syndromes. This protein's interaction with other complex I proteins like NDUFS2 is important for maintaining proper mitochondrial function and disturbances can lead to broader mitochondrial pathologies.

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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