AB264845

Human RNF2 (RING2 / RING1B) knockout HeLa cell line

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RNF2 KO cell line available to order. KO validated. Free of charge wild type control available. Knockout achieved by using CRISPR/Cas9, 8 bp deletion in exon 2 and Insertion of the selection cassette 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'.

別名を表示する

DING, DinG protein, E3 ubiquitin protein-ligase RING 2, E3 ubiquitin-protein ligase RING2, HIP2-interacting protein 3, HIPI 3, Huntingtin-interacting protein 2-interacting protein 3, OTTHUMP00000060668, Polycomb M33 interacting protein Ring 1B, Protein DinG, RING 1B, RING finger protein 1B, RING finger protein 2, RING finger protein BAP-1, RING2_HUMAN, RNF 2

3 Images

Lab

Western blot - Human RNF2 (RING2 / RING1B) knockout HeLa cell line (AB264845)

Lanes 1- 2 : Merged signal (red and green). Green - ab181140 observed at 42 kDa. Red - Anti-alpha Tubulin antibody [DM1A] - Loading Control (ab7291) observed at 50 kDa.

ab181140 was shown to react with RING2 / RING1B / RNF2 in wild-type HeLa cells in western blot. The band observed in knockout cell line ab264845 (knockout cell lysate ab257640) lane below 42kDa may represent truncated forms and cleaved fragments. This has not been investigated further. Wild-type HeLa and RNF2 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. ab181140 and Anti-alpha Tubulin antibody [DM1A] - Loading Control (ab7291) were incubated 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-RING2 / RING1B / RNF2 antibody [EPR12245] (<a href='/products/primary-antibodies/ring2-ring1b-rnf2-antibody-epr12245-ab181140'>ab181140</a>) at 1/1000 dilution

Lane 1:

Wild-type HeLa cell lysate at 20 µg

Lane 2:

RNF2 CRISPR/Cas9 edited HeLa cell lysate at 20 µg

Lane 2:

Western blot - Human RNF2 (RING2 / RING1B) knockout HeLa cell line (ab264845)

Predicted band size: 37 kDa

Observed band size: 42 kDa

false

Sanger Sequencing - Human RNF2 (RING2 / RING1B) knockout HeLa cell line (AB264845)

Sanger seq

Unknown

Sanger Sequencing - Human RNF2 (RING2 / RING1B) knockout HeLa cell line (AB264845)

Allele-1 : 8 bp deletion in exon 2.

Sanger seq

Unknown

Sanger Sequencing - Human RNF2 (RING2 / RING1B) knockout HeLa cell line (AB264845)

Allele-2 : Insertion of the selection cassette in exon 2.

Key facts

細胞タイプ

HeLa

生物種

Human

組織

Cervix

製品の状態

Liquid

form

ノックアウト検証方法

Sanger Sequencing

ノックアウト変異

Knockout achieved by using CRISPR/Cas9, 8 bp deletion in exon 2 and Insertion of the selection cassette in exon 2

疾病

Adenocarcinoma

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Reactivity data

{ "title": "Reactivity Data", "filters": { "stats": ["", "Reactivity", "Dilution Info", "Notes"] }, "values": { "WB": { "reactivity":"TESTED_AND_REACTS", "dilution-info":"", "notes":"<p>Western blot data indicates that the CRISPR gene edit may have resulted in a truncation of the protein of interest. Please see data images.</p>" } } }

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製品の詳細

We will provide viable cells that proliferate on revival.

Western blot data indicates that the CRISPR gene edit may have resulted in a truncation of the protein of interest. Please see data images.

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.

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製品内容

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出荷温度及び保存条件

遺伝子名

RNF2

遺伝子編集のタイプ

Knockout

遺伝子編集の方法

CRISPR technology

ノックアウト検証方法

Sanger Sequencing

出荷温度

Dry Ice

短期保存温度

-196°C

長期保存温度

-196°C

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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

DMEM (High Glucose) + 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.

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補足情報

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

RING2 also known as RING1B or RNF2 is a protein that functions as a member of the Polycomb Repressive Complex 1 (PRC1). This protein contains a RING-finger domain facilitating ubiquitin ligase activity. It has a molecular mass of approximately 38 kDa. RING2/RNF2/RING1B is expressed in various tissues including the brain heart and lungs indicating its fundamental role across different biological systems.

Biological function summary

RING2/RNF2/RING1B regulates gene expression by modifying chromatin structure especially through monoubiquitination of histone H2A at lysine 119. This action occurs within the Polycomb Repressive Complex 1 a multiprotein structure important for maintaining the transcriptional repression of target genes. It influences cell cycle progression and development by silencing genes involved in proliferation and differentiation.

Pathways

RING2/RNF2/RING1B's role connects to the Polycomb group (PcG) pathway and the Wnt signaling pathway. Within these pathways it interacts with proteins like BMI1 and EZH2 which are other components of polycomb complexes. These interactions help modulate chromatin dynamics and control the expression of several genes involved in cell identity and development.

Dysfunction or aberrant expression of RING2/RNF2/RING1B has links to cancer and neurological disorders. Its relationship with proteins like CBX8 ties it to pathways of oncogenesis where it can influence tumorigenesis through misregulation of gene silencing. Furthermore changes in RING2/RNF2/RING1B expression or function may contribute to developmental disorders associated with improper neuronal development.

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Quality control

STR analysis

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

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Cell culture

Biosafety level

EU: 2 US: 2

Adherent/suspension

Adherent

Gender

Female

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製品プロトコール

Visit the General protocols
Visit the Troubleshooting

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ターゲットの情報

See full target information RNF2

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Human RNF2 (RING2 / RING1B) knockout HeLa cell line