AB264697

Human RBPMS knockout HeLa cell line

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RBPMS KO cell line available to order. KO validated. Free of charge wild type control available. Knockout achieved by using CRISPR/Cas9, 1 bp deletion in exon 2 and 44 bp deletion 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'.

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Lab

Western blot - Human RBPMS knockout HeLa cell line (AB264697)

Lanes 1 - 4 : Merged signal (red and green). Green - ab194213 observed at 30 kDa. Red - loading control ab7291 (Mouse anti-Alpha Tubulin [DM1A]) observed at 55 kDa.

ab194213 was shown to react with RBPMS in wild-type HeLa cells in western blot. Here no band was observed in the RBPMS knockout cell line ab264697 (RBPMS knockout cell lysate ab258631) but other antibodies have shown bands in this lysate below 30 kDa that may represent truncated forms and cleaved fragments. This has not been investigated further. HeLa wild-type and RBPMS knockout cell lysates were subjected to SDS-PAGE. Membranes were blocked in 3 % milk in TBS-T (0.1 % Tween^®) before incubation with ab194213 and ab7291 (Mouse anti-Alpha Tubulin [DM1A]) overnight at 4° at a 1 in 1000 dilution and a 1 in 20000 dilution respectively. Blots were incubated with Goat anti-Rabbit IgG H&L (IRDye^® 800CW) preabsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye^® 680RD) preabsorbed (ab216776) secondary antibodies at 1 in 20000 dilution for 1 h at room temperature before imaging.

Lanes 1 - 4:

Western blot - Anti-RBPMS antibody (<a href='/ko/products/primary-antibodies/rbpms-antibody-ab194213'>ab194213</a>) at 1/1000 dilution

Lanes 1 - 4:

Western blot - Anti-RBPMS antibody (<a href='/ko/products/primary-antibodies/rbpms-antibody-ab152101'>ab152101</a>) at 1/500 dilution

Lane 1:

Wild-type HeLa cell lysate at 20 µg

Lane 2:

RBPMS Knockout HeLa cell lysate at 20 µg

Lane 2:

Western blot - Human RBPMS knockout HeLa cell line (ab264697)

Lane 3:

RT-4 cell lysate at 20 µg

Lane 4:

U-251 MG cell lysate at 20 µg

Predicted band size: 22 kDa

Observed band size: 30 kDa

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Sanger Sequencing - Human RBPMS knockout HeLa cell line (AB264697)

Sanger seq

Unknown

Sanger Sequencing - Human RBPMS knockout HeLa cell line (AB264697)

Allele-1 : 1 bp deletion in exon 2.

Sanger seq

Unknown

Sanger Sequencing - Human RBPMS knockout HeLa cell line (AB264697)

Allele-2 : 44 bp deletion in exon 2.

주요 정보

세포 유형

HeLa

Species or organism

Human

조직

Cervix

제형

Liquid

form

Knockout 검증

Sanger Sequencing

Mutation 정보

Knockout achieved by using CRISPR/Cas9, 1 bp deletion in exon 2 and 44 bp deletion in exon 2

질병

Adenocarcinoma

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Reactivity 정보

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

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제품 세부 정보

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 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.

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제품 구성

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특성 및 보관 정보

유전자명

RBPMS

유전자 편집 종류

Knockout

유전자 편집 방식

CRISPR technology

Knockout 검증

Sanger Sequencing

배송 시 보관 조건

Dry Ice

적절한 단기 보관 조건

-196°C

적절한 장기 보관 조건

-196°C

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추가 정보

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

RBPMS also known as RNA Binding Protein with Multiple Splicing is an important component in RNA processing. It weighs approximately 24 kDa and is widely expressed in human tissues with significant presence in the central nervous system and heart. The protein engages directly with RNA molecules impacting their stability and translation. Its structure allows it to bind specifically to RNA influencing the fate of transcripts by modulating their processing and transport.

Biological function summary

RBPMS plays a role in the regulation of gene expression by interacting with other proteins to form ribonucleoprotein complexes. These complexes are important for the post-transcriptional management of gene expression impacting mRNA splicing and localization. RBPMS also exhibits involvement in the development of neuronal cells where it contributes to neuronal differentiation and growth by aiding in the precise processing of specific mRNA transcripts that are essential for neural functions.

Pathways

RBPMS participates in the regulation of the mRNA splicing and transport pathways. It interacts with the spliceosome complex influencing alternative splicing events that generate diverse protein isoforms. RBPMS is associated with proteins such as SMN1 which also plays a part in mRNA processing and is integral to the spliceosomal machinery. These interactions make RBPMS a pivotal mediator in translating genetic information into functional proteins therefore affecting cellular growth and differentiation.

Mutations or altered expression of RBPMS have links to neurodegenerative diseases like amyotrophic lateral sclerosis (ALS) and cardiovascular disorders. In ALS RBPMS along with proteins like TDP-43 becomes misregulated contributing to disease pathogenesis by disrupting normal RNA processing and splicing patterns. In the heart compromised expression levels of RBPMS can lead to aberrant cardiac function where it might associate with proteins such as Troponin T implicating it in heart muscle contraction regulation essential for maintaining cardiac health.

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품질 관리

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

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. 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.

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.

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제품 프로토콜

Visit the General protocols
Visit the Troubleshooting

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Product promise

저희는 고품질 시약으로 고객님의 연구를 서포트하고자 최선을 다하고 있으며, 모든 단계에서 함께 하겠습니다. 만약 제품이 기대한 성능을 보이지 않을 경우에도 Product Promise로 보호받으실 수 있습니다.

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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

Human RBPMS knockout HeLa cell line

Western blot - Human RBPMS knockout HeLa cell line (AB264697)

Sanger Sequencing - Human RBPMS knockout HeLa cell line (AB264697)

Sanger Sequencing - Human RBPMS knockout HeLa cell line (AB264697)

주요 정보

세포 유형

Species or organism

조직

제형

Knockout 검증

Mutation 정보

질병

보완 제품

Cell Lines & Lysates

Cell Lines & Lysates

Cell Lines & Lysates

Primary Antibodies

Reactivity 정보

제품 세부 정보

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

Biological function summary

Pathways

품질 관리

STR analysis

Cell culture

Biosafety level

Adherent/suspension

Gender

초기 취급 지침

Subculture 가이드라인

배양 배지

동결보존 배지

제품 프로토콜

Product promise