AB266806

Human PRKRA (PACT) knockout HEK-293T cell line

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PRKRA KO cell line available to order. KO validated by Western blot. Free of charge wild type control available. Knockout achieved by using CRISPR/Cas9, 1 bp deletion in exon 1 and 2 bp deletion in exon 1. 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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Western blot - Human PRKRA (PACT) knockout HEK-293T cell line (AB266806)

Lanes 1-4 : Merged signal (red and green). Green - ab31967 observed at 36 kDa. Red - loading control ab8245 observed at 36 kDa.

ab31967 Anti-PACT (PKR activating protein) / PRKRA antibody was shown to specifically react with PACT in wild-type HEK-293T cells. Loss of signal was observed when knockout cell line ab266806 (knockout cell lysate ab258141) was used. Wild-type and PACT knockout samples were subjected to SDS-PAGE. ab31967 and Anti-GAPDH antibody [6C5] - Loading Control (ab8245) were incubated at room temperature for 2. 5 hours 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-PACT (PKR activating protein) / PRKRA antibody (<a href='/ko/products/primary-antibodies/pact-pkr-activating-protein-prkra-antibody-ab31967'>ab31967</a>) at 1/1000 dilution

Lane 1:

Wild-type HEK-293T cell lysate at 20 µg

Lane 2:

PRKRA knockout HEK-293T cell lysate at 20 µg

Lane 2:

Western blot - Human PRKRA (PACT) knockout HEK-293T cell line (ab266806)

Lane 3:

K-562 cell lysate at 20 µg

Lane 4:

HepG2 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: 34 kDa

Observed band size: 36 kDa

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Lab

Western blot - Human PRKRA (PACT) knockout HEK-293T cell line (AB266806)

Lanes 1-4 : Merged signal (red and green). Green - ab75749 observed at 36 kDa. Red - loading control ab8245 observed at 36 kDa.

ab75749 Anti-PACT (PKR activating protein) / PRKRA antibody [EPR3224] was shown to specifically react with PACT in wild-type HEK-293T cells. Loss of signal was observed when knockout cell line ab266806 (knockout cell lysate ab258141) was used. Wild-type and PACT knockout samples were subjected to SDS-PAGE. ab75749 and Anti-GAPDH antibody [6C5] - Loading Control (ab8245) were incubated at room temperature for 2.5 hours 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-PACT (PKR activating protein) / PRKRA antibody [EPR3224] (<a href='/ko/products/primary-antibodies/pact-pkr-activating-protein-prkra-antibody-epr3224-ab75749'>ab75749</a>) at 1/1000 dilution

Lane 1:

Wild-type HEK-293T cell lysate at 20 µg

Lane 2:

PRKRA knockout HEK-293T cell lysate at 20 µg

Lane 2:

Western blot - Human PRKRA (PACT) knockout HEK-293T cell line (ab266806)

Lane 3:

K-562 cell lysate at 20 µg

Lane 4:

HepG2 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: 34 kDa

Observed band size: 36 kDa

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Sanger Sequencing - Human PRKRA (PACT) knockout HEK-293T cell line (AB266806)

Sanger seq

Unknown

Sanger Sequencing - Human PRKRA (PACT) knockout HEK-293T cell line (AB266806)

Allele-2 : 1 bp deletion in exon 1.

Sanger seq

Unknown

Sanger Sequencing - Human PRKRA (PACT) knockout HEK-293T cell line (AB266806)

Allele-1 : 2 bp deletion in exon1

주요 정보

세포 유형

HEK-293T

Species or organism

Human

조직

Kidney

제형

Liquid

form

Knockout 검증

Sanger Sequencing,Western blot

Mutation 정보

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

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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.

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

PRKRA

유전자 편집 종류

Knockout

유전자 편집 방식

CRISPR technology

Knockout 검증

Sanger Sequencing, Western blot

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.

PACT also known as PKR activating protein or PRKRA is a regulator of the protein kinase R (PKR). This protein plays a role in PKR activation stimulating its autophosphorylation. Structurally PACT has a molecular mass of approximately 37 kDa. Researchers observed its expression in many tissues with notable presence in the brain and in immune cells. PACT functions interact with PKR and it also goes by the alternative names PKR activator and PKR protein activator.

Biological function summary

PACT involves in the cellular stress response signaling. It serves as an activator for PKR a kinase that becomes active during viral infections and stress. PACT forms complexes with PKR facilitating its activation and subsequent initiation of downstream signaling cascades. It contributes significantly to the regulation of protein synthesis and cell growth by modulating these pathways. In conditions of stress PACT ensures adequate PKR activation for host defense.

Pathways

PACT is key in the interferon signaling pathway and the stress response pathway. It partners with PKR to suppress protein synthesis through phosphorylation of the eukaryotic initiation factor 2 alpha (eIF2α) which slows down protein production during cellular stress. This interaction places PACT at a junction of antiviral defense and stress adaptation with PKR serving as an essential protein involved in these pathways.

PACT links to conditions such as Huntington's disease and dystonia. Dysregulation in PACT's function impacts these neurodegenerative disorders as it relates to its role in stress response. Altered interactions between PACT and PKR contribute to neuronal dysfunction highlighting its importance in these pathologies. Understanding how PACT and its partner proteins like PKR and eIF2α operate can lead to potential therapeutic targets for these diseases.

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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.

자세한 내용은 이용 약관을 확인해 주세요.

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 PRKRA (PACT) knockout HEK-293T cell line

Western blot - Human PRKRA (PACT) knockout HEK-293T cell line (AB266806)

Western blot - Human PRKRA (PACT) knockout HEK-293T cell line (AB266806)

Sanger Sequencing - Human PRKRA (PACT) knockout HEK-293T cell line (AB266806)

Sanger Sequencing - Human PRKRA (PACT) knockout HEK-293T cell line (AB266806)

주요 정보

세포 유형

Species or organism

조직

제형

Knockout 검증

Mutation 정보

보완 제품

Primary Antibodies

Primary Antibodies

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