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AB261824

Human EIF2AK2 (PKR) knockout HeLa cell line

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EIF2AK2 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 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 EIF2AK2 (PKR) knockout HeLa cell line (AB261824)
  • WB

Lab

Western blot - Human EIF2AK2 (PKR) knockout HeLa cell line (AB261824)

Lanes 1-5 : Merged signal (red and green). Green - ab32052 observed at 70 kDa. Red - loading control ab8245 observed at 36 kDa.

ab32052 Anti-PKR antibody [YE350] was shown to specifically react with PKR in wild-type HeLa cells. Loss of signal was observed when knockout cell line ab261824 (knockout cell lysate ab256899) was used. Wild-type and PKR knockout samples were subjected to SDS-PAGE. ab32052 and Anti-GAPDH antibody [6C5] - Loading Control (ab8245) were incubated overnight at 4°C 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-PKR antibody [YE350] (<a href='/ko/products/primary-antibodies/pkr-antibody-ye350-ab32052'>ab32052</a>) at 1/1000 dilution

Lane 1:

Wild-type HeLa (Human epithelial cell line from cervix adenocarcinoma) whole cell lysate at 20 µg

Lane 2:

EIF2AK2 knockout HeLa (Human epithelial cell line from cervix adenocarcinoma) whole cell lysate at 20 µg

Lane 2:

Western blot - Human EIF2AK2 (PKR) knockout HeLa cell lysate (<a href='/ko/products/cell-lysates/human-eif2ak2-pkr-knockout-hela-cell-lysate-ab256899'>ab256899</a>) at 20 µg

Lane 3:

Wild-type A549 (Human lung carcinoma cell line) whole cell lysate at 20 µg

Lane 4:

EIF2AK2 knockout A549 (Human lung carcinoma cell line) whole cell lysate at 20 µg

Lane 5:

K562 (Human chronic myelogenous leukemia lymphoblast cell line) whole 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: 62 kDa

Observed band size: 70 kDa

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Western blot - Human EIF2AK2 (PKR) knockout HeLa cell line (AB261824)
  • WB

Lab

Western blot - Human EIF2AK2 (PKR) knockout HeLa cell line (AB261824)

Lanes 1-5 : Merged signal (red and green). Green - ab184257 observed at 70 kDa. Red - loading control ab8245 observed at 36 kDa.

ab184257 Anti-PKR antibody [EPR19374] was shown to specifically react with PKR in wild-type HeLa cells. Loss of signal was observed when knockout cell line ab261824 (knockout cell lysate ab256899) was used. Wild-type and PKR knockout samples were subjected to SDS-PAGE. ab184257 and Anti-GAPDH antibody [6C5] - Loading Control (ab8245) were incubated overnight at 4°C 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-PKR antibody [EPR19374] (<a href='/ko/products/primary-antibodies/pkr-antibody-epr19374-ab184257'>ab184257</a>) at 1/1000 dilution

Lane 1:

Wild-type HeLa (Human epithelial cell line from cervix adenocarcinoma) whole cell lysate at 20 µg

Lane 2:

EIF2AK2 knockout HeLa (Human epithelial cell line from cervix adenocarcinoma) whole cell lysate at 20 µg

Lane 2:

Western blot - Human EIF2AK2 (PKR) knockout HeLa cell line (ab261824)

Lane 3:

Wild-type A549 (Human lung carcinoma cell line) whole cell lysate at 20 µg

Lane 4:

EIF2AK2 knockout A549 (Human lung carcinoma cell line) whole cell lysate at 20 µg

Lane 5:

K562 (Human chronic myelogenous leukemia lymphoblast cell line) whole 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: 62 kDa

Observed band size: 70 kDa

false

Cell Culture - Human EIF2AK2 (PKR) knockout HeLa cell line (AB261824)
  • Cell Culture

Lab

Cell Culture - Human EIF2AK2 (PKR) knockout HeLa cell line (AB261824)

Representative images EIF2AK2 knockout HeLa cells, low and high confluency examples (top left and right respectively) and wild-type HeLa cells, low and high confluency (bottom left and right respectively) showing typical adherent, epithelial-like morphology. Images were captured at 10X magnification using a EVOS M5000 microscope.

Sanger Sequencing - Human EIF2AK2 (PKR) knockout HeLa cell line (AB261824)
  • Sanger seq

Unknown

Sanger Sequencing - Human EIF2AK2 (PKR) knockout HeLa cell line (AB261824)

Homozygous : 1 bp insertion in exon 1.

Sanger Sequencing - Human EIF2AK2 (PKR) knockout HeLa cell line (AB261824)
  • Sanger seq

Lab

Sanger Sequencing - Human EIF2AK2 (PKR) knockout HeLa cell line (AB261824)

Sequencing chromatogram displaying sequence edit in exon 1

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 1

Antibiotic resistance

Puromycin 1µg/mL

질병

Adenocarcinoma

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보완 제품

Cell Lines & Lysates

AB266999

Human EIF2AK2 (PKR) knockout A549 cell line

Advanced Validation

Western blot - Human EIF2AK2 (PKR) knockout A549 cell line (AB266999)

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Cell Lines & Lysates

AB267108

Human OAS1 knockout A549 cell line

Advanced Validation

Western blot - Human OAS1 knockout A549 cell line (AB267108)

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Proteins & Peptides

AB93699

Recombinant human Lactate Dehydrogenase protein (Active) (His tag N-Terminus)

Western blot - Recombinant human Lactate Dehydrogenase protein (Active) (His tag N-Terminus) (AB93699)

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Reactivity 정보

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

{ "values": { "2x1000000Cellsvial": { "sellingSize": "2 x 1000000 Cells/vial", "publicAssetCode":"ab261824-2x1000000Cells_vial", "assetComponentDetails": [ { "size":"1 x 1000000 Cells/vial", "name":"ab255928 Human wild-type HeLa cell line", "number":"AB261824-CMP02" }, { "size":"1 x 1000000 Cells/vial", "name":"ab261824 Human EIF2AK2 (PKR) knockout HeLa cell line", "number":"AB261824-CMP01" } ] }, "1000000Cellsvial": { "sellingSize": "1000000 Cells/vial", "publicAssetCode":"ab261824-1000000Cells_vial", "assetComponentDetails": [ { "size":"1 x 1000000 Cells/vial", "name":"ab261824 Human EIF2AK2 (PKR) knockout HeLa cell line", "number":"AB261824-CMP01", "productcode":"" } ] } } }
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특성 및 보관 정보

유전자명
EIF2AK2
유전자 편집 종류
Knockout
유전자 편집 방식
CRISPR technology
Knockout 검증
Sanger Sequencing, Western blot
Zygosity
Homozygous
품질 관리 - STR analysis
CSF1PO, D13S317, D7S820, D5S818, TH01, D16S539, TPOX
배송 시 보관 조건
Dry Ice
적절한 단기 보관 조건
-196°C
적절한 장기 보관 조건
-196°C
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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. <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.

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제품 프로토콜

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타겟 정보

See full target information EIF2AK2

대체 명칭 보기

P1/eIF-2A protein kinase, EIF2AK2, Protein kinase RNA-activated, Protein kinase R, eIF-2A protein kinase 2, Eukaryotic translation initiation factor 2-alpha kinase 2, p68 kinase, Tyrosine-protein kinase EIF2AK2, PKR, Interferon-inducible RNA-dependent protein kinase, PRKR
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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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