AB266738

Human GRN (Granulin) knockout HEK-293T cell line

Advanced Validation
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GRN KO cell line available to order. KO validated by Western blot. Free of charge wild type control available. Knockout achieved by using CRISPR/Cas9, 14 bp deletion in exon 2 and 2 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'.

5 이미지

Lab

Western blot - Human GRN (Granulin) knockout HEK-293T cell line (AB266738)

Lanes 1-4 : Merged signal (red and green). Green - ab208777 observed at 74 kDa. Red - loading control ab8245 observed at 36 kDa.

ab208777 Anti-Progranulin antibody [EPR15864] was shown to specifically react with Progranulin in wild-type HEK293T cells. Loss of signal was observed when knockout cell line ab266738 (knockout cell lysate ab257235) was used. Wild-type and Granulin knockout samples were subjected to SDS-PAGE. ab208777 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-Progranulin antibody [EPR15864] (<a href='/ko/products/primary-antibodies/granulin-antibody-epr15864-ab208777'>ab208777</a>) at 1/1000 dilution

Lane 1:

Wild-type HEK293T cell lysate at 20 µg

Lane 2:

GRN knockout HEK293T cell lysate at 20 µg

Lane 2:

Western blot - Human GRN (Granulin) knockout HEK-293T cell line (ab266738)

Lane 3:

HepG2 cell lysate at 20 µg

Lane 4:

HAP1 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: 64 kDa

Observed band size: 74 kDa

false

Sanger Sequencing - Human GRN (Granulin) knockout HEK-293T cell line (AB266738)

Sanger seq

Unknown

Sanger Sequencing - Human GRN (Granulin) knockout HEK-293T cell line (AB266738)

Allele-1 : 14 bp deletion in exon2

Cell Culture - Human GRN (Granulin) knockout HEK-293T cell line (AB266738)

Cell Culture

Unknown

Cell Culture - Human GRN (Granulin) knockout HEK-293T cell line (AB266738)

Representative images of GRN knockout HEK293T cells, low and high confluency examples (top left and right respectively) and wild-type HEK293T cells, low and high confluency (bottom left and right respectively) showing typical adherent, epithelial-like morphology. Images were captured at 10X magnification using an EVOS M5000 microscope.

Sanger seq

Unknown

Sanger Sequencing - Human GRN (Granulin) knockout HEK-293T cell line (AB266738)

Allele-2 : 2 bp deletion in exon 2.

Lab

Western blot - Human GRN (Granulin) knockout HEK-293T cell line (AB266738)

All lanes:

Western blot - Anti-Progranulin antibody [EPR15864] (<a href='/ko/products/primary-antibodies/granulin-antibody-epr15864-ab208777'>ab208777</a>) at 1/1000 dilution

Lane 1:

Wild-type HEK293T lysate at 20 µg

Lane 2:

Western blot - Human GRN (Granulin) knockout HEK-293T cell line (ab266738)

Lane 2:

Western blot - Human GRN (Granulin) knockout HEK-293T cell lysate (<a href='/ko/products/cell-lysates/human-grn-granulin-knockout-hek-293t-cell-lysate-ab257235'>ab257235</a>)

Observed band size: 64 kDa

false

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, 14 bp deletion in exon 2 and 2 bp deletion in exon 2

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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

{ "values": { "2x1000000Cellsvial": { "sellingSize": "2 x 1000000 Cells/vial", "publicAssetCode":"ab266738-2x1000000Cells_vial", "assetComponentDetails": [ { "size":"1 x 1000000 Cells/vial", "name":"ab266738 Human GRN (Granulin) knockout HEK-293T cell line", "number":"AB266738-CMP01" }, { "size":"1 x 1000000 Cells/vial", "name":"ab255449 Human wild-type HEK-293T cell line", "number":"AB266738-CMP02" } ] }, "1000000Cellsvial": { "sellingSize": "1000000 Cells/vial", "publicAssetCode":"ab266738-1000000Cells_vial", "assetComponentDetails": [ { "size":"1 x 1000000 Cells/vial", "name":"ab266738 Human GRN (Granulin) knockout HEK-293T cell line", "number":"AB266738-CMP01", "productcode":"" } ] } } }

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특성 및 보관 정보

유전자명

GRN

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.

Granulin also known as progranulin or GRN is a protein with a molecular mass of approximately 68 kDa. This protein is encoded by the GRN gene and is involved in various cellular processes. Granulin is expressed widely within the body particularly in epithelial cells immune cells and neurons. Researchers have developed methods like GRN ELISA and progranulin ELISA to quantitatively measure the levels of granulin in biological samples aiding in further understanding of its role in health and disease.

Biological function summary

Granulin functions as a growth factor involved in tissue repair cell proliferation and inflammation. It does not function as part of a larger protein complex but acts individually in exerting its effects on cells. Granulin influences various aspects of cellular architecture and signaling pathways making it important for maintaining cellular homeostasis. Additionally the granulin protein participates in regulating networks that involve cytokines and growth factors impacting both the innate and adaptive immune responses.

Pathways

Granulin plays significant roles in the MAPK and NF-kB signaling pathways. These pathways critically manage cellular reactions to stress inflammation and other external stimuli. Granulin modulates these processes often interacting with proteins like TNF receptors and interleukins to affect the downstream signaling events. Interactions between granulin and other signaling molecules allow it to integrate into broader networks that oversee various biological responses.

Significant links exist between granulin mutations and frontotemporal dementia (FTD) and Alzheimer's disease. Mutations in the GRN gene often lead to reduced granulin levels contributing to neurodegenerative conditions. These mutations affect the protein's normal function promoting pathological mechanisms associated with neuronal deterioration. Granulin interacts with proteins such as TDP-43 in neurodegenerative diseases where alterations might result in protein aggregation and neurotoxicity. Understanding these connections can lead to potential therapeutic interventions targeting granulin-related pathways.

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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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타겟 정보

See full target information GRN

대체 명칭 보기

PEPI, PC cell-derived growth factor, Acrogranin, Granulin precursor, GP88, GRN, PCDGF, Glycoprotein of 88 Kda, Proepithelin, Glycoprotein 88, Epithelin precursor, Progranulin, PGRN

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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 GRN (Granulin) knockout HEK-293T cell line

Western blot - Human GRN (Granulin) knockout HEK-293T cell line (AB266738)

Sanger Sequencing - Human GRN (Granulin) knockout HEK-293T cell line (AB266738)

Cell Culture - Human GRN (Granulin) knockout HEK-293T cell line (AB266738)

Sanger Sequencing - Human GRN (Granulin) knockout HEK-293T cell line (AB266738)

Western blot - Human GRN (Granulin) knockout HEK-293T cell line (AB266738)

주요 정보

세포 유형

Species or organism

조직

제형

Knockout 검증

Mutation 정보

보완 제품

Cell Lines & Lysates

Primary Antibodies

Primary Antibodies

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

타겟 정보

대체 명칭 보기

Product promise