AB256897

Human EEA1 knockout HeLa cell lysate

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EEA1 KO cell lysate available now. KO validated. Free of charge wild type control included. Knockout achieved by using CRISPR/Cas9, 1 bp insertion in exon13.

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ZFYVE2, Endosome-associated protein p162, Early endosome antigen 1, EEA1, Zinc finger FYVE domain-containing protein 2

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Lab

Western blot - Human EEA1 knockout HeLa cell lysate (AB256897)

Lane 1 : Wild-type HeLa cell lysate (20µg)

Lane 2 : EEA1 knockout HeLa cell lysate (20µg)

Lane 3 : Daudi cell lysate (20µg)

Lane 4 : SH-SY5Y cell lysate (20µg)

Lanes 1- 4 : Merged signal (red and green). Green - ab109110 observed at 175 kDa. Red - loading control, ab8245 observed at 37 kDa.

ab109110 Anti-EEA1 antibody [EPR4245] - Early Endosome Marker was shown to specifically react with EEA1 in wild-type HeLa cells in western blot. The band observed in the knockout cell line ab261822 (knockout cell lysate ab256897) lane below 175kDa may represent truncated forms and cleaved fragments. This has not been investigated further. Wild-type and EEA1 knockout samples were subjected to SDS-PAGE. Membrane was blocked for 1 hour at room temperature in 0.1% TBST with 3% non-fat dried milk. ab109110 and Anti-GAPDH antibody [6C5] - Loading Control (ab8245) were incubated overnight at 4^°C at 1 in 10000 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-EEA1 antibody [EPR4245] - Early Endosome Marker (<a href='/ko/products/primary-antibodies/eea1-antibody-epr4245-early-endosome-marker-ab109110'>ab109110</a>) at 1/1000 dilution

Lane 1:

Wild-type HeLa cell lysate at 20 µg

Lane 2:

EEA1 knockout HeLa cell lysate at 20 µg

Lane 3:

Daudi cell lysate at 20 µg

Lane 4:

SH-SY5Y cell lysate at 20 µg

Predicted band size: 162 kDa

Observed band size: 175 kDa

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Sanger Sequencing - Human EEA1 knockout HeLa cell lysate (AB256897)

Sanger seq

Unknown

Sanger Sequencing - Human EEA1 knockout HeLa cell lysate (AB256897)

Homozygous : 1 bp insertion in exon13

주요 정보

세포 유형

HeLa

Species or organism

Human

조직

Cervix

Knockout 검증

Sanger Sequencing

Mutation 정보

Knockout achieved by using CRISPR/Cas9, 1 bp insertion in exon13.

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":"<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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제품 세부 정보

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

REACH authorisation
Abcam has not and does not intend to apply for the REACH Authorisation of customers' uses of products that contain European Authorisation list (Annex XIV) substances.
It is the responsibility of our customers to check the necessity of application of REACH Authorisation, and any other relevant authorisations, for their intended uses.

Lysate preparation: Our lysates are made using RIPA buffer to which we add a protease inhibitor cocktail and phosphatase inhibitor cocktail (ratio: 300:100:10). This means that the protein of interest is denatured. If you require a native form of the protein please use the live cell version. Please refer to our lysis protocol for further details on how our lysates are prepared.

User storage instructions: Lyophilizate may be stored at 4°C. After reconstitution, store at -20°C for short-term storage or -80°C for long-term storage.

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": { "1Kit": { "sellingSize": "1 Kit", "publicAssetCode":"ab256897-1Kit", "assetComponentDetails": [ { "size":"1 x 100 µg", "name":"Human EEA1 knockout HeLa cell lysate", "number":"AB256897-CMP01", "productcode":"" }, { "size":"1 x 100 µg", "name":"Human wild-type HeLa cell lysate", "number":"AB256897-CMP02", "productcode":"" } ] } } }

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특성 및 보관 정보

유전자명

EEA1

유전자 편집 종류

Knockout

유전자 편집 방식

CRISPR technology

Knockout 검증

Sanger Sequencing

배송 시 보관 조건

Ambient - Can Ship with Ice

적절한 단기 보관 조건

-20°C

적절한 장기 보관 조건

-20°C

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추가 정보

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

EEA1 (Early Endosome Antigen 1) is an important protein involved in endosomal trafficking with a molecular weight of approximately 162 kDa. It is a cytoplasmic protein expressed in various cell types acting mainly as an endosome marker. This protein contains a FYVE domain which facilitates binding to phosphatidylinositol 3-phosphate on endosomal membranes. EEA1 also interacts with Rab5 a small GTPase contributing to the fusion of early endosomes.

Biological function summary

EEA1 plays a significant role in early endosomal pathways facilitating endocytic vesicle fusion. EEA1 does not usually work alone; it functions as part of a larger complex involving other endosomal proteins. This protein acts as a tethering molecule which stabilizes the interaction of vesicles with endosomes. By anchoring vesicles EEA1 ensures the correct delivery and sorting of cargo within the cell which is important for cellular homeostasis and signaling.

Pathways

EEA1 contributes to the endocytic pathway and receptor recycling. It ensures proper functioning of the endocytic recycling of receptors such as the epidermal growth factor receptor (EGFR). EEA1-mediated tethering is vital for these pathways and it works closely with proteins like Rab5 which regulate endocytic trafficking. Through these interactions EEA1 affects cellular processes by controlling the flow of signaling receptors and other molecules within the cell.

EEA1 dysfunction is linked to neurodegenerative diseases and cancer. Abnormalities in its expression or function can disrupt endosomal trafficking leading to cellular stress and disease. For example in Alzheimer's disease impaired endosomal function involving EEA1 can contribute to amyloid-beta accumulation. In cancer altered EEA1 activity may contribute to aberrant receptor signaling which can promote unchecked cell division. The interplay with proteins like EGFR demonstrates EEA1's importance in maintaining cellular health and response mechanisms.

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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

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제품 프로토콜

Visit the General protocols
Visit the Troubleshooting

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Product promise

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Please note: All products are 'FOR RESEARCH USE ONLY. NOT FOR USE IN DIAGNOSTIC OR THERAPEUTIC PROCEDURES'.

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Human EEA1 knockout HeLa cell lysate

Western blot - Human EEA1 knockout HeLa cell lysate (AB256897)

Sanger Sequencing - Human EEA1 knockout HeLa cell lysate (AB256897)

주요 정보

세포 유형

Species or organism

조직

Knockout 검증

Mutation 정보

질병

보완 제품

Primary Antibodies

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

제품 프로토콜

Product promise