AB257280

Human SLC9A3R1 (EBP50/NHERF-1) knockout HeLa cell lysate

Name: Human SLC9A3R1 (EBP50/NHERF-1) knockout HeLa cell lysate (ab257280) | Abcam
Brand: Abcam Limited
SKU: AB257280
Price: 111000 JPY
Availability: InStock

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NHERF1 KO cell lysate available now. KO validated by Western blot. Free of charge wild type control included. Knockout achieved by using CRISPR/Cas9, Homozygous: 19 bp deletion in exon 1.

3 Images

Lab

Western blot - Human SLC9A3R1 (EBP50/NHERF-1) knockout HeLa cell lysate (AB257280)

False colour image of Western blot : Anti-EBP50/NHERF-1 antibody [EPR5562] staining at 1/1000 dilution shown in green; Mouse anti-Alpha Tubulin [DM1A] (ab7291) loading control staining at 1/20000 dilution shown in red. In Western blot ab109430 was shown to bind specifically to EBP50/NHERF-1. A band was observed at 46 kDa in wild-type HeLa cell lysates with no signal observed at this size in SLC9A3R1 knockout cell line ab264914 (knockout cell lysate ab257280). To generate this image wild-type and SLC9A3R1 knockout HeLa cell lysates were analysed. First samples were run on an SDS-PAGE gel then transferred onto a nitrocellulose membrane. Membranes were blocked in fluorescent western blot (TBS-based) blocking solution before incubation with primary antibodies overnight at 4°. Blots were washed four times in TBS-T incubated with secondary antibodies for 1 h at room temperature washed again four times then imaged.Secondary antibodies used were Goat anti-Rabbit IgG H&L (IRDye^® 800CW) preabsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye^® 680RD) preabsorbed (ab216776) at 1/20000 dilution.

All lanes:

Western blot - Anti-EBP50/NHERF-1 antibody [EPR5562] (<a href='/products/primary-antibodies/ebp50-nherf-1-antibody-epr5562-ab109430'>ab109430</a>) at 1/1000 dilution

Lane 1:

Wild-type HeLa cell lysate at 20 µg

Lane 2:

SLC9A3R1 knockout HeLa cell lysate at 20 µg

Lane 2:

Western blot - Human SLC9A3R1 (EBP50/NHERF-1) knockout HeLa cell line (<a href='/products/cell-lines/human-slc9a3r1-ebp50-nherf-1-knockout-hela-cell-line-ab264914'>ab264914</a>)

Lane 2:

Western blot - Human SLC9A3R1 (EBP50/NHERF-1) knockout HeLa cell lysate (ab257280)

Predicted band size: 38 kDa

Observed band size: 46 kDa

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Lab

Western blot - Human SLC9A3R1 (EBP50/NHERF-1) knockout HeLa cell lysate (AB257280)

False colour image of Western blot : Anti-EBP50/NHERF-1 antibody staining at 1 μg/ml shown in green; Mouse anti-Alpha Tubulin [DM1A] (ab7291) loading control staining at 1/20000 dilution shown in red. In Western blot ab88238 was shown to bind specifically to EBP50/NHERF-1. A band was observed at 46 kDa in wild-type HeLa cell lysates with no signal observed at this size in SLC9A3R1 knockout cell line ab264914 (knockout cell lysate ab257280). To generate this image wild-type and SLC9A3R1 knockout HeLa cell lysates were analysed. First samples were run on an SDS-PAGE gel then transferred onto a nitrocellulose membrane. Membranes were blocked in fluorescent western blot (TBS-based) blocking solution before incubation with primary antibodies overnight at 4 °. Blots were washed four times in TBS-T incubated with secondary antibodies for 1 h at room temperature washed again four times then imaged.Secondary antibodies used were Goat anti-Rabbit IgG H&L (IRDye^® 800CW) preabsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye^® 680RD) preabsorbed (ab216776) at 1/20000 dilution.

All lanes:

Western blot - Anti-EBP50/NHERF-1 antibody (<a href='/products/primary-antibodies/ebp50-nherf-1-antibody-ab88238'>ab88238</a>) at 1 µg/mL

Lane 1:

Wild-type HeLa cell lysate at 20 µg

Lane 2:

SLC9A3R1 knockout HeLa cell lysate at 20 µg

Lane 2:

Western blot - Human SLC9A3R1 (EBP50/NHERF-1) knockout HeLa cell line (<a href='/products/cell-lines/human-slc9a3r1-ebp50-nherf-1-knockout-hela-cell-line-ab264914'>ab264914</a>)

Lane 2:

Western blot - Human SLC9A3R1 (EBP50/NHERF-1) knockout HeLa cell lysate (ab257280)

Predicted band size: 38 kDa

Observed band size: 46 kDa

false

Sanger Sequencing - Human SLC9A3R1 (EBP50/NHERF-1) knockout HeLa cell lysate (AB257280)

Sanger seq

Unknown

Sanger Sequencing - Human SLC9A3R1 (EBP50/NHERF-1) knockout HeLa cell lysate (AB257280)

Homozygous : 19 bp deletion in exon 1

Key facts

細胞タイプ

HeLa

生物種

Human

組織

Cervix

ノックアウト検証方法

Sanger Sequencing,Western blot

ノックアウト変異

Knockout achieved by using CRISPR/Cas9, Homozygous: 19 bp deletion in exon 1.

疾病

Adenocarcinoma

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Reactivity data

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

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製品の詳細

Knockout cell lysate achieved by CRISPR/Cas9.

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 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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製品内容

{ "values": { "1Kit": { "sellingSize": "1 Kit", "publicAssetCode":"ab257280-1Kit", "assetComponentDetails": [ { "size":"1 x 100 µg", "name":"Human SLC9A3R1 knockout HeLa cell lysate", "number":"AB257280-CMP02", "productcode":"" }, { "size":"1 x 100 µg", "name":"Human wild-type HeLa cell lysate", "number":"AB257280-CMP01", "productcode":"" } ] } } }

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出荷温度及び保存条件

遺伝子名

NHERF1

遺伝子編集のタイプ

Knockout

遺伝子編集の方法

CRISPR technology

ノックアウト検証方法

Sanger Sequencing, Western blot

接合型

Homozygous

出荷温度

Ambient - Can Ship with Ice

短期保存温度

-20°C

長期保存温度

-20°C

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補足情報

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

EBP50 also known as NHERF-1 (Na+/H+ exchanger regulatory factor 1) is a scaffolding protein with a mass of approximately 50 kDa. This protein plays a critical role in the formation of multiprotein complexes by its ability to bind to several different proteins simultaneously. EBP50 localizes primarily to the apical surface of epithelial cells where it is found in organs such as the kidney liver and intestines. It consists of two PDZ domains and an ezrin-binding domain which enable it to interact with various membrane proteins cytoskeletal elements and signaling molecules.

Biological function summary

EBP50 regulates cellular functions by organizing signaling complexes at the plasma membrane. It is involved in processes such as ion transport signal transduction and cell proliferation. EBP50 often acts as part of a multiprotein complex contributing to the stabilization and regulation of membrane receptors and channels. By regulating these components EBP50 affects a variety of cellular responses to external stimuli.

Pathways

EBP50 plays important roles in cellular signaling pathways like the PTH (parathyroid hormone) receptor signaling and CFTR (cystic fibrosis transmembrane conductance regulator) channel regulation. It interacts with proteins such as ezrin and PLCβ (phospholipase C beta) which facilitates the signaling cascades critical for maintaining proper cell function. By modulating these pathways EBP50 has a significant impact on maintaining cellular homeostasis and responding to changing physiological conditions.

EBP50 has been linked to conditions such as cystic fibrosis and certain cancers. With cystic fibrosis its interaction with CFTR is essential for the proper regulation of chloride channels and subsequently fluid balance in epithelial tissues. In cancer altered expression or mutations in EBP50 can affect pathways related to cell growth and metastasis highlighting interactions with the ezrin protein which couples signals from the cell membrane to actin cytoskeleton dynamics impacting tumor progression.

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Quality control

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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製品プロトコール

Visit the General protocols
Visit the Troubleshooting

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Abcam product promise

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Human SLC9A3R1 (EBP50/NHERF-1) knockout HeLa cell lysate