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AB259109

Human SH3GL1 (EEN) knockout HEK-293T cell lysate

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SH3GL1 KO cell lysate available now. KO validated by Western blot. Free of charge wild type control included. Knockout achieved by using CRISPR/Cas9, 10 bp deletion in exon 2 and Insertion of the selection cassette in exon 2.
3 Images
Western blot - Human SH3GL1 (EEN) knockout HEK-293T cell lysate (AB259109)
  • WB

Lab

Western blot - Human SH3GL1 (EEN) knockout HEK-293T cell lysate (AB259109)

Lane 1 : Wild-type HEK-293T cell lysate 20 ug
Lane 2 : SH3GL1 knockout HEK-293T cell lysate 20 ug
Lane 3 : HUVEC cell lysate 20 ug
Lane 4 : A431 cell lysate 20 ug
Lanes 1 - 4 : Merged signal (red and green). Green - Mouse monoclonal antibody observed at 48 kDa. Red - loading control ab52866 (Rabbit anti-alpha Tubulin antibody [EP1332Y]) observed at 55kDa.
Mouse monoclonal antibody was shown to react with Endophilin II in wild-type HEK-293T cells in Western blot with loss of signal observed in SH3GL1 knockout cell line ab266260 (SH3GL1 knockout cell lysate ab259109). Wild-type HEK-293T and SH3GL1 knockout cell lysates were subjected to SDS-PAGE. Membranes were blocked in 3 % milk in TBS-T (0.1 % Tween®) before incubation with Mouse monoclonal antibody and ab52866 (Rabbit anti-alpha Tubulin antibody [EP1332Y]) overnight at 4 °C at a 1 in 100 dilution and a 1 in 20000 dilution respectively. Blots were incubated with Goat anti-Mouse IgG H&L (IRDye® 800CW) preabsorbed (ab216772) and Goat anti-Rabbit IgG H&L (IRDye® 680RD) preabsorbed (ab216777) secondary antibodies at 1 in 20000 dilution for 1 h at room temperature before imaging.

All lanes:

Mouse monoclonal antibody at 1/100 dilution

Lane 1:

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

Lane 2:

SH3GL1 knockout HEK-293T cell lysate at 20 µg

Lane 2:

Western blot - Human SH3GL1 (EEN) knockout HEK-293T cell line (<a href='/en-us/products/cell-lines/human-sh3gl1-een-knockout-hek-293t-cell-line-ab266260'>ab266260</a>)

Lane 3:

HUVEC cell lysate at 20 µg

Lane 4:

A-431 cell lysate at 20 µg

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Sanger Sequencing - Human SH3GL1 (EEN) knockout HEK-293T cell lysate (AB259109)
  • Sanger seq

Unknown

Sanger Sequencing - Human SH3GL1 (EEN) knockout HEK-293T cell lysate (AB259109)

Allele-1 : 10 bp deletion in exon 2

Sanger Sequencing - Human SH3GL1 (EEN) knockout HEK-293T cell lysate (AB259109)
  • Sanger seq

Unknown

Sanger Sequencing - Human SH3GL1 (EEN) knockout HEK-293T cell lysate (AB259109)

Allele-2 : Insertion of the selection cassette in exon 2

Key facts

Cell type

HEK-293T

Species or organism

Human

Tissue

Kidney

Knockout validation

Sanger Sequencing,Western blot

Mutation description

Knockout achieved by using CRISPR/Cas9, 10 bp deletion in exon 2 and Insertion of the selection cassette in exon 2.

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Complementary Products

Cell Lines & Lysates

AB266260

Human SH3GL1 (EEN) knockout HEK-293T cell line

Advanced Validation

Western blot - Human SH3GL1 (EEN) knockout HEK-293T cell line (AB266260)

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Primary Antibodies

AB317836

Anti-SH3GL1/Endophilin A2 antibody [K51/1] - BSA and Azide free

BOND RX™ Validated|Recombinant

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-SH3GL1/Endophilin A2 antibody [K51/1] - BSA and Azide free (AB317836)

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Reactivity data

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

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, 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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What's included?

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Properties and storage information

Gene name
SH3GL1
Gene editing type
Knockout
Gene editing method
CRISPR technology
Knockout validation
Sanger Sequencing, Western blot
Shipped at conditions
Ambient - Can Ship with Ice
Appropriate short-term storage conditions
-20°C
Appropriate long-term storage conditions
-20°C
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Supplementary information

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

EEN also known as Endophilin A2 is a protein involved in endocytosis and membrane trafficking. EEN has a molecular weight of approximately 44 kDa and is mentioned in several tissues including the brain where it shows high expression levels. This protein aids in clathrin-mediated endocytosis by binding to lipids and proteins playing a critical role in membrane dynamics. EEN interacts with SH3 domain-containing proteins contributing to its function in cellular processes.
Biological function summary

EEN participates in the formation of protein complexes necessary for synaptic vesicle recycling important for neurotransmitter release in neurons. It binds to proline-rich motifs on its partner proteins which helps in vesicle scission during endocytosis. These interactions support vesicle fission and transport making EEN important for maintaining efficient synaptic transmission. Apart from neurons EEN also contributes to endocytic processes in other cell types indicating its wider functional role.

Pathways

EEN plays a significant role in the regulation of intracellular trafficking pathways and is a part of the endocytic pathway. It links with proteins such as dynamin and clathrin integral components of the endocytic machinery that facilitate vesicle budding and release. These interactions ensure proper recycling of synaptic vesicles and membrane components vital for cellular homeostasis. Collaborations with proteins like Bin1 show EEN's involvement in coordinating endocytosis and membrane curvature recognition.

EEN associates with conditions related to neurological function and cancer development. Aberrant expression of EEN contributes to certain cancers where disturbances in endocytosis and signaling pathways can lead to uncontrolled cell growth. The interaction with proteins such as c-Cbl an E3 ubiquitin-protein ligase implicates EEN in pathways relevant to tumor progression. Additionally altered EEN function might impact neurodegenerative diseases stemming from its contribution to synaptic vesicle trafficking reflecting the importance of its proper regulation in maintaining cell functions.
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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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Product protocols

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Product promise

We are committed to supporting your work with high-quality reagents, and we're here for you every step of the way. In the unlikely event that one of our products does not perform as expected, you're protected by our Product Promise.
For full details, please see our Terms & Conditions

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

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