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AB257220

Human CD47 knockout HEK-293T cell lysate

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CD47 KO cell lysate available now. KO validated by Western blot. Free of charge wild type control included. Knockout achieved by using CRISPR/Cas9, 11 bp deletion in exon 2 and 5 bp deletion in exon 2.
3 Images
Western blot - Human CD47 knockout HEK-293T cell lysate (AB257220)
  • WB

Lab

Western blot - Human CD47 knockout HEK-293T cell lysate (AB257220)

Lane 1 : Wild-type HEK293T cell lysate (20 ug)

Lane 2 : CD47 knockout HEK293T cell lysate (20 ug)

Lane 3 : Jurkat cell lysate (20 ug)

Lane 4 : HepG2 cell lysate (20 ug)

Lanes 1-4 : Merged signal (red and green). Green - ab218810 observed at 47-52 kDa. Red - loading control ab8245 observed at 36 kDa.

ab218810 Anti-CD47 antibody [EPR21794] was shown to specifically react with CD47 in wild-type HEK293T cells. Loss of signal was observed when knockout cell line ab266324 (knockout cell lysate ab257220) was used. Wild-type and CD47 knockout samples were subjected to SDS-PAGE. ab218810 and Anti-GAPDH antibody [6C5] - Loading Control (ab8245) were incubated overnight at 4°C at 1 in 500 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-CD47 antibody [EPR21794] (<a href='/en-us/products/primary-antibodies/cd47-antibody-epr21794-ab218810'>ab218810</a>) at 1/500 dilution

Lane 1:

Wild-type HEK293T cell lysate at 20 µg

Lane 2:

CD47 knockout HEK293T cell lysate at 20 µg

Lane 2:

Western blot - Human CD47 knockout HEK-293T cell line (<a href='/en-us/products/cell-lines/human-cd47-knockout-hek-293t-cell-line-ab266324'>ab266324</a>)

Lane 3:

Jurkat cell lysate at 20 µg

Lane 4:

HepG2 cell lysate at 20 µg

Secondary

All lanes:

Western blot - Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed (<a href='/en-us/products/secondary-antibodies/goat-rabbit-igg-h-l-irdye-800cw-preadsorbed-ab216773'>ab216773</a>) at 1/10000 dilution

Predicted band size: 35 kDa

Observed band size: 47-52 kDa

false

Sanger Sequencing - Human CD47 knockout HEK-293T cell lysate (AB257220)
  • Sanger seq

Unknown

Sanger Sequencing - Human CD47 knockout HEK-293T cell lysate (AB257220)

Allele-2 : 5 bp deletion in exon 2

Sanger Sequencing - Human CD47 knockout HEK-293T cell lysate (AB257220)
  • Sanger seq

Unknown

Sanger Sequencing - Human CD47 knockout HEK-293T cell lysate (AB257220)

Allele-1 : 11 bp deletion 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, 11 bp deletion in exon 2 and 5 bp deletion in exon 2.

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Reactivity data

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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 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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What's included?

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Properties and storage information

Gene name
CD47
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.

CD47 also referred to as integrin-associated protein carries a molecular weight of approximately 50 kDa. This protein is broadly expressed across various cell types notably on erythrocytes leukocytes and endothelial cells. Its transmembrane glycoprotein structure allows it to perform various cellular functions. CD47 interacts with specific ligands prominently SIRPα which are present on immune cells such as macrophages and dendritic cells. The engagement of CD47 with these ligands plays a major role in cellular signaling processes.
Biological function summary

CD47 functions as a "don't eat me" signal inhibiting phagocytosis by binding to SIRPα on macrophages. This protein is also important for cell adhesion and migration and can modulate interactions with integrins. While not a part of a large complex CD47 associates with various cytoskeletal and membrane proteins to maintain cellular architecture and transmission of mechanical forces. Additionally its interaction with thrombospondins influences angiogenesis and nitric oxide signaling.

Pathways

CD47 is extensively involved in the regulation of the immune response and angiogenesis. The interaction between CD47 and SIRPα plays a part in the regulation of macrophage activity impacting the immune signaling pathway. CD47 also interacts with integrins allowing it to contribute to the angiogenesis pathway which is critical in the formation of new blood vessels. These interactions help coordinate complex cellular responses and maintain tissue homeostasis.

CD47 levels have associations with cancer and chronic inflammatory diseases. In cancer CD47 overexpression can allow tumor cells to evade the immune response by downregulating phagocytosis through SIRPα interaction. Therapeutically targeting CD47 using agents like anti-CD47 antibodies shows potential for enhancing anti-tumor immunity. Additionally CD47 is involved in atherosclerosis where its interaction with thrombospondin-1 contributes to disease pathogenesis by affecting nitric oxide signaling and vascular remodeling.
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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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