Human CD276 knockout HeLa cell lysate
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- WB
Lab
Western blot - Human CD276 knockout HeLa cell lysate (AB257098)
Lane 1 : Wild-type HeLa cell lysate (20µg)
Lane 2 : CD276 knockout HeLa cell lysate (20µg)
Lane 3 : LNCaP cell lysate (20µg)
Lane 4 : Raji cell lysate (20µg)
Lanes 1- 4 : Merged signal (red and green). Green - ab227670 observed at 90-110 kDa. Red - loading control ab8245 observed at 37 kDa.
ab227670 Rabbit monoclonal [SP206] to CD276 was shown to specifically react with CD276 antigen in wild-type HeLa cells in western blot. Loss of signal was observed when knockout cell line ab261756 (knockout cell lysate ab257098) was used. Wild-type and CD276 antigen 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. ab227670 and Anti-GAPDH antibody [6C5] - Loading Control (ab8245) were incubated overnight at 4°C at 1 in 400 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.
Lanes 1 - 4:
Western blot - Anti-CD276 antibody [SP206], prediluted (<a href='/en-us/products/primary-antibodies/cd276-antibody-sp206-prediluted-ab228178'>ab228178</a>) at 1/400 dilution
Lanes 1 - 4:
Western blot - Anti-CD276 antibody [SP206] (<a href='/en-us/products/primary-antibodies/cd276-antibody-sp206-ab227670'>ab227670</a>) at 1/400 dilution
Lane 1:
Wild-type HeLa cell lysate at 20 µg
Lane 2:
CD276 knockout HeLa cell lysate at 20 µg
Lane 2:
Western blot - Human CD276 knockout HeLa cell line (<a href='/en-us/products/cell-lines/human-cd276-knockout-hela-cell-line-ab261756'>ab261756</a>)
Lane 3:
LNCaP cell lysate at 20 µg
Lane 4:
Raji cell lysate at 20 µg
Predicted band size: 57 kDa
Observed band size: 90-110 kDa
false
- Sanger seq
Unknown
Sanger Sequencing - Human CD276 knockout HeLa cell lysate (AB257098)
Homozygous : 11 bp deletion in exon 2
Complementary Products
![Western blot - Anti-CD276 antibody [EPNCIR122] (AB134161)](https://content.abcam.com/products/images/cd276-antibody-epncir122-ab134161--western-blot-img155687.jpg)
Reactivity data
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.
What's included?
Properties and storage information
Gene name
Gene editing type
Gene editing method
Knockout validation
Zygosity
Shipped at conditions
Appropriate short-term storage conditions
Appropriate long-term storage conditions
Supplementary information
This supplementary information is collated from multiple sources and compiled automatically.
Biological function summary
CD276 modulates immune responses by acting as a co-inhibitory ligand impacting T-cell proliferation and cytokine synthesis. Not part of a known complex it plays a role in the adaptive immune system by providing negative feedback regulation that can inhibit the immune response. This capability has implications in preventing autoimmunity but may also contribute to tumor immune evasion by reducing effective anti-tumor responses. The B7-H3/CD276 interaction with receptors such as IL receptor family members suggests its diverse immunomodulatory functions.
Pathways
CD276 participates in the T-cell receptor (TCR) signaling pathways influencing immune checkpoint pathways. It interacts with other immune checkpoint proteins like PD-L1 and CTLA-4 but its exact receptor partner remains unidentified. The presence of CD276 in these pathways highlights its role in controlling T-cell activation and tolerance. By modulating TCR signals and impacting downstream effects CD276 serves as a regulatory point that can balance immune activation and inhibition critical in immune-mediated conditions.
Quality control
STR analysis
CSF1PO, D13S317, D7S820, D5S818, TH01, D16S539, TPOX
Cell culture
Biosafety level
EU: 2 US: 2
Adherent/suspension
Adherent
Gender
Female
Product promise
Please note: All products are 'FOR RESEARCH USE ONLY. NOT FOR USE IN DIAGNOSTIC OR THERAPEUTIC PROCEDURES'.
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