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AB257097

Human CD276 knockout HEK-293T cell lysate

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CD276 KO cell lysate available now. KO validated by Western blot. Free of charge wild type control included. Knockout achieved by using CRISPR/Cas9, Homozygous: Insertion of the selection cassette in exon 2.
3 Images
Western blot - Human CD276 knockout HEK-293T cell lysate (AB257097)
  • WB

Lab

Western blot - Human CD276 knockout HEK-293T cell lysate (AB257097)

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

Lane 2 : CD276 knockout HEK293T cell lysate (20 μg)

Lane 3 : LNCaP cell lysate (20 μg)

Lane 4 : Raji cell lysate (20 μg)

Lanes 1-4 : Merged signal (red and green). Green - ab227670 observed at 90-110 kDa. Red - loading control ab8245 observed at 36 kDa.

ab227670 Anti-CD276 antibody [SP206] was shown to specifically react with CD276 in wild-type HEK293T cells. Loss of signal was observed when knockout cell line ab266658 (knockout cell lysate ab257097) was used. Wild-type and CD276 knockout samples were subjected to SDS-PAGE. ab227670 and Anti-GAPDH antibody [6C5] - Loading Control (ab8245) were incubated overnight at 4°C at 1 in 1000 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] (<a href='/en-us/products/primary-antibodies/cd276-antibody-sp206-ab227670'>ab227670</a>) at 1/1000 dilution

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 1/1000 dilution

Lane 1:

Wild-type HEK293T cell lysate at 20 µg

Lane 2:

CD276 knockout HEK293T cell lysate at 20 µg

Lane 2:

Western blot - Human CD276 knockout HEK-293T cell line (<a href='/en-us/products/cell-lines/human-cd276-knockout-hek-293t-cell-line-ab266658'>ab266658</a>)

Lane 3:

LNCaP cell lysate at 20 µg

Lane 4:

Raji 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: 57 kDa

Observed band size: 90-110 kDa

false

Western blot - Human CD276 knockout HEK-293T cell lysate (AB257097)
  • WB

Lab

Western blot - Human CD276 knockout HEK-293T cell lysate (AB257097)

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

Lane 2 : CD276 knockout HEK293T cell lysate (20 μg)

Lane 3 : LNCaP cell lysate (20 μg)

Lane 4 : Raji cell lysate (20 μg)

Lanes 1-4 : Merged signal (red and green). Green - ab219648 observed at 90-110 kDa. Red - loading control ab8245 observed at 36 kDa.

ab219648 Anti-CD276 antibody [EPR20115] was shown to specifically react with CD276 in wild-type HEK293T cells. Loss of signal was observed when knockout cell line ab266658 (knockout cell lysate ab257097) was used. Wild-type and CD276 knockout samples were subjected to SDS-PAGE. ab219648 and Anti-GAPDH antibody [6C5] - Loading Control (ab8245) were incubated overnight at 4°C at 1 in 1000 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-CD276 antibody [EPR20115] (<a href='/en-us/products/primary-antibodies/cd276-antibody-epr20115-ab219648'>ab219648</a>) at 1/1000 dilution

Lane 1:

Wild-type HEK293T cell lysate at 20 µg

Lane 2:

CD276 knockout HEK293T cell lysate at 20 µg

Lane 2:

Western blot - Human CD276 knockout HEK-293T cell line (<a href='/en-us/products/cell-lines/human-cd276-knockout-hek-293t-cell-line-ab266658'>ab266658</a>)

Lane 3:

LNCaP cell lysate at 20 µg

Lane 4:

Raji 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: 57 kDa

Observed band size: 90-110 kDa

false

Sanger Sequencing - Human CD276 knockout HEK-293T cell lysate (AB257097)
  • Sanger seq

Unknown

Sanger Sequencing - Human CD276 knockout HEK-293T cell lysate (AB257097)

Homozygous : 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, Homozygous: Insertion of the selection cassette in exon 2.

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Complementary Products

Cell Lines & Lysates

AB266658

Human CD276 knockout HEK-293T cell line

Advanced Validation

Western blot - Human CD276 knockout HEK-293T cell line (AB266658)

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Primary Antibodies

AB134161

Anti-CD276 antibody [EPNCIR122]

RabMAb|Recombinant|KO Validated|20ul selling size

Western blot - Anti-CD276 antibody [EPNCIR122] (AB134161)

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Proteins & Peptides

AB134865

Recombinant Human CD276 protein

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  • 0
Proteins & Peptides

AB288799

Recombinant Human VCAM1 protein (Active)

Premium Bioactive

Biological Activity - Recombinant Human VCAM1 protein (Active) (AB288799)

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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, 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
CD276
Gene editing type
Knockout
Gene editing method
CRISPR technology
Knockout validation
Sanger Sequencing, Western blot
Zygosity
Homozygous
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.

CD276 also known as B7-H3 is an immunoregulatory protein with a mass of approximately 57 kDa although post-translational modifications can increase its observed size. It is a member of the B7 family which is part of the immunoglobulin superfamily. CD276 is widely expressed in various tissues including placenta liver heart and also in many tumor cells. It is observed in lesser amounts in normal tissues making it an interesting target for cancer research. Various assays like HEK293 and HEK293T cell confluency studies often use CD276 to explore its expression under different conditions and seeding densities which are represented in confluency charts.
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.

CD276 has strong associations with cancer particularly in solid tumors such as prostate and breast cancer. It is often overexpressed in malignant tissues contributing to immune evasion by tumors. Researchers propose it as a potential target for cancer immunotherapy due to this attribute. CD276's link to disorders like asthma has also been suggested where its interaction with proteins involved in inflammatory pathways such as IL response elements can exacerbate the condition. This implicates it as a dual-role protein in both disease progression and potential therapeutic targeting.
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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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