Product Info Summary
SKU: | A02220-3 |
---|---|
Size: | 100 µg/vial |
Reactive Species: | Human |
Host: | Rabbit |
Application: | ELISA, Flow Cytometry, IHC, WB |
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Product info
Product Name
Anti-CD276 Antibody Picoband™
View all B7-H3/CD276 Antibodies
SKU/Catalog Number
A02220-3
Size
100 µg/vial
Form
Lyophilized
Description
Boster Bio Anti-CD276 Antibody Picoband™ catalog # A02220-3. Tested in ELISA, Flow Cytometry, IHC, WB applications. This antibody reacts with Human.
Storage & Handling
At -20°C for one year from date of receipt. After reconstitution, at 4°C for one month. It can also be aliquotted and stored frozen at -20°C for six months. Avoid repeated freezing and thawing.
Cite This Product
Anti-CD276 Antibody Picoband™ (Boster Biological Technology, Pleasanton CA, USA, Catalog # A02220-3)
Host
Rabbit
Contents
Each vial contains 4 mg Trehalose, 0.9 mg NaCl, 0.2 mg Na2HPO4.
Clonality
Polyclonal
Isotype
Rabbit IgG
Immunogen
E.coli-derived human CD276 recombinant protein (Position: L29-Q458).
*Blocking peptide can be purchased. Costs vary based on immunogen length. Contact us for pricing.
Cross-reactivity
No cross-reactivity with other proteins.
Reactive Species
A02220-3 is reactive to CD276 in Human
Applications
A02220-3 is guaranteed for ELISA, Flow Cytometry, IHC, WB Boster Guarantee
Observed Molecular Weight
90-100 kDa
Calculated molecular weight
57.235kDa
Background of B7-H3/CD276
Cluster of Differentiation 276 (CD276) or B7 Homolog 3 (B7-H3) is a human protein encoded by the CD276 gene. The protein encoded by this gene belongs to the immunoglobulin superfamily, and thought to participate in the regulation of T-cell-mediated immune response. Studies show that while the transcript of this gene is ubiquitously expressed in normal tissues and solid tumors, the protein is preferentially expressed only in tumor tissues. Additionally, it was observed that the 3' UTR of this transcript contains a target site for miR29 microRNA, and there is an inverse correlation between the expression of this protein and miR29 levels, suggesting regulation of expression of this gene product by miR29. Alternatively spliced transcript variants encoding different isoforms have been found for this gene.
Antibody Validation
Boster validates all antibodies on WB, IHC, ICC, Immunofluorescence, and ELISA with known positive control and negative samples to ensure specificity and high affinity, including thorough antibody incubations.
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Assay dilution & Images
Reconsitution
Adding 0.2 ml of distilled water will yield a concentration of 500 µg/ml.
Assay Dilutions Recommendation
The recommendations below provide a starting point for assay optimization. The actual working concentration varies and should be decided by the user.
Western blot, 0.25-0.5 µg/ml, Human
Immunohistochemistry(Paraffin-embedded Section), 2-5 µg/ml, Human
Flow Cytometry, 1-3 µg/1x106 cells, Human
Direct ELISA, 0.1-0.5 µg/ml, Human
Validation Images & Assay Conditions
Click image to see more details
Figure 1. Western blot analysis of CD276 using anti-CD276 antibody (A02220-3).
Electrophoresis was performed on a 5-20% SDS-PAGE gel at 70V (Stacking gel) / 90V (Resolving gel) for 2-3 hours. The sample well of each lane was loaded with 30 ug of sample under reducing conditions.
Lane 1: human U2OS whole cell lysates,
Lane 2: human PC-3 whole cell lysates,
Lane 3: human Hela whole cell lysates,
Lane 4: human MCF-7 whole cell lysates.
After electrophoresis, proteins were transferred to a nitrocellulose membrane at 150 mA for 50-90 minutes. Blocked the membrane with 5% non-fat milk/TBS for 1.5 hour at RT. The membrane was incubated with rabbit anti-CD276 antigen affinity purified polyclonal antibody (Catalog # A02220-3) at 0.5 μg/mL overnight at 4°C, then washed with TBS-0.1%Tween 3 times with 5 minutes each and probed with a goat anti-rabbit IgG-HRP secondary antibody at a dilution of 1:5000 for 1.5 hour at RT. The signal is developed using an Enhanced Chemiluminescent detection (ECL) kit (Catalog # EK1002) with Tanon 5200 system. A specific band was detected for CD276 at approximately 90-100 kDa. The expected band size for CD276 is at 100 kDa.
Click image to see more details
Figure 2. IHC analysis of CD276 using anti-CD276 antibody (A02220-3).
CD276 was detected in a paraffin-embedded section of human rectal cancer tissue. Heat mediated antigen retrieval was performed in EDTA buffer (pH 8.0, epitope retrieval solution). The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 2 μg/ml rabbit anti-CD276 Antibody (A02220-3) overnight at 4°C. Peroxidase Conjugated Goat Anti-rabbit IgG was used as secondary antibody and incubated for 30 minutes at 37°C. The tissue section was developed using HRP Conjugated Rabbit IgG Super Vision Assay Kit (Catalog # SV0002) with DAB as the chromogen.
Click image to see more details
Figure 3. IHC analysis of CD276 using anti-CD276 antibody (A02220-3).
CD276 was detected in a paraffin-embedded section of human liver cancer tissue. Heat mediated antigen retrieval was performed in EDTA buffer (pH 8.0, epitope retrieval solution). The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 2 μg/ml rabbit anti-CD276 Antibody (A02220-3) overnight at 4°C. Peroxidase Conjugated Goat Anti-rabbit IgG was used as secondary antibody and incubated for 30 minutes at 37°C. The tissue section was developed using HRP Conjugated Rabbit IgG Super Vision Assay Kit (Catalog # SV0002) with DAB as the chromogen.
Click image to see more details
Figure 4. IHC analysis of CD276 using anti-CD276 antibody (A02220-3).
CD276 was detected in a paraffin-embedded section of human tonsil tissue. Heat mediated antigen retrieval was performed in EDTA buffer (pH 8.0, epitope retrieval solution). The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 2 μg/ml rabbit anti-CD276 Antibody (A02220-3) overnight at 4°C. Peroxidase Conjugated Goat Anti-rabbit IgG was used as secondary antibody and incubated for 30 minutes at 37°C. The tissue section was developed using HRP Conjugated Rabbit IgG Super Vision Assay Kit (Catalog # SV0002) with DAB as the chromogen.
Click image to see more details
Figure 5. Flow Cytometry analysis of PC-3 cells using anti-CD276 antibody (A02220-3).
Overlay histogram showing PC-3 cells stained with A02220-3 (Blue line). The cells were blocked with 10% normal goat serum. And then incubated with rabbit anti-CD276 Antibody (A02220-3, 1 μg/1x106 cells) for 30 min at 20°C. DyLight®488 conjugated goat anti-rabbit IgG (BA1127, 5-10 μg/1x106 cells) was used as secondary antibody for 30 minutes at 20°C. Isotype control antibody (Green line) was rabbit IgG (1 μg/1x106) used under the same conditions. Unlabelled sample (Red line) was also used as a control.
Protein Target Info & Infographic
Gene/Protein Information For CD276 (Source: Uniprot.org, NCBI)
Gene Name
CD276
Full Name
CD276 antigen
Weight
57.235kDa
Superfamily
immunoglobulin superfamily
Alternative Names
B7H3; B7-H3; B7H34Ig-B7-H3; B7-H3B7 homolog 3; CD276 antigen; CD276 molecule; CD276; Costimulatory molecule CD276 4Ig-B7-H3, B7-H3, B7H3, B7RP-2 CD276 molecule CD276 antigen|B7 homolog 3|costimulatory molecule
*If product is indicated to react with multiple species, protein info is based on the gene entry specified above in "Species".For more info on CD276, check out the CD276 Infographic
We have 30,000+ of these available, one for each gene! Check them out.
In this infographic, you will see the following information for CD276: database IDs, superfamily, protein function, synonyms, molecular weight, chromosomal locations, tissues of expression, subcellular locations, post-translational modifications, and related diseases, research areas & pathways. If you want to see more information included, or would like to contribute to it and be acknowledged, please contact [email protected].
Specific Publications For Anti-CD276 Antibody Picoband™ (A02220-3)
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