Anti-B7-H3/CD276 Antibody, clone 2E22 ZooMAb® Rabbit Monoclonal
SIGMA/ZRB1826 - recombinant, expressed in HEK 293 cells
Product Type: Chemical
Immunohistochemistry (Paraffin) Enhanced Validation - Recombinant Antibody Technology Formalin Fixed Paraffin Embedded (FFPE) human tonsil (A and B) and Negative control (no primary) (C and D) tissue sections were prepared using heat-induced epitope retrieval (HIER). Immunostaining was performed using a 1:100 dilution of Cat. No. ZRB1826, Anti-B7-H3/CD276, clone 2E22 ZooMAb® Rabbit Monoclonal. Reactivity was detected using an Goat Anti-Rabbit IgG and HRP-DAB. Membranous staining was primarily observed in germinal centers, with a lesser frequency of staining present in venules & epithelium of human tonsil tissue sections.
Immunohistochemistry (Paraffin) Enhanced Validation - Recombinant Antibody Technology Formalin Fixed Paraffin Embedded (FFPE) human tonsil (A and B) and Negative control (no primary) (C and D) tissue sections were prepared using heat-induced epitope retrieval (HIER). Immunostaining was performed using a 1:100 dilution of Cat. No. ZRB1826, Anti-B7-H3/CD276, clone 2E22 ZooMAb® Rabbit Monoclonal. Reactivity was detected using an Goat Anti-Rabbit IgG and HRP-DAB. Membranous staining was primarily observed in germinal centers, with a lesser frequency of staining present in venules & epithelium of human tonsil tissue sections.
Immunohistochemistry (Paraffin) Enhanced Validation - Recombinant Antibody Technology Formalin Fixed Paraffin Embedded (FFPE) human tonsil (A and B) and Negative control (no primary) (C and D) tissue sections were prepared using heat-induced epitope retrieval (HIER). Immunostaining was performed using a 1:100 dilution of Cat. No. ZRB1826, Anti-B7-H3/CD276, clone 2E22 ZooMAb® Rabbit Monoclonal. Reactivity was detected using an Goat Anti-Rabbit IgG and HRP-DAB. Membranous staining was primarily observed in germinal centers, with a lesser frequency of staining present in venules & epithelium of human tonsil tissue sections.
Immunohistochemistry (Paraffin) Enhanced Validation - Recombinant Antibody Technology Formalin Fixed Paraffin Embedded (FFPE) human tonsil (A and B) and Negative control (no primary) (C and D) tissue sections were prepared using heat-induced epitope retrieval (HIER). Immunostaining was performed using a 1:100 dilution of Cat. No. ZRB1826, Anti-B7-H3/CD276, clone 2E22 ZooMAb® Rabbit Monoclonal. Reactivity was detected using an Goat Anti-Rabbit IgG and HRP-DAB. Membranous staining was primarily observed in germinal centers, with a lesser frequency of staining present in venules & epithelium of human tonsil tissue sections.
Immunohistochemistry (Paraffin) Enhanced Validation - Recombinant Antibody Technology Formalin Fixed Paraffin Embedded (FFPE) human tonsil (A and B) and Negative control (no primary) (C and D) tissue sections were prepared using heat-induced epitope retrieval (HIER). Immunostaining was performed using a 1:100 dilution of Cat. No. ZRB1826, Anti-B7-H3/CD276, clone 2E22 ZooMAb® Rabbit Monoclonal. Reactivity was detected using an Goat Anti-Rabbit IgG and HRP-DAB. Membranous staining was primarily observed in germinal centers, with a lesser frequency of staining present in venules & epithelium of human tonsil tissue sections.
Immunocytochemistry Enhanced Validation - Recombinant Antibody Technology Immunofluorescent analysis of U251 cell line was performed using a 1:100 dilution of Cat. No. ZRB1826, Anti-B7-H3/CD276, clone 2E22 ZooMAb® Rabbit Monoclonal and visualized with a Goat Anti-Rabbit secondary antibody conjugated to Alexa FluorZooMAb® 488 (Green). Actin filaments have been labeled with phalloidin (Red). Nucleus is stained with DAPI (Blue). This antibody positively stains the membrane.
Immunocytochemistry Enhanced Validation - Recombinant Antibody Technology Immunofluorescent analysis of U251 cell line was performed using a 1:100 dilution of Cat. No. ZRB1826, Anti-B7-H3/CD276, clone 2E22 ZooMAb® Rabbit Monoclonal and visualized with a Goat Anti-Rabbit secondary antibody conjugated to Alexa FluorZooMAb® 488 (Green). Actin filaments have been labeled with phalloidin (Red). Nucleus is stained with DAPI (Blue). This antibody positively stains the membrane.
Immunocytochemistry Enhanced Validation - Recombinant Antibody Technology Immunofluorescent analysis of U251 cell line was performed using a 1:100 dilution of Cat. No. ZRB1826, Anti-B7-H3/CD276, clone 2E22 ZooMAb® Rabbit Monoclonal and visualized with a Goat Anti-Rabbit secondary antibody conjugated to Alexa FluorZooMAb® 488 (Green). Actin filaments have been labeled with phalloidin (Red). Nucleus is stained with DAPI (Blue). This antibody positively stains the membrane.
Western Blotting Enhanced Validation - Recombinant Antibody Technology Lysates from U251 (Lane 1) and PC3 (Lane 2) cells were probed with Cat. No. ZRB1826, Anti-B7-H3/CD276, clone 2E22 ZooMAb® Rabbit Monoclonal (1:1,000 dilution). Proteins were visualized using a Donkey Anti-Rabbit IgG secondary antibody conjugated to HRP and a chemiluminescence detection system. Arrow indicates B7-H3/CD276 (~90 kDa).
Affinity Binding Assay Enhanced Validation - Recombinant Antibody Technology 100 μg/mL of non-specific control peptide (Run 1) or B7-H3/CD276 peptide (Run 2) were analyzed by affinity binding assay using Cat. No. ZRB1826, Anti-B7-H3/CD276, clone 2E22 ZooMAb® Rabbit Monoclonal. The binding of this antibody to B7-H3/CD276 peptide displayed a KD of 7.2 x 10-8 whereas binding to non-specific control peptide displayed a KD of 3.1 x 10-3.
12 Principles of Green Chemistry: Principle 1—Waste Prevention. This product has a smaller environmental footprint through reduction of waste in the manufacturing process.
12 Principles of Green Chemistry: Principle 4—Designing Safer Chemicals
Chemical products should be designed to affect their desired function while minimizing their toxicity.
Chemical products should be designed to affect their desired function while minimizing their toxicity.
12 Principles of Green Chemistry: Principle 6—Design for Energy Efficiency. This product was re-engineered to utilize less energy in the manufacturing process.
| antibody form | purified antibody |
| antibody product type | primary antibodies |
| biological source | rabbit |
| clone | 2E22, recombinant monoclonal |
| conjugate | unconjugated |
| description | recombinant, expressed in HEK 293 cells |
| enhanced validation | recombinant expression Learn more about Antibody Enhanced Validation  |
| epitope sequence | N-terminal half |
| form | lyophilized |
| greener alternative category | Aligned |
| greener alternative product characteristics | Waste Prevention Designing Safer Chemicals Design for Energy Efficiency Learn more about the Principles of Green Chemistry . |
| isotype | IgG |
| mol wt | calculated mol wt 57.24 kDa |
| observed mol wt ~90 kDa | |
| packaging | antibody small pack of 25 μL |
| product line | ZooMAb® learn more |
| Protein ID accession no. | NP_001019907  |
| purified by | using Protein A |
| Quality Level | 200 |
| recombinant | expressed in HEK 293 cells |
| shipped in | ambient |
| species reactivity | human |
| species reactivity (predicted by homology) | porcine, monkey |
| storage temp. | 2-8°C |
| technique(s) | affinity binding assay: suitable |
| immunocytochemistry: suitable | |
| immunohistochemistry (formalin-fixed, paraffin-embedded sections): suitable | |
| western blot: suitable | |
| UniProt accession no. | Q5ZPR3 |
| Application: | Quality Control Testing Evaluated by Western Blotting in U251 cell lysate. Western Blotting Analysis: A 1:1,000 dilution of this antibody detected B7-H3/CD276 in U251 cell lysate. Tested applications Western Blotting Analysis: A 1:1,000 dilution from a representative lot detected B7-H3/CD276 in PC3 cell lysate. Immunocytochemistry Analysis: A 1:100 dilution from a representative lot detected B7-H3/CD276 in U251 cells. Affinity Binding Assay: A representative lot of this antibody bound CD276 (B7-H3) with a KD of 7.2 x 10-8 in an affinity binding assay. Immunohistochemistry (Paraffin) Analysis: A 1:100 dilution from a representative lot detected B7-H3/CD276 in human tonsil tissue sections. Note: Actual optimal working dilutions must be determined by end user as specimens, and experimental conditions may vary with the end user |
| Disclaimer: | Unless otherwise stated in our catalog or other company documentation accompanying the product(s), our products are intended for research use only and are not to be used for any other purpose, which includes but is not limited to, unauthorized commercial uses, in vitro diagnostic uses, ex vivo or in vivo therapeutic uses or any type of consumption or application to humans or animals. |
| General description: | We are committed to bringing you greener alternative products, which adhere to one or more of The 12 Principles of Green Chemistry.This antibody is Preservative-free, produced without the harm or sacrifice of animals and exceptionally stable to allow for ambient shipping and storage if needed and thus aligns with "Waste Prevention", "Designing Safer Chemicals" and "Design for Energy Efficiency". Click here for more information. |
| General description: | ZooMAb® antibodies represent an entirely new generation of recombinant monoclonal antibodies.Each ZooMAb® antibody is manufactured using our proprietary recombinant expression system, purified to homogeneity, and precisely dispensed to produce robust and highly reproducible lot-to-lot consistency. Only top-performing clones are released for use by researchers. Each antibody is validated for high specificity and affinity across multiple applications, including its most commonly used application. ZooMAb® antibodies are reliably available and ready to ship when you need them. |
| Immunogen: | KLH-conjugated linear peptide corresponding to 21 amino acids from the N-terminal half of human B7-H3/CD276. |
| Legal Information: | ZooMAb is a registered trademark of Merck KGaA, Darmstadt, Germany |
| Physical form: | Purified recombinant rabbit monoclonal antibody IgG, lyophilized in PBS, 5% Trehalose, normal appearance a coarse or translucent resin. The PBS/trehalose components in the ZooMAb formulation can have the appearance of a semi-solid (bead like gel) after lyophilization. This is a normal phenomenon. Please follow the recommended reconstitution procedure in the data sheet to dissolve the semi-solid, bead-like, gel-appearing material. The resulting antibody solution is completely stable and functional as proven by full functional testing. Contains no biocide or preservatives, such as azide, or any animal by-products. Larger pack sizes provided as multiples of 25 μL. |
| Reconstitution: | 300 μg/mL after reconstitution at 25 μL per vial. Please refer to guidance on suggested starting dilutions and/or titers per application and sample type. |
| Specificity: | Clone 2E22 is a ZooMAb® Rabbit recombinant monoclonal antibody that specifically detects B7-H3/CD276. It targets an epitope within 21 amino acids from the extracellular domain within the N-terminal half. |
| Storage and Stability: | Recommend storage of lyophilized product at 2-8°C; Before reconstitution, micro-centrifuge vials briefly to spin down material to bottom of the vial; Reconstitute each vial by adding 25 μL of filtered lab grade water or PBS; Reconstituted antibodies can be stored at 2-8°C, or -20°C for long term storage. Avoid repeated freeze-thaws. |
| Target description: | CD276 antigen (UniProt: UniProt: Q5ZPR3; also known as 4Ig-B7-H3, B7 homolog 3, B7-H3, Costimulatory molecule, CD276) is encoded by the CD276 (also known as B7H3, PSEC0249, UNQ309/PRO352) gene (Gene ID: 80381) in human. CD276 is a single-pass type I membrane protein that is member of B7 immunoregulatory family. It is synthesized with a signal peptide (aa 1-28), which is subsequently cleaved off to produce the mature form that contains an extracellular domain (aa 29-466), a short transmembrane domain (aa 467-487), and a cytoplasmic domain (aa 488-534). CD276 participates in the regulation of T-cell-mediated immune response. It is a ubiquitous protein but is undetectable in peripheral blood lymphocytes or granulocytes and is only weakly expressed in resting monocytes. It is also expressed in dendritic cells derived from monocytes and in epithelial cells of sinonasal tissue, extravillous trophoblast cells, and Hofbauer cells of the first trimester placenta and term placenta. It plays a key role in providing the placenta and fetus with a suitable immunological environment throughout pregnancy. Higher expression of CD276 has been shown in a variety of cancer types, including breast, pancreatic, and ovarian cancer and its expression has been correlated with poor survival in ovarian cancer subjects. It plays a protective role in tumor cells by inhibiting natural-killer mediated cell lysis. Its levels are also reported to be up-regulated in cells that mediate rejection of human transplants. This ZooMAbZooMAb® recombinant monoclonal antibody, generated by our propriety technology, offers significantly enhanced specificity, affinity, reproducibility, and stability over conventional monoclonals. (Ref.: Chen, Y-W., et al. (2008). Curr. Can. Drug Targets. 8(5); 404-413; Wang, L., et al. (2005). Eur. J. Immunol. 35(2); 428-438; Chapoval, AI., et al. (2001). Nat. Immunol. 2(3); 269-274). |
| Storage Temp. | 2-8°C |
| UNSPSC | 12352203 |