CD47 Antibody (B6H12.2) - Azide and BSA Free

Immunocytochemistry/Immunofluorescence: CD47 Antibody (B6H12.2) - Azide Free [NBP2-31106] - A431 cells were fixed in 4% paraformaldehyde for 10 minutes and permeabilized in 0.05% Triton X-100 in PBS for 5 minutes. The cells were incubated with CD47 Antibody [B6H12.2] (NBP2-31106) at 1 ug/ml overnight at 4C and detected with an anti-mouse Dylight 488 (Green) at a 1:1000 dilution for 60 minutes. Nuclei were counterstained with DAPI (Blue). Cells were imaged using a 40X objective.

Western Blot: CD47 Antibody (B6H12.2) - Azide Free [NBP2-31106] - Detection of CD47 expression on two hematological cancer cell lines, Jurkat and Raji. WB image submitted by a verified customer review.

Immunohistochemistry-Paraffin: CD47 Antibody (B6H12.2) - Azide Free [NBP2-31106] - Human breast cancer tissue stained with CD47 Antibody (B6H12.2). IHC-P image submitted by a verified customer review.

Flow Cytometry: CD47 Antibody (B6H12.2) - Azide Free [NBP2-31106] - A surface stain was performed on A431 cells with CD47 Antibody [B6H12.2] NBP2-31106PCP (blue) and a matched isotype control (orange). Cells were incubated in an antibody dilution of 5 ug/mL for 20 minutes at room temperature. Both antibodies were conjugated to PerCP.

Western Blot: CD47 Antibody (B6H12.2) [Azide Free] [NBP2-31106] - Human brain (lane 1) and testis (lane 2) protein was separated on a 12% gel by SDS-PAGE. Protein was transferred to PVDF membrane, blocked and then probed with 2 ug/ml of anti-CD47. CD47 protein was detected using an anti-mouse HRP secondary antibody.

Immunocytochemistry: CD47 Antibody (B6H12.2) - Azide Free [NBP2-31106] - Human trabecular meshwork (primary ocular cells). Image from verified customer review.

Immunocytochemistry/Immunofluorescence: CD47 Antibody (B6H12.2) - Azide Free [NBP2-31106] - A431 cells were fixed in 4% paraformaldehyde for 10 minutes and permeabilized in 0.05% Triton X-100 in PBS for 5 minutes. The cells were incubated with CD47 Antibody [B6H12.2] conjugated to Biotin (NBP2-31106B) at 5 ug/ml for 1 hour at room temperature then detected with Streptavidin conjugated to DyLight 550. Nuclei were counterstained with DAPI (Blue). Cells were imaged using a 40X objective.

Immunohistochemistry-Paraffin: CD47 Antibody (B6H12.2) - Azide Free [NBP2-31106] - Tissue section of human normal breast using mouse monoclonal CD47 antibody (clone B6H12.2) at 0.5ug/ml concentration. The ductal/acinar epithelial cells in the breast section developed specific membrane-cytoplasmic staining.

Immunohistochemistry-Paraffin: CD47 Antibody (B6H12.2) - Azide Free [NBP2-31106] - Tissue section of human normal breast using mouse monoclonal CD47 antibody (clone B6H12.2) at 0.5ug/ml concentration. The ductal/acinar epithelial cells in the breast section developed specific membrane-cytoplasmic staining.

Flow (Cell Surface): CD47 Antibody (B6H12.2) - Azide Free [NBP2-31106] - A surface stain was performed on human peripheral blood lymphocytes with CD47 (B6H12.2) antibody NBP2-31106 (blue) and a matched isotype control NBP2-27287 (orange). Cells were incubated in an antibody dilution of 1 ug/mL for 20 minutes at room temperature, followed by mouse F(ab)2 IgG (H+L) APC-conjugated secondary antibody [F0101B, R&D Systems].

• Reactivity: Hu, Mu
• Applications: WB, Flow, ICC/IF, IHC, IP, CyTOF-ready
• Clone: B6H12.2
• Clonality: Monoclonal
• Host: Mouse
• Conjugate: Unconjugated
• Format: Azide and BSA Free
• Concentration: 1.0 mg/ml

CD47 Antibody (B6H12.2) - Azide and BSA Free Summary

• Immunogen: This antibody was raised against CD47.
• Isotype: IgG1
• Clonality: Monoclonal
• Host: Mouse
• Gene: CD47
• Purity: Protein G purified

Applications/Dilutions

• Dilutions:
  • CyTOF-ready
  • Flow (Cell Surface)
  • Flow Cytometry 1 ug/mL
  • Immunocytochemistry/ Immunofluorescence
  • Immunohistochemistry
  • Immunohistochemistry-Paraffin 0.5 ug/mL
  • Immunoprecipitation
  • Western Blot 1:1000
• Application Notes: This antibody is CyTOF ready.

Reactivity Notes

Mouse reactivity reported in scientific literature (PMID: 30833751).

Packaging, Storage & Formulations

• Storage: Store at -20C. Avoid freeze-thaw cycles.
• Buffer: PBS
• Preservative: No Preservative
• Concentration: 1.0 mg/ml
• Purity: Protein G purified

Alternate Names for CD47 Antibody (B6H12.2) - Azide and BSA Free

• antigen identified by monoclonal 1D8
• Antigenic surface determinant protein OA3
• CD47 antigen (Rh-related antigen, integrin-associated signal transducer)
• CD47 antigen
• CD47 glycoprotein
• CD47 molecule
• CD47
• IAP
• IAPintegrin associated protein
• Integrin-associated protein
• leukocyte surface antigen CD47
• MER6
• MER6integrin-associated signal transducer
• OA3
• Protein MER6
• Rh-related antigen

Background

CD47, originally named integrin-associated protein (IAP), is a 50-kDa protein containing five membrane spanning sequences and a short cytoplasmic tail. CD47 appears to be important in myeloid cell activation and migration across endothelial and epithelial monolayers. Anti-CD47 antibody has been shown to inhibit polymorphonuclear neutrophil (PMN) transmigration across cell monolayers and matrix. CD47 and its ligands, thrombospondin 1 (TSP-1) and SIRP-, are important regulators of dendritic cells. Recently, it has been shown that CD47 and TLR2 cross-talk to regulate PMN transmigration. The activation of TLR2/6 by MALP-2, a TLR2 specific agonist, potently inhibits human and murine PMN transmigration. The cross-talk between TLR2 and CD47 may be mediated through MYD88.

Limitations

This product is for research use only and is not approved for use in humans or in clinical diagnosis. Primary Antibodies are guaranteed for 1 year from date of receipt.

Publications for CD47 Antibody (NBP2-31106)(6)

Publications using NBP2-31106

• Singla B, Aithbathula RV, Pervaiz N et al. CD47 Activation by Thrombospondin-1 in Lymphatic Endothelial Cells Suppresses Lymphangiogenesis and Promotes Atherosclerosis Arteriosclerosis, thrombosis, and vascular biology 2023-07-01 [PMID: 37259865] (IHC-P, WB, Mouse, Human)
  
  Details:
  IHC Dilution: 1:100
• Scheepstra KWF, Mizee MR, van Scheppingen J et al. Microglia transcriptional profiling in major depressive disorder shows inhibition of cortical grey matter microglia Biological psychiatry 2023-04-28 [PMID: 37121366] (WB)
• Dai S, Liu Y, Zhao F et al. Aqueous extract of Taxus chinensis var. mairei targeting CD47 enhanced antitumor effects in non-small cell lung cancer Biomedicine & Pharmacotherapy 2022-10-01 [PMID: 36058145] (WB, IP, IHC-P, ICC/IF, Mouse, Human)
• Logtenberg MEW, Jansen JHM, Raaben M et al. Glutaminyl cyclase is an enzymatic modifier of the CD47-SIRP alpha axis and a target for cancer immunotherapy Nat. Med. 2019-04-01 [PMID: 30833751] (IP, Mouse)
• Davis RM, Kiss B, Trivedi DR el al. Surface-Enhanced Raman Scattering Nanoparticles for Multiplexed Imaging of Bladder Cancer Tissue Permeability and Molecular Phenotype ACS Nano. 2018-10-23 [PMID: 30203645] (IHC-P, Human)
• Kojima Y, Volkmer JP, McKenna K et al. CD47-blocking antibodies restore phagocytosis and prevent atherosclerosis Nature 2016-07-20 [PMID: 27437576]

Reviews for CD47 Antibody (NBP2-31106) (3)

Average Rating: 4.7

(Based on 3 reviews)

We have 3 reviews tested in 1 species: Human.

reviewed by: Verified Customer IHC-P Human 11/04/2021

Summary

• Application: Immunohistochemistry-Paraffin
• Sample Tested: Breast cancer tissue
• Species: Human
• Lot: B6H12.2

Comments

• Comments: 4% formaldehyde fixed paraffin sections.

reviewed by: Verified Customer WB Human 05/19/2019

Summary

• Application: Western Blot
• Sample Tested: 20 ug whole cell lysate
• Species: Human
• Lot: AB10610A-06

Comments

• Comments: Samples were prepared in 4X lamelli buffer and were not boiled. Boiling causes loss of signal in WB.

reviewed by: Verified Customer ICC Human 04/25/2018

Summary

• Application: Immunocytochemistry
• Sample Tested: Human trabecular meshwork
• Species: Human
• Lot: AB010610A-05

Bioinformatics

• Gene Symbol: CD47
• OMIM:
  • 601028(Human)