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

Novus Biologicals | Catalog # NBP2-31106

Novus Biologicals
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Key Product Details

Species Reactivity

Validated:

Human, Mouse

Cited:

Human, Mouse

Applications

Validated:

Immunohistochemistry, Immunohistochemistry-Paraffin, Western Blot, Flow Cytometry, Flow (Cell Surface), Immunocytochemistry/ Immunofluorescence, Immunoprecipitation, CyTOF-ready

Cited:

Immunohistochemistry-Paraffin, Western Blot, Immunocytochemistry/ Immunofluorescence, Immunoprecipitation

Label

Unconjugated

Antibody Source

Monoclonal Mouse IgG1 Clone # B6H12.2

Format

Azide and BSA Free
View all CD47 Primary Antibodies »

Product Specifications

Immunogen

This antibody was raised against CD47.

Reactivity Notes

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

Clonality

Monoclonal

Host

Mouse

Isotype

IgG1

Scientific Data Images for CD47 Antibody (B6H12.2) - Azide and BSA Free

Immunocytochemistry/ Immunofluorescence: CD47 Antibody (B6H12.2) - Azide and BSA Free [NBP2-31106]

Immunocytochemistry/ Immunofluorescence: CD47 Antibody (B6H12.2) - Azide and BSA Free [NBP2-31106]

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 and BSA Free [NBP2-31106]

Western Blot: CD47 Antibody (B6H12.2)Azide and BSA Free [NBP2-31106]

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 and BSA Free [NBP2-31106]

Immunohistochemistry-Paraffin: CD47 Antibody (B6H12.2) - Azide and BSA Free [NBP2-31106]

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 and BSA Free [NBP2-31106]

Flow Cytometry: CD47 Antibody (B6H12.2) - Azide and BSA Free [NBP2-31106]

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 and BSA Free [NBP2-31106]

Immunocytochemistry: CD47 Antibody (B6H12.2) - Azide and BSA Free [NBP2-31106]

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 and BSA Free [NBP2-31106]

Immunocytochemistry/ Immunofluorescence: CD47 Antibody (B6H12.2) - Azide and BSA Free [NBP2-31106]

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 and BSA Free [NBP2-31106]

Immunohistochemistry-Paraffin: CD47 Antibody (B6H12.2) - Azide and BSA Free [NBP2-31106]

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 and BSA Free [NBP2-31106]

Immunohistochemistry-Paraffin: CD47 Antibody (B6H12.2) - Azide and BSA Free [NBP2-31106]

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 and BSA Free [NBP2-31106]

Flow (Cell Surface): CD47 Antibody (B6H12.2) - Azide and BSA Free [NBP2-31106]

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].

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

Application
Recommended Usage

Flow Cytometry

1 ug/mL

Immunohistochemistry-Paraffin

0.5 ug/mL

Western Blot

1:1000
Application Notes
This antibody is CyTOF ready.

Reviewed Applications

Read 5 reviews rated 4.8 using NBP2-31106 in the following applications:

Flow Cytometry Panel Builder

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Flow Cytometry

Formulation, Preparation, and Storage

Purification

Protein G purified

Formulation

PBS

Format

Azide and BSA Free

Preservative

No Preservative

Concentration

1.0 mg/ml

Shipping

The product is shipped with polar packs. Upon receipt, store it immediately at the temperature recommended below.

Stability & Storage

Store at -20C. Avoid freeze-thaw cycles.

Background: CD47

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.

Alternate Names

CD47, IAP, MER6, OA3

Gene Symbol

CD47

OMIM

601028 (Human)

Additional CD47 Products

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

Certificate of Analysis

To download a Certificate of Analysis, please enter a lot or batch number in the search box below.

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Product Specific Notices for CD47 Antibody (B6H12.2) - Azide and BSA Free

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.

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

Customer Reviews for CD47 Antibody (B6H12.2) - Azide and BSA Free (5)

4.8 out of 5
5 Customer Ratings
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4 Stars
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Customer Images

Showing  1 - 5 of 5 reviews Showing All
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  • MCF7 treatment
    Name: Anonymous
    Application: Cell culture
    Sample Tested: MCF-7 human breast cancer cell line
    Species: Human
    Verified Customer | Posted 05/29/2025
    MCF7 treated with CD47 antibody
    MCF7 cells were seeded on 96well plate with 10 ug/mL of CD47 and imaged after 24h
    CD47 Antibody (B6H12.2) - Azide and BSA Free NBP2-31106
    Bio-Techne Response
    This review reflects a new species or application tested on a primary antibody.
  • HCT116 treatment
    Name: Anonymous
    Application: Cell Culture treatment
    Sample Tested: HCT-116 human colorectal carcinoma cell line
    Species: Human
    Verified Customer | Posted 05/26/2025
    5000 cells were treated with 10 ug/mL of CD47 or IgG1 isotope control
    Cell culture treatment - 5000 cells were treated with 10 ug/mL of antibody
    CD47 Antibody (B6H12.2) - Azide and BSA Free NBP2-31106
    Bio-Techne Response
    This review reflects a new species or application tested on a primary antibody.
  • CD47 Antibody (B6H12.2) - Azide Free
    Name: Anonymous
    Application: Immunohistochemistry-Paraffin
    Sample Tested: Breast cancer tissue
    Species: Human
    Verified Customer | Posted 11/04/2021
    Breast cancer tissue
    4% formaldehyde fixed paraffin sections.
    CD47 Antibody (B6H12.2) - Azide and BSA Free NBP2-31106
  • CD47 Antibody (B6H12.2) - Azide Free
    Name: Anonymous
    Application: Western Blot
    Sample Tested: 20 ug whole cell lysate
    Species: Human
    Verified Customer | Posted 05/19/2019
    Detection of CD47 expression on two hematological cancer cell lines, Jurkat and Raji.
    Samples were prepared in 4X lamelli buffer and were not boiled. Boiling causes loss of signal in WB.
    CD47 Antibody (B6H12.2) - Azide and BSA Free NBP2-31106
  • CD47 Antibody (B6H12.2) - Azide Free
    Name: Anonymous
    Application: Immunocytochemistry
    Sample Tested: Human trabecular meshwork
    Species: Human
    Verified Customer | Posted 04/25/2018
    CD47 Antibody (B6H12.2) - Azide and BSA Free NBP2-31106

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Protocols

View specific protocols for CD47 Antibody (B6H12.2) - Azide and BSA Free (NBP2-31106):

Protocol for Flow Cytometry Cell Surface Staining
Sample Preparation.
1. Grow cells to 60-85% confluency. Flow cytometry requires between 2 x 105 and 1 x 106 cells for optimal performance.
2. If cells are adherent, harvest gently by washing once with staining buffer and then scraping. Avoid using trypsin as this can disrupt certain epitopes of interest. If enzymatic harvest is required, use Accutase, Collagenase, or TrypLE Express for a less damaging option.
3. Reserve 100 uL for counting, then transfer cell volume into a 15 mL conical tube and centrifuge for 4 minutes at 400 RCF.
a. Count cells using a hemocytometer and a 1:1 trypan blue exclusion stain to determine cell viability before starting the flow protocol. If cells appear blue, do not proceed.
4. Re-suspend cells to a concentration of 1 x 106 cells/mL in staining buffer (NBP2-26247).
5. Aliquot out 100 uL samples in accordance with your experimental samples.

Tip: When cell surface and intracellular staining are required in the same sample, it is advisable that the cell surface staining be performed first since the fixation and permeablization steps might reduce the availability of surface antigens.
Cell surface staining
1. Recommended: Block non-specific interactions using 0.5-1 ug of a species specific Fc-blocking reagent such as an anti-mouse CD16/CD32 antibody (NBP1-27946).
2. Add appropriate amount of each antibody (eg. 1 test or 1 ug per sample, as experimentally determined) to 100 uL of staining buffer (NBP2-26247) per sample (eg. use 1 mL of staining buffer for 10 samples).
3. Mix well and incubate at room temperature in dark for 20 minutes.
4. Add 1-2 mL of staining buffer and centrifuge at 400 RCF for 1 minute and discard supernatant.
5. Wash twice by re-suspending cells in staining buffer (2 mL for tubes or 200 uL for wells) and centrifuging at 400 RCF for 5 minutes. Discard supernatant.
6. Add appropriate amount of secondary antibody (as experimentally determined) to each sample.
7. Incubate at room temperature in dark for 20 minutes.
8. Add 1-2 mL of staining buffer and centrifuge at 400 RCF for 1 minute and discard supernatant.
9. Wash twice by re-suspending cells in staining buffer (2 mL for tubes or 200 uL for wells) and centrifuging at 400 RCF for 5 minutes. Discard supernatant.
10. Resuspend in an appropriate volume of staining buffer (usually 500 uL per sample) and proceed with analysis on your flow cytometer.

Immunocytochemistry Protocol

Culture cells to appropriate density in 35 mm culture dishes or 6-well plates.

1. Remove culture medium and wash the cells briefly in PBS. Add 10% formalin to the dish and fix at room temperature for 10 minutes.
2. Remove the formalin and wash the cells in PBS.
3. Permeablize the cells with 0.1% Triton X100 or other suitable detergent for 10 min.
4. Remove the permeablization buffer and wash three times for 10 minutes each in PBS. Be sure to not let the specimen dry out.
5. To block nonspecific antibody binding, incubate in 10% normal goat serum from 1 hour to overnight at room temperature.
6. Add primary antibody at appropriate dilution and incubate overnight at 4C.
7. Remove primary antibody and replace with PBS. Wash three times for 10 minutes each.
8. Add secondary antibody at appropriate dilution. Incubate for 1 hour at room temperature.
9. Remove secondary antibody and replace with PBS. Wash three times for 10 minutes each.
10. Counter stain DNA with DAPi if required.

Immunohistochemistry-Paraffin Embedded Sections

Antigen Unmasking:
Bring slides to a boil in 10 mM sodium citrate buffer (pH 6.0) then maintain at a sub-boiling temperature for 10 minutes. Cool slides on bench-top for 30 minutes (keep slides in the sodium citrate buffer all the time).

Staining:
1. Wash sections in deionized water three times for 5 minutes each.
2. Wash sections in PBS for 5 minutes.
3. Block each section with 100-400 ul blocking solution (1% BSA in PBS) for 1 hour at room temperature.
4. Remove blocking solution and add 100-400 ul diluted primary antibody. Incubate overnight at 4 C.
5. Remove antibody solution and wash sections in wash buffer three times for 5 minutes each.
6. Add 100-400 ul HRP polymer conjugated secondary antibody. Incubate 30 minutes at room temperature.
7. Wash sections three times in wash buffer for 5 minutes each.
8. Add 100-400 ul DAB substrate to each section and monitor staining closely.
9. As soon as the sections develop, immerse slides in deionized water.
10. Counterstain sections in hematoxylin.
11. Wash sections in deionized water two times for 5 minutes each.
12. Dehydrate sections.
13. Mount coverslips.

Western Blot Protocol

1. Perform SDS-PAGE on samples to be analyzed, loading 10-25 ug of total protein per lane.
2. Transfer proteins to PVDF membrane according to the instructions provided by the manufacturer of the membrane and transfer apparatus.
3. Stain the membrane with Ponceau S (or similar product) to assess transfer success, and mark molecular weight standards where appropriate.
4. Rinse the blot TBS -0.05% Tween 20 (TBST).
5. Block the membrane in 5% Non-fat milk in TBST (blocking buffer) for at least 1 hour.
6. Wash the membrane in TBST three times for 10 minutes each.
7. Dilute primary antibody in blocking buffer and incubate overnight at 4C with gentle rocking.
8. Wash the membrane in TBST three times for 10 minutes each.
9. Incubate the membrane in diluted HRP conjugated secondary antibody in blocking buffer (as per manufacturer's instructions) for 1 hour at room temperature.
10. Wash the blot in TBST three times for 10 minutes each (this step can be repeated as required to reduce background).
11. Apply the detection reagent of choice in accordance with the manufacturers instructions.

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Associated Pathways

A-beta Pathways: Uptake & Degradation A-beta Pathways: Uptake & Degradation Thumbnail