Anti-CD47 antibody [RM1014] (ab284132)

Rabbit Recombinant Multiclonal CD47 antibody. Suitable for ICC/IF, Flow Cyt, WB, IHC-P and reacts with Human samples. Cited in 4 publications.

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Images

Western blot - Anti-CD47 antibody [RM1014] (AB284132), expandable thumbnail

Publications

Key facts

Isotype: IgG
Host species: Rabbit
Storage buffer: pH: 7.2 - 7.4
Preservative: 0.01% Sodium azide
Constituents: 59% PBS, 40% Glycerol (glycerin, glycerine), 0.05% BSA
Form: Liquid
Clonality: Multiclonal

Immunogen

The exact immunogen used to generate this antibody is proprietary information.

Reactivity data

Select an application

Product promiseTestedExpectedPredictedNot recommended

	ICC/IF	IP	Flow Cyt	WB	IHC-P
Human	Tested	Not recommended	Tested	Tested	Tested
Mouse	Not recommended	Not recommended	Not recommended	Not recommended	Not recommended
Rat	Not recommended	Not recommended	Not recommended	Not recommended	Not recommended

Tested
Tested

Species	Dilution info	Notes
Species Human	Dilution info 1/50	Notes -

Not recommended
Not recommended

Species	Dilution info	Notes
Species Mouse	Dilution info -	Notes Perform heat-mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
Species Rat	Dilution info -	Notes -

Not recommended
Not recommended

Species	Dilution info	Notes
Species Mouse, Human, Rat	Dilution info -	Notes -

Tested
Tested

Species	Dilution info	Notes
Species Human	Dilution info 1/500	Notes -

Not recommended
Not recommended

Species	Dilution info	Notes
Species Mouse, Rat	Dilution info -	Notes -

Tested
Tested

Species	Dilution info	Notes
Species Human	Dilution info 1/1000	Notes -

Not recommended
Not recommended

Species	Dilution info	Notes
Species Mouse, Rat	Dilution info -	Notes -

Tested
Tested

Species	Dilution info	Notes
Species Human	Dilution info 1/500	Notes -

Not recommended
Not recommended

Species	Dilution info	Notes
Species Mouse, Rat	Dilution info -	Notes -

Target data

See full target information CD47

Function

Adhesive protein that mediates cell-to-cell interactions (PubMed:11509594, PubMed:15383453). Acts as a receptor for thrombospondin THBS1 and as modulator of integrin signaling through the activation of heterotrimeric G proteins (PubMed:19004835, PubMed:7691831, PubMed:8550562). Involved in signal transduction, cardiovascular homeostasis, inflammation, apoptosis, angiogenesis, cellular self-renewal, and immunoregulation (PubMed:11509594, PubMed:15383453, PubMed:19004835, PubMed:27742621, PubMed:32679764, PubMed:7691831, PubMed:8550562). Plays a role in modulating pulmonary endothelin EDN1 signaling (PubMed:27742621). Modulates nitrous oxide (NO) signaling, in response to THBS1, hence playing a role as a pressor agent, supporting blood pressure (By similarity). Plays an important role in memory formation and synaptic plasticity in the hippocampus (By similarity). Receptor for SIRPA, binding to which prevents maturation of immature dendritic cells and inhibits cytokine production by mature dendritic cells (PubMed:11509594). Interaction with SIRPG mediates cell-cell adhesion, enhances superantigen-dependent T-cell-mediated proliferation and costimulates T-cell activation (PubMed:15383453). Positively modulates FAS-dependent apoptosis in T-cells, perhaps by enhancing FAS clustering (By similarity). Plays a role in suppressing angiogenesis and may be involved in metabolic dysregulation during normal aging (PubMed:32679764). In response to THBS1, negatively modulates wound healing (By similarity). Inhibits stem cell self-renewal, in response to THBS1, probably by regulation of the stem cell transcription factors POU5F1/OCT4, SOX2, MYC/c-Myc and KLF4 (By similarity). May play a role in membrane transport and/or integrin dependent signal transduction (PubMed:7691831). May prevent premature elimination of red blood cells (By similarity).

Alternative names

CD47, MER6, Leukocyte surface antigen CD47, Antigenic surface determinant protein OA3, Integrin-associated protein, Protein MER6, IAP

Recommended products

Rabbit Recombinant Multiclonal CD47 antibody. Suitable for ICC/IF, Flow Cyt, WB, IHC-P and reacts with Human samples. Cited in 4 publications.

Key facts

Isotype: IgG
Host species: Rabbit
Storage buffer: pH: 7.2 - 7.4
Preservative: 0.01% Sodium azide
Constituents: 59% PBS, 40% Glycerol (glycerin, glycerine), 0.05% BSA
Form: Liquid
Clonality: Multiclonal
Immunogens: The exact immunogen used to generate this antibody is proprietary information.
Clone number: RM1014
Purification technique: Affinity purification Protein A
Concentration

Storage

Shipped at conditions: Blue Ice
Appropriate short-term storage duration: 1-2 weeks
Appropriate short-term storage conditions: +4°C
Appropriate long-term storage conditions: -20°C
Aliquoting information: Upon delivery aliquot
Storage information: Avoid freeze / thaw cycle

Notes

This product is a recombinant multiclonal antibody, which offers several advantages including:

- High batch-to-batch consistency and reproducibility
- Improved sensitivity and specificity
- Long-term security of supply
- Animal-free batch production

For more information, read more on recombinant antibodies.

Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMAb® patents.

Supplementary info

This supplementary information is collated from multiple sources and compiled automatically.

Activity summary

CD47 also referred to as integrin-associated protein carries a molecular weight of approximately 50 kDa. This protein is broadly expressed across various cell types notably on erythrocytes leukocytes and endothelial cells. Its transmembrane glycoprotein structure allows it to perform various cellular functions. CD47 interacts with specific ligands prominently SIRPα which are present on immune cells such as macrophages and dendritic cells. The engagement of CD47 with these ligands plays a major role in cellular signaling processes.

Biological function summary

CD47 functions as a "don't eat me" signal inhibiting phagocytosis by binding to SIRPα on macrophages. This protein is also important for cell adhesion and migration and can modulate interactions with integrins. While not a part of a large complex CD47 associates with various cytoskeletal and membrane proteins to maintain cellular architecture and transmission of mechanical forces. Additionally its interaction with thrombospondins influences angiogenesis and nitric oxide signaling.

Pathways

CD47 is extensively involved in the regulation of the immune response and angiogenesis. The interaction between CD47 and SIRPα plays a part in the regulation of macrophage activity impacting the immune signaling pathway. CD47 also interacts with integrins allowing it to contribute to the angiogenesis pathway which is critical in the formation of new blood vessels. These interactions help coordinate complex cellular responses and maintain tissue homeostasis.

Associated diseases and disorders

CD47 levels have associations with cancer and chronic inflammatory diseases. In cancer CD47 overexpression can allow tumor cells to evade the immune response by downregulating phagocytosis through SIRPα interaction. Therapeutically targeting CD47 using agents like anti-CD47 antibodies shows potential for enhancing anti-tumor immunity. Additionally CD47 is involved in atherosclerosis where its interaction with thrombospondin-1 contributes to disease pathogenesis by affecting nitric oxide signaling and vascular remodeling.

Product promise

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In the unlikely event of one of our products not working as expected, you are covered by our product promise.

Full details and terms and conditions can be found here:
Terms & Conditions.

11 product images

Western blot - Anti-CD47 antibody [RM1014] (ab284132)
Blocking and diluting buffer and concentration was 5% NFDM/TBST.
Low expression control: HepG2 (PMID: 25721088, 30415063).
We recommend that samples are not boiled after adding loading buffer as this may cause protein aggregates.
The molecular mass observed is consistent with the literature (PMID 12393467, PMID 11034562).
All lanes: Western blot - Anti-CD47 antibody [RM1014] (ab284132) at 1/1000 dilution
Lane 1: Jurkat (Human T cell leukemia T lymphocyte) whole cell lysate 20ug
Lane 2: U-937 (Human histiocytic lymphoma monocyte) whole cell lysate 20ug
Lane 3: HepG2 (Human hepatocellular carcinoma epithelial cell) whole cell lysate 20ug
Secondary
All lanes: Western blot - Goat Anti-Rabbit IgG H&L (HRP) (Goat Anti-Rabbit IgG H&L (HRP) ab97051) at 1/100000 dilution
Exposure time: 37s
Western blot - Anti-CD47 antibody [RM1014] (ab284132)
Blocking and diluting buffer and concentration was 5% NFDM/TBST.
We recommend that samples are not boiled after adding loading buffer as this may cause protein aggregates.
All lanes: Western blot - Anti-CD47 antibody [RM1014] (ab284132) at 1/1000 dilution
Lane 1: Human liver lysate at 20 µg
Lane 2: Human ovary cancer lysate at 20 µg
Secondary
All lanes: Western blot - Goat Anti-Rabbit IgG H&L (HRP) (Goat Anti-Rabbit IgG H&L (HRP) ab97051) at 1/100000 dilution
Exposure time: 37s
Western blot - Anti-CD47 antibody [RM1014] (ab284132)
Blocking and diluting buffer and concentration was 5% NFDM/TBST.
We recommend that samples are not boiled after adding loading buffer as this may cause protein aggregates.
All lanes: Western blot - Anti-CD47 antibody [RM1014] (ab284132) at 1/1000 dilution
Lane 1: Wild-type HEK-293T (Human embryonic kidney epithelial cell, ab255449) whole cell lysate at 20 µg
Lane 2: CD47 knockout HEK-293T (Human CD47 knockout HEK-293T cell line ab266324) whole cell lysate at 20 µg
Secondary
All lanes: Western blot - Goat Anti-Rabbit IgG H&L (HRP) (Goat Anti-Rabbit IgG H&L (HRP) ab97051) at 1/100000 dilution
Exposure time: 37s
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD47 antibody [RM1014] (ab284132)
Immunohistochemical analysis of paraffin-embedded human spleen tissue labelling CD47 with ab284132 at 1/500 followed by a ready to use LeicaDS9800 (BOND™ Polymer Refine Detection). Positive staining on human spleen. The section was incubated with ab284132 for 30 mins at room temperature. The immunostaining was performed on a Leica Biosystems BOND^® RX instrument. Counterstained with Hematoxylin.
Secondary antibody only control: Secondary antibody is a ready to use LeicaDS9800 (BOND™ Polymer Refine Detection).
Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution2) for 20 mins.
Flow Cytometry - Anti-CD47 antibody [RM1014] (ab284132)
Flow cytometric analysis of HepG2 (Human hepatocellular carcinoma epithelial cell, Left) / U-937 (Human histiocytic lymphoma monocyte, Right) cells labelling CD47 with ab284132 at 1/500 dilution (0.1μg) (Red) compared with a Rabbit monoclonal IgG (Rabbit IgG, monoclonal [EPR25A] - Isotype Control ab172730) (Black) isotype control and an unlabelled control (cells without incubation with primary antibody and secondary antibody) (Blue). Goat Anti-Rabbit IgG (Alexa Fluor^® 488, Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed ab150081) at 1/2000 dilution was used as the secondary antibody. Low expression control: HepG2 (PMID: 25721088, 30415063). Gated on viable cells.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD47 antibody [RM1014] (ab284132)
Immunohistochemical analysis of paraffin-embedded human placenta tissue labelling CD47 with ab284132 at 1/500 followed by a ready to use LeicaDS9800 (BOND™ Polymer Refine Detection). Positive staining on human placenta. The section was incubated with ab284132 for 30 mins at room temperature.The immunostaining was performed on a Leica Biosystems BOND^® RX instrument. Counterstained with Hematoxylin.
Secondary antibody only control: Secondary antibody is a ready to use LeicaDS9800 (BOND™ Polymer Refine Detection).
Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution2) for 20 mins.
Flow Cytometry - Anti-CD47 antibody [RM1014] (ab284132)
Flow cytometric analysis of HepG2 (Human hepatocellular carcinoma epithelial cell, Left) / Jurkat (Human T cell leukemia T lymphocyte, Right) cells labelling CD47 with ab284132 at 1/500 dilution (0.1μg) (Red) compared with a Rabbit monoclonal IgG (Rabbit IgG, monoclonal [EPR25A] - Isotype Control ab172730) (Black) isotype control and an unlabelled control (cells without incubation with primary antibody and secondary antibody) (Blue). Goat Anti-Rabbit IgG (Alexa Fluor^® 488, Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed ab150081) at 1/2000 dilution was used as the secondary antibody. Low expression control: HepG2 (PMID: 25721088, 30415063). Gated on viable cells.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD47 antibody [RM1014] (ab284132)
Immunohistochemical analysis of paraffin-embedded human breast cancer tissue labelling CD47 with ab284132 at 1/500 followed by a ready to use LeicaDS9800 (BOND™Polymer Refine Detection). Positive staining on human breast cancer (PMID: 31528120). The section was incubated with ab284132 for 30 mins at room temperature.The immunostaining was performed on a Leica Biosystems BOND^® RX instrument. Counterstained with Hematoxylin.
Secondary antibody only control: Secondary antibody is a ready to use LeicaDS9800 (BOND™ Polymer Refine Detection).
Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution2) for 20 mins.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD47 antibody [RM1014] (ab284132)
Immunohistochemical analysis of paraffin-embedded human ovarian carcinoma tissue labelling CD47 with ab284132 at 1/500 (0.924 μg/ml) followed by a ready to use LeicaDS9800 (BOND™ Polymer Refine Detection) was used. Positive staining on human ovarian carcinoma. The section was incubated with ab284132 for 30 mins at room temperature.The immunostaining was performed on a Leica Biosystems BOND^® RX instrument. Counterstained with Hematoxylin.
Secondary antibody only control: Secondary antibody is a ready to use LeicaDS9800 (BOND™ Polymer Refine Detection).
Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution2) for 20 mins.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD47 antibody [RM1014] (ab284132)
Immunohistochemical analysis of paraffin-embedded human ovarian carcin tissue labelling CD47 with ab284132 at 1/500 followed by a ready to use LeicaDS9800 (BOND™Polymer Refine Detection) was used. Positive staining on human ovarian carcinoma. The section was incubated with ab284132 for 30 mins at room temperature.The immunostaining was performed on a Leica Biosystems BOND^® RX instrument. Counterstained with Hematoxylin.
Secondary antibody only control: Secondary antibody is a ready to use LeicaDS9800 (BOND™ Polymer Refine Detection).
Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution2) for 20 mins.
Immunocytochemistry/ Immunofluorescence - Anti-CD47 antibody [RM1014] (ab284132)
Immunofluorescent analysis of 100% methanol-fixed, 0.1% Triton X-100 permeabilized Jurkat cells labelling CD47 with ab284132 at 1/50 dilution, followed by Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor^® 488) preadsorbed antibody at 1/1000 dilution (Green). Confocal image showing membranous staining in Jurkat cell line. Negative control: HepG2 (PMID: 25721088). Alexa Fluor® 594 Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker ab195889 Anti-alpha Tubulin mouse monoclonal antibody - Microtubule Marker (Alexa Fluor^® 594) was used to counterstain tubulin at 1/200 dilution (Red). The Nuclear counterstain was DAPI (Blue).
Secondary antibody only control: Secondary antibody is Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor^® 488) preadsorbed at 1/1000 dilution.