CD172a antibody | 15-414

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Mouse anti Human CD172a:StarBright UltraViolet 700

Product Type: Monoclonal Antibody
Clone: 15-414
Isotype: IgG2a
Specificity: CD172a

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MCA2518SBUV700 Datasheet SDS SDS	F *	100 Tests/0.5ml		Log in
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Anti Human CD172a Antibody, clone 15-414 thumbnail image 1

Anti Human CD172a Antibody, clone 15-414 thumbnail image 2

Anti Human CD172a Antibody, clone 15-414 thumbnail image 3

Anti Human CD172a Antibody, clone 15-414 thumbnail image 4

Anti Human CD172a Antibody, clone 15-414 thumbnail image 5

Anti Human CD172a Antibody, clone 15-414 thumbnail image 6

Anti Human CD172a Antibody, clone 15-414 thumbnail image 7

Anti Human CD172a Antibody, clone 15-414 gallery image 1 — Flow cytometry analysis of human peripheral blood monocytes stained with RPE conjugated Mouse anti Human CD172a antibody, clone 15-414 (**MCA2518PE**).

Anti Human CD172a Antibody, clone 15-414 gallery image 2 — **Published customer image:**
Mouse anti Human CD172 antibody, clone 15-414 (**MCA2518**) used for the demonstration of SIRPα expression on human thymus cells by immunofluorescence and flow cytometry.
**Image caption:**
Thymic Sirpα⁺DC-SIGN⁺ cDCs can uptake antigens, which leak inside the IVRs.
(A) Triple-color fluorescent image with the combination of Col IV (green), Ly-51 (blue), and Sirpα (red) in mouse thymic tissue. Dashed lines indicate the boundary between cortex (C) and medulla (M). Scale bar, 100 μm. (B) A double-color fluorescent immunostaining for DC-SIGN (green) and Sirpα (red) in human thymic tissue. Dashed lines indicate the outer margin of thymic lobule. Scale bar, 100 μm. (C) Thymic B220⁻CD11c^highSirpα⁻ (Sirpα⁻) and B220⁻CD11c^highSirpα⁺ (Sirpα⁺) cDCs were sorted (Sirpα⁻ purity, 97.8%; Sirpα⁺ purity, 97.1%) by using FACSAria (BD Biosciences), from thymic low-density cells isolated from 10 mice. Expression of DC-SIGN, SIGNR1, SIGNR2, SIGNR3, and SIGNR4 was determined by RT-PCR. GAPDH served as an internal positive control. RT-PCR analysis was repeated three times. (D) DC-SIGN expression on CD11c^highSirpα⁻ and CD11c^highSirpα⁺ cDCs. Gray-filled and red-open histograms indicate the results obtained using isotype control and specific mAbs for DC-SIGN, respectively. (E) Thymi were collected at the indicated time points after FITC-conjugated dextran was intravenously injected into WT mice. Triple-color fluorescent images with the combination of dextran (green), DAPI (blue), and Col IV (red) are shown here. Scale bar, 200 μm. (F) Triple-color fluorescent images with the combination of dextran (green), Col IV (blue), and Sirpα (red) at 6 hrs after dextran injection are shown here. The confocal images in the parenchyma nearby small vessel and inside the IVRs, and superposed 3-D image inside the IVRs are shown in the upper and lower left, and lower right panels, respectively. Scale bars, 50 μm. (G) Uptake of OVA647 in low density cells at 4 hrs after injection. Percentage of CD11c (+) OVA647 (+) and CD11c (−) OVA647 (+) region are shown. Expression of Sirp&apha; molecule in region 1 (R1) and R2 are represented by unfilled and gray-filled histogram, respectively. (H) Uptake of OVA647 at 4 hrs after intravenous, subcutaneous, or intraperitoneal injection. Percentage of Sirp&apha; (+) OVA647 (+) region in thymic CD11c^high DCs is shown in each panel. PBS was injected as a control. Representative results from three independent experiments are shown here.

From: Baba T, Badr MES, Tomaru U, Ishizu A, Mukaida N (2012)
Novel Process of Intrathymic Tumor-Immune Tolerance through CCR2-Mediated Recruitment of Sirpα⁺ Dendritic Cells: A Murine Model.
**PLoS ONE 7(7): e41154**.
doi: 10.1371/journal.pone.0041154.
This image is from an open access article distributed under the terms of the Creative Commons Attribution License.

Anti Human CD172a Antibody, clone 15-414 gallery image 3 — **Published customer image:**
Mouse anti Human CD172 antibody, clone 15-414 (**MCA2518**) used for the demonstration of SIRPα expression on human thymus cells by flow cytometry.
**Image caption:**
Mice deficient in CCR2 gene exhibited reduced Treg differentiation and negative selection in the thymus.(A) Schematic representation of localization and function of thymic Sirpα⁺ cDCs. OVA protein uptake by CD11c^highSirpα⁺ cDCs in WT and CCR2^−/− thymus at 4 hrs after injection are shown. Percentage of Sirpα (+) OVA647 (−) and Sirpα (+) OVA647 (+) region are shown in each panel. (B) The uptake of OVA647 by CD11c⁺Sirpα⁺ cDCs derived from C57BL/6 thymi at 4 hrs after injection is shown in left panel. Sirp&apha;⁺ cDCs capturing OVA647 were separated into two groups according to the efficiency of OVA647 uptake; low and high. Percentages of cells in low and high regions in WT and CCR2^−/− mice in BALB/c and C57BL/6 strain are shown in the right panel. Data represent mean ± SD from five and three independent experiments in BALB/c and C57BL/6 mice, respectively. (C) Expression of CD25 and Foxp3 on DO11.10^high (R1) thymocytes at 2 days after OVA protein injection. Percentage of Foxp3 (+) region is shown in each panel. Representative results from three independent experiments are shown. (D) The numbers of Foxp3⁺ mature thymocytes were determined on DO11.10 and DO11.10/CCR2^−/− mice at 2 days after OVA injection. Each symbol represents an individual mouse. Small horizontal lines indicate the mean. (E) Clonal deletion of thymocytes after OVA injection. Two mg OVA protein or heat-aggregated OVA protein were intravenously injected into DO11.10 mice. At 2 days after injection, the numbers of total thymocytes (left graph) and DP thymocytes (right graph) were determined on DO11.10 and DO11.10/CCR2^−/− mice. Each symbol represents an individual mouse. **, p<0.01. *, p<0.05. N.S., no significant difference.

From: Baba T, Badr MES, Tomaru U, Ishizu A, Mukaida N (2012)
Novel Process of Intrathymic Tumor-Immune Tolerance through CCR2-Mediated Recruitment of Sirpα⁺ Dendritic Cells: A Murine Model.
**PLoS ONE 7(7): e41154**.
doi: 10.1371/journal.pone.0041154.
This image is from an open access article distributed under the terms of the Creative Commons Attribution License.

Anti Human CD172a Antibody, clone 15-414 gallery image 4 — Human whole blood was stained with Anti-Human CD16 FITC (**MCA5665F**) only (Figure A) or with Anti-Human CD16 FITC (**MCA5665F**) and Anti-Human CD172a StarBright UltraViolet 540 (**MCA2518SBUV540**) (Figure B). Experiments were performed on isolated red-cell lysed blood, gated on live, single mononuclear cells. Data were acquired on a Cytek Aurora.

Anti Human CD172a Antibody, clone 15-414 gallery image 5 — Human whole blood was stained with Anti-Human CD16 FITC (**MCA5665F**) only (Figure A) or with Anti-Human CD16 FITC (**MCA5665F**) and Anti-Human CD172a StarBright UltraViolet 700 (**MCA2518SBUV700**) (Figure B). Experiments were performed on isolated red-cell lysed blood, gated on live, single mononuclear cells. Data were acquired on a Cytek Aurora.

Anti Human CD172a Antibody, clone 15-414 gallery image 6 — Human whole blood was stained with Anti-Human CD16 FITC (**MCA5665F**) only (Figure A) or with Anti-Human CD16 FITC (**MCA5665F**) and Anti-Human CD172a StarBright UltraViolet 770 (**MCA2518SBUV770**) (Figure B). Experiments were performed on isolated red-cell lysed blood, gated on live, single mononuclear cells. Data were acquired on a Cytek Aurora.

Anti Human CD172a Antibody, clone 15-414 gallery image 7 — Human whole blood was stained with Anti-Human CD16 FITC (**MCA5665F**) only (Figure A) or with Anti-Human CD16 FITC (**MCA5665F**) and Anti-Human CD172a StarBright Violet 535 (**MCA2518SBV535**) (Figure B). Experiments were performed on isolated red-cell lysed blood, gated on live, single mononuclear cells. Data were acquired on a Cytek Aurora.

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Mouse anti Human CD172a antibody, clone 15-414 recognizes human CD172a, also known as signal-regulatory protein alpha, a receptor-type transmembrane glycoprotein expressed on cells of myeloid origin, including granulocytes, dendritic cells (DCs), macrophages, mast cells and haematopoietic stem cells.

CD172a acts as a substrate for several activated tyrosine kinases, including EGFR, PDGFR, src and insulin receptor and is involved in the negative regulation of receptor tyrosine kinase-coupled signaling pathways. Ligand binding of CD172a to integrin-associated protein CD47, results in tyrosine kinase phosphorylation of immunoreceptor tyrosine-based inhibitory motifs (ITIMs) within the cytoplasmic region of CD172a, mediating the recruitment and activation of the tyrosine phosphatases SHP-1 and SHP-2. These then act as regulators of cellular function, through dephosphorylation of specific substrates. Ligation of CD172a with CD47 has been demonstrated in several regulatory processes, including the inhibition of host cell phagocytosis by macrophages and the bi-directional activation of T cells and DCs.

Target Species: Human
Product Form: Purified IgG conjugated to StarBright UltraViolet 700 - liquid
Preparation: Purified IgG prepared by affinity chromatography on Protein A from tissue culture supernatant
Buffer Solution: Phosphate buffered saline
Preservative Stabilisers: 0.09% Sodium Azide (NaN₃)
1% Bovine Serum Albumin
0.1% Pluronic F68
0.1% PEG 3350
0.05% Tween 20
Immunogen: Monocyte-derived dendritic cells.
Approx. Protein Concentrations: For information on the concentration of our StarBright Dye conjugated reagents please visit our FAQ page.
Fusion Partners: Spleen cells from immunized Balb/c mice were fused with cells of the X63-Ag8.653 myeloma cell line.
Max Ex/Em: Fluorophore Excitation Max (nm) Emission Max (nm)

StarBright UltraViolet 700 340 700
Regulatory: For research purposes only
Guarantee: 12 months from date of despatch
Acknowledgements: This product is covered by U.S. Patent No. 10,150,841 and related U.S. and foreign counterparts

Store at +4°C. DO NOT FREEZE.
This product should be stored undiluted.

This product has been reported to work in the following applications. This information is derived from testing within our laboratories, peer-reviewed publications or personal communications from the originators. Please refer to references indicated for further information. For general protocol recommendations, please visit the antibody protocols page.

Application Name	Verified	Min Dilution	Max Dilution
Flow Cytometry ¹		Neat

¹Staining at lower temperatures generally requires longer incubation to reach antibody binding equilibrium, and this effect is more pronounced for StarBright Dyes due to their larger size. For optimum staining, Bio-Rad recommends incubating samples either overnight at 4°C or 1 hour at room temperature.

Where this product has not been tested for use in a particular technique this does not necessarily exclude its use in such procedures. Suggested working dilutions are given as a guide only. It is recommended that the user titrates the product for use in their own system using appropriate negative/positive controls.

Flow Cytometry: Use 5μl of the suggested working dilution to label 0.5x10⁶ cells in 100μl. Best practices suggest a 5 min centrifugation at 6,000g prior to sample application.

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References for CD172a antibody

Baba, T. et al. (2012) Novel Process of Intrathymic Tumor-Immune Tolerance through CCR2-Mediated Recruitment of Sirpα(+) Dendritic Cells: A Murine Model.
PLoS One. 7: e41154.
Szaraz, P. et al. (2016) In Vitro Differentiation of First Trimester Human Umbilical Cord Perivascular Cells into Contracting Cardiomyocyte-Like Cells.
Stem Cells Int. 2016: 7513252.