Do you have a protocol for the isolation and treatment of dendritic cells used for IC flow staining with Human GM-CSF Antibody, catalog #s MAB615 and IC615?

Name: Human GM-CSF PE-conjugated Antibody
Brand: R&D Systems
SKU: IC615P
Price: 379 USD

MagCellect Human CD14+ Cell Isolation Kit, catalog # MAGH105, is used to isolate CD14+ monocytes from PBMCs, as the starting population. Monocytes are cultured in media with rhIL-4 (catalog # 204-IL) at 20 ng/ml and rhGM-CSF (catalog # 215-GM) at 50ng/ml. On day 6, add LPS at 20ng/ml, rhTNF-a (catalog # 210-TA) at 20ng/ml and rhIL-1b (catalog # 201-LB) at 20ng/ml to culture medium and incubate cells overnight. Collect cells for flow cytometric analysis on day 7.

Human GM-CSF PE-conjugated Antibody

Catalog #: IC615P
2 Citations Datasheet / COA / SDS

Catalog #	Availability	Size / Price	Qty
IC615P

Detection of GM‑CSF in Human Dendritic Cells by Flow Cytometry.

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All GM-CSF Antibodies

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Citations (2)

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Summary Data Examples Preparation and Storage Reconstitution Calculator Background Product Datasheets Related Research Areas

Human GM-CSF PE-conjugated Antibody Summary

Species Reactivity

Human

Specificity

Detects human GM-CSF in ELISAs. In ELISAs, this antibody does not cross-react with recombinant mouse (rm) GM‑CSF, rrGM‑CSF, rhM‑CSF, rhG‑CSF, rhIL‑5‑R alpha, or rhIL‑3 R alpha.

Source

Monoclonal Mouse IgG₁ Clone # 6804

Purification

Protein A or G purified from hybridoma culture supernatant

Immunogen

E. coli-derived recombinant human GM-CSF
Ala18-Glu144
Accession # P04141

Formulation

Supplied in a saline solution containing BSA and Sodium Azide.

Label

Phycoerythrin (Excitation= 488 nm, Emission= 565-605 nm)

Applications

Recommended Concentration

Sample

Intracellular Staining by Flow Cytometry

10 µL/10⁶ cells

See below

Please Note: Optimal dilutions should be determined by each laboratory for each application. General Protocols are available in the Technical Information section on our website.

Scientific Data

Intracellular Staining by Flow Cytometry

Detection of GM-CSF antibody in Human Dendritic Cells antibody by Flow Cytometry.

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Detection of GM‑CSF in Human Dendritic Cells by Flow Cytometry. Human dendritic cells treated with LPS, Recombinant Human TNF-a (Catalog # 210-TA), and Recombinant Human TGF-beta 1 (Catalog # 240-B) were stained with Mouse Anti-Human GM-CSF PE-conjugated Monoclonal Antibody (Catalog # IC615P, filled histogram) or isotype control antibody (Catalog # IC002P, open histogram). To facilitate intracellular staining, cells were fixed with Flow Cytometry Fixation Buffer (Catalog # FC004) and permeabilized with Flow Cytometry Permeabilization/Wash Buffer I (Catalog # FC005). View our protocol for Staining Intracellular Molecules.

Reconstitution Calculator

Preparation and Storage

Shipping

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

Stability & Storage

Protect from light. Do not freeze.

12 months from date of receipt, 2 to 8 °C as supplied.

Background: GM-CSF

GM-CSF is a growth factor produced by a variety of lymphoid cells. It binds to receptor heterodimers consisting of a GM-CSF-specific alpha chain and the common beta chain that is shared by the high affinity receptors for IL-3 and IL-5.

Long Name

Granulocyte Macrophage Growth Factor

Entrez Gene IDs

1437 (Human); 12981 (Mouse); 116630 (Rat); 397208 (Porcine); 403923 (Canine); 493805 (Feline)

Alternate Names

colony stimulating factor 2 (granulocyte-macrophage); Colony-stimulating factor; CSF; CSF2; CSF-2; GMCSF; GM-CSF; GMCSFgranulocyte-macrophage colony-stimulating factor; granulocyte-macrophage colony stimulating factor; MGC131935; MGC138897; Molgramostim; molgramostin; Sargramostim

Product Datasheets

Product Datasheet

COA

SDS

Related Research Areas

Citations for Human GM-CSF PE-conjugated Antibody

R&D Systems personnel manually curate a database that contains references using R&D Systems products. The data collected includes not only links to publications in PubMed, but also provides information about sample types, species, and experimental conditions.

2 Citations: Showing 1 - 2
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Human CD4(+) T cells recognize an epitope within alpha-fetoprotein sequence and develop into TGF-beta-producing CD4(+) T cells.
Authors: Alisa A, Boswell S, Pathan AA, Ayaru L, Williams R, Behboudi S
J. Immunol., 2008-04-01;180(7):5109-17.
Species: Human
Sample Types: Whole Cells
Applications: Flow Cytometry
High-affinity autoantibodies specifically eliminate granulocyte-macrophage colony-stimulating factor activity in the lungs of patients with idiopathic pulmonary alveolar proteinosis.
Authors: Uchida K, Nakata K, Trapnell BC, Terakawa T, Hamano E, Mikami A, Matsushita I, Seymour JF, Oh-Eda M, Ishige I, Eishi Y, Kitamura T, Yamada Y, Hanaoka K, Keicho N
Blood, 2003-09-25;103(3):1089-98.
Species: Human
Sample Types: Whole Tissue
Applications: IHC-P

FAQs

Do you have a protocol for the isolation and treatment of dendritic cells used for IC flow staining with Human GM-CSF Antibody, catalog #s MAB615 and IC615?
- MagCellect Human CD14+ Cell Isolation Kit, catalog # MAGH105, is used to isolate CD14+ monocytes from PBMCs, as the starting population. Monocytes are cultured in media with rhIL-4 (catalog # 204-IL) at 20 ng/ml and rhGM-CSF (catalog # 215-GM) at 50ng/ml. On day 6, add LPS at 20ng/ml, rhTNF-a (catalog # 210-TA) at 20ng/ml and rhIL-1b (catalog # 201-LB) at 20ng/ml to culture medium and incubate cells overnight. Collect cells for flow cytometric analysis on day 7.

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