GM-CSF Antibody, anti-human

Name: GM-CSF Antibody, anti-human
Availability: InStock

GM-CSF Antibody, anti-humanExtended Validation

Clone:

BVD2-21C11

Type of antibody:

Primary antibodies, Functional-grade antibodies

Isotype:

rat IgG2aκ

Applications:

FA, ICFC

Alternative names:

CSF2, GM-CSF

Intracellular flow cytometry applications forGM-CSF Antibody, anti-human

APC
PE

Conjugate:

APC

Recommended dilution:

1:10

Remark:

The antibody is suited for staining of formaldehyde-fixed cells.

Tested:

QC tested

Products used:

130-099-625, 130-096-078

Protocols:

Intracellular flow cytometry staining protocol (Inside Stain Kit 1:10, protocol A)

Description:

Human peripheral blood mononuclear cells (PBMCs) were either left unstimulated (left images) or stimulated with PMA/ionomycin for 6 hours. After 2 hours, brefeldin A was added. Cells were then fixed, permeabilized, and intracellularly stained with Anti-GMCSF antibodies and CD4 antibodies and analyzed by flow cytometry using the MACSQuant^® Analyzer. Gating was performed according to forward and side scatter properties of the cells.

Functional assay applications forGM-CSF Antibody, anti-human

pure-functional grade

Conjugate:

pure-functional grade

Tested:

during development

Products used:

130-096-075

Type of functional assay:

neutralization of cytokine activity

Extended validation for GM-CSF Antibody, anti-human

Specificity

Epitope competition

In order to compare the epitope specificity of an antibody, the clone being used is compared with other known clones recognizing the same antigen in a competition assay.

Other clones	Overlap in epitope recognition with BVD2-21C11
REA1215	++
DAVKAT	-
Cells were incubated with an excess of purified unconjugated GM-CSF (BVD2-21C11) antibody followed by staining with fluorochrome-conjugated antibodies of other known clones against the same marker. Based on the fluorescence signal obtained, the clones were identified as recognizing completely overlapping (++), partially overlapping (+), or completely different epitopes (-) of the marker.

Sensitivity

Performance comparison

Selected fluorochrome conjugated antibodies from Miltenyi Biotec were compared to commercially available hybridoma clones in flow cytometry analysis.

Flow cytometric comparison of different clones for GM-CSF. Human peripheral blood mononuclear cells (PBMCs) were stimulated with PMA & Ionomycin and were stained with GM-CSF antibodies and with a suitable counterstaining. Cells were then fixed and permeabilized using the Inside Stain Kit followed by a staining with (specificity from PIM) antibodies. As a control, GM-CSF antibody staining was omitted and cells were measured in the same channels. Flow cytometry was performed with the MACSQuant^® Analyzer. Cell debris, dead cells, and cell doublets were excluded from the analysis based on scatter signals and Viobility 405/520 Fixable Dye fluorescence. No FcR Blocking Reagent was used. The recommended titers of respective antibodies from different suppliers were used.

Flow cytometric comparison of different clones for GM-CSF. Human peripheral blood mononuclear cells (PBMCs) were stimulated with PMA & Ionomycin and were stained with GM-CSF antibodies and with a suitable counterstaining. Cells were then fixed and permeabilized using the Inside Stain Kit followed by a staining with (specificity from PIM) antibodies. As a control, GM-CSF antibody staining was omitted and cells were measured in the same channels. Flow cytometry was performed with the MACSQuant^® Analyzer. Cell debris, dead cells, and cell doublets were excluded from the analysis based on scatter signals and Viobility 405/520 Fixable Dye fluorescence. No FcR Blocking Reagent was used. The recommended titers of respective antibodies from different suppliers were used.

Flow cytometric comparison of different clones for GM-CSF. Human peripheral blood mononuclear cells (PBMCs) were stimulated with PMA & Ionomycin and were stained with GM-CSF antibodies and with a suitable counterstaining. Cells were then fixed and permeabilized using the Inside Stain Kit followed by a staining with (specificity from PIM) antibodies. As a control, GM-CSF antibody staining was omitted and cells were measured in the same channels. Flow cytometry was performed with the MACSQuant^® Analyzer. Cell debris, dead cells, and cell doublets were excluded from the analysis based on scatter signals and Viobility 405/520 Fixable Dye fluorescence. No FcR Blocking Reagent was used. The recommended titers of respective antibodies from different suppliers were used.

Flow cytometric comparison of different clones for GM-CSF. Human peripheral blood mononuclear cells (PBMCs) were stimulated with PMA & Ionomycin and were stained with GM-CSF antibodies and with a suitable counterstaining. Cells were then fixed and permeabilized using the Inside Stain Kit followed by a staining with (specificity from PIM) antibodies. As a control, GM-CSF antibody staining was omitted and cells were measured in the same channels. Flow cytometry was performed with the MACSQuant^® Analyzer. Cell debris, dead cells, and cell doublets were excluded from the analysis based on scatter signals and Viobility 405/520 Fixable Dye fluorescence. No FcR Blocking Reagent was used. The recommended titers of respective antibodies from different suppliers were used.

Flow cytometric comparison of different clones for GM-CSF. Human peripheral blood mononuclear cells (PBMCs) were stimulated with PMA & Ionomycin and were stained with GM-CSF antibodies and with a suitable counterstaining. Cells were then fixed and permeabilized using the Inside Stain Kit followed by a staining with (specificity from PIM) antibodies. As a control, GM-CSF antibody staining was omitted and cells were measured in the same channels. Flow cytometry was performed with the MACSQuant^® Analyzer. Cell debris, dead cells, and cell doublets were excluded from the analysis based on scatter signals and Viobility 405/520 Fixable Dye fluorescence. No FcR Blocking Reagent was used. The recommended titers of respective antibodies from different suppliers were used.

Specifications for GM-CSF Antibody, anti-human

Overview

Granulocyte macrophage colony-stimulating factor (GM‑CSF) is a hematopoietic growth factor that is essential for proliferation and development of granulocyte and monocyte/macrophage progenitors. It also functions as a growth factor for erythroid and megakaryocytic precursor cells in conjunction with erythropoietin. GM-CSF is secreted by various cell types including T cells, macrophages, endothelial cells, and fibroblasts in response to inflammatory stimuli and cytokines. In addition, GM-CSF strongly chemoattracts neutrophils and eosinophils and enhances the effector functions of neutrophils and macrophages.

Alternative names

CSF2, GM-CSF

Detailed product information

Technical specifications

Clone	BVD2-21C11
Clonality	monoclonal
Isotype	rat IgG2aκ
Isotype control	Isotype Control Antibody, rat IgG2a
Host	rat
Type of antibody	Primary antibodies, Functional-grade antibodies
Species	human, non-human primate
Cross-reactivity	rhesus monkey ( Macaca mulatta ) , cynomolgus monkey ( Macaca fascicularis )
Antigen	GM-CSF
Alternative names of antigen	CSF2, GM-CSF
Molecular mass of antigen [kDa]	14
Entrez Gene ID	1437
RRID	AB_2651852, AB_2651853, AB_2651849, AB_2651851

Resources for GM-CSF Antibody, anti-human

Certificates

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Applications

Extended validation