Clone:
REA731
Type of antibody:
Primary antibodies, Recombinant antibodies
Isotype:
recombinant human IgG1
Applications:
ICFC
Alternative names:
Interleukin-8, Emoctakin, GCP-1, MDNCF, MONAP, NAP-1, Protein 3-10C, CXCL8

Extended validation for IL-8 Antibody, anti-human, REAfinity™

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 clonesOverlap in epitope recognition with REA731
AS14++
G265-8++
E8N1-
BH0814-
6217-
Cells were incubated with an excess of purified unconjugated IL-8 (REA731) 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.
View details
Flow cytometric comparison of different clones for IL-8. Human peripheral blood mononuclear cells (PBMCs) were stimulated with 10ng/ml LPS for 6h and afterwards treated with 1µg/ml Brefaldin A for 2h.Cells were first stained with Viobility™ Fixable Dye followed by a suitable counterstaining. Cells were then fixed and permeabilized using the Inside Stain Kit followed by a staining with IL-8 antibodies. As a control, IL-8antibody staining was omitted and cells were measured in the same channels. Flow cytometry was performed with the MACSQuant® Analyzer. Cells were pregated on CD14+ cells. 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.
View details
Flow cytometric comparison of different clones for IL-8. Human peripheral blood mononuclear cells (PBMCs) were stimulated with 10ng/ml LPS for 6h and afterwards treated with 1µg/ml Brefaldin A for 2h.Cells were first stained with Viobility™ Fixable Dye followed by a suitable counterstaining. Cells were then fixed and permeabilized using the Inside Stain Kit followed by a staining with IL-8 antibodies. As a control, IL-8antibody staining was omitted and cells were measured in the same channels. Flow cytometry was performed with the MACSQuant® Analyzer. Cells were pregated on CD14+ cells. 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.
View details
Flow cytometric comparison of different clones for IL-8. Human peripheral blood mononuclear cells (PBMCs) were stimulated with 10ng/ml LPS for 6h and afterwards treated with 1µg/ml Brefaldin A for 2h.Cells were first stained with Viobility™ Fixable Dye followed by a suitable counterstaining. Cells were then fixed and permeabilized using the Inside Stain Kit followed by a staining with IL-8 antibodies. As a control, IL-8antibody staining was omitted and cells were measured in the same channels. Flow cytometry was performed with the MACSQuant® Analyzer. Cells were pregated on CD14+ cells. 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.
View details
Flow cytometric comparison of different clones for IL-8. Human peripheral blood mononuclear cells (PBMCs) were stimulated with 10ng/ml LPS for 6h and afterwards treated with 1µg/ml Brefaldin A for 2h.Cells were first stained with Viobility™ Fixable Dye followed by a suitable counterstaining. Cells were then fixed and permeabilized using the Inside Stain Kit followed by a staining with IL-8 antibodies. As a control, IL-8antibody staining was omitted and cells were measured in the same channels. Flow cytometry was performed with the MACSQuant® Analyzer. Cells were pregated on CD14+ cells. 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 IL-8. Human peripheral blood mononuclear cells (PBMCs) were stimulated with 10ng/ml LPS for 6h and afterwards treated with 1µg/ml Brefaldin A for 2h.Cells were first stained with Viobility™ Fixable Dye followed by a suitable counterstaining. Cells were then fixed and permeabilized using the Inside Stain Kit followed by a staining with IL-8 antibodies. As a control, IL-8antibody staining was omitted and cells were measured in the same channels. Flow cytometry was performed with the MACSQuant® Analyzer. Cells were pregated on CD14+ cells. 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 IL-8 Antibody, anti-human, REAfinity™

Overview

Clone REA731 recognizes the human IL-8 chemokine, a heterodimer also known as interleukin 8 or CXCL8 (chemokine (C-X-C motif) ligand 8. IL-8 is secreted by macrophages, epithelial cells, airway smooth muscle cells, and endothelial cells. IL-8 plays a role in neutrophil activation, is a potent promoter of angiogenesis, and attracts neutrophils, basophils, and T cells. Clone REA731 recognizes both the 72 and 77 amino acid isoforms of human IL-8.
Additional information: Clone REA731 displays negligible binding to Fc receptors.

Alternative names

Interleukin-8, Emoctakin, GCP-1, MDNCF, MONAP, NAP-1, Protein 3-10C, CXCL8

Detailed product information

Technical specifications

CloneREA731
Clonalitymonoclonal
Isotyperecombinant human IgG1
Isotype controlREA Control Antibody (I), human IgG1
Hostcell line
Type of antibodyPrimary antibodies, Recombinant antibodies
Specieshuman
AntigenIL-8
Alternative names of antigenInterleukin-8, Emoctakin, GCP-1, MDNCF, MONAP, NAP-1, Protein 3-10C, CXCL8
Molecular mass of antigen [kDa]9
Distribution of antigenT cells, basophils, neutrophils
Entrez Gene ID3576
RRIDAB_2652456, AB_2652457, AB_2652458, AB_2652459, AB_2652460, AB_2652455

References for IL-8 Antibody, anti-human, REAfinity™

Publications

  1. Mukaida, N. et al. (1989) Genomic structure of the human monocyte-derived neutrophil chemotactic factor IL-8. J. Immunol. 143(4): 1366-1371
  2. Matsushima, K. et al. (1989) Interleukin 8 and MCAF: novel inflammatory cytokines inducible by IL 1 and TNF. Cytokine 1(1): 2-13
  3. Hébert, C. A. et al. (1990) Endothelial and leukocyte forms of IL-8. Conversion by thrombin and interactions with neutrophils. J. Immunol. 145(9): 3033-3040

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