IL-8 Antibody, anti-human, REAfinity™

Name: IL-8 Antibody, anti-human, REAfinity™
Availability: InStock

IL-8 Antibody, anti-human, REAfinity™Extended ValidationRecombinant

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

Intracellular flow cytometry applications forIL-8 Antibody, anti-human, REAfinity™

APC
FITC
PE
Vio B515

Conjugate:

APC

Recommended dilution:

1:50

Remark:

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

Tested:

QC tested

Products used:

130-112-016, 130-111-864

Protocols:

Intracellular flow cytometry staining protocol (Inside Stain Kit 1:50)

Description:

Human peripheral blood mononuclear cells (PBMCs) were either left unstimulated (left peak) or stimulated with LPS for 6 hours. After two hours 1 µg/mL brefeldin A was added. Cells were then fixed, permeabilized, and stained with Anti-IL-8 antibodies as well as with CD14 antibodies. CD14⁺ cells were pre-gated for the analysis. Flow cytometry was performed using the MACSQuant^® Analyzer. Cell debris were excluded from the analysis based on scatter signals.

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 clones	Overlap 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.

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.

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

Clone	REA731
Clonality	monoclonal
Isotype	recombinant human IgG1
Isotype control	REA Control Antibody (I), human IgG1
Host	human cell line
Type of antibody	Primary antibodies, Recombinant antibodies
Species	human
Antigen	IL-8
Alternative names of antigen	Interleukin-8, Emoctakin, GCP-1, MDNCF, MONAP, NAP-1, Protein 3-10C, CXCL8
Molecular mass of antigen [kDa]	9
Distribution of antigen	T cells, basophils, neutrophils
Entrez Gene ID	3576
RRID	AB_2652456, AB_2652457, AB_2652458, AB_2652459, AB_2652460, AB_2652455

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

Certificates

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

Publications

Mukaida, N. et al. (1989) Genomic structure of the human monocyte-derived neutrophil chemotactic factor IL-8. J Immunol 143(4): 1366-1371
Matsushima, K. et al. (1989) Interleukin 8 and MCAF: novel inflammatory cytokines inducible by IL 1 and TNF. Cytokine 1(1): 2-13
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

Applications

Extended validation