anti-Human IL-6 Receptor antibody for Flow Cytometry

Recommended IL-6 Receptor Antibody (supplied by: Log in to see )

Antigen
Interleukin-6 Receptor (IL6R) Antibodies
  • IL6R
  • il-6ra
  • CD126
  • IL-6R-1
  • IL-6RA
  • IL6Q
  • IL6RA
  • IL6RQ
  • gp80
  • IL6R1
  • Il6ra
  • IL-6Ralpha
  • IL-6R
  • Il6r
  • interleukin 6 receptor
  • interleukin 6 receptor-like
  • interleukin 6 receptor, alpha
  • IL6R
  • il6r
  • Il6r
  • LOC100347752
  • Il6ra
Reactivity
Human
199
88
45
1
Host
Mouse
145
95
45
21
2
Clonality
Monoclonal
Conjugate
This IL-6 Receptor antibody is conjugated to PE-Cy7
26
17
15
12
10
9
7
7
5
5
5
5
4
4
3
3
3
3
3
2
2
2
1
1
1
1
1
1
1
1
1
1
1
Application
Flow Cytometry (FACS)
135
135
102
71
40
24
24
22
19
13
11
10
10
8
8
8
4
2
2
2
2
2
1
Supplier
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Available images

Catalog No. ABIN2659082
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Relevance Score ABIN Application Conjugate Host Isotype Epitope Supplier Clonality References Details
13.996978 ABIN4899502 CyTOF ELISA (Capture) FACS Neut WB Mouse IgG1 AA 20-339 Log in to see 17506 15
13.996978 ABIN4899503 CyTOF ELISA (Capture) FACS Neut WB Mouse IgG1 AA 20-339 Log in to see 17506 15
13.996978 ABIN4896217 FACS Allophycocyanin Mouse IgG1 AA 20-339 Log in to see 17506 1
13.996978 ABIN4896216 FACS Allophycocyanin Mouse IgG1 AA 20-339 Log in to see 17506 1
13.996978 ABIN1105828 EIA FACS Mouse IgG1 Log in to see B-N12 4
13.996978 ABIN1105829 Func IHC (fro) FACS IP Mouse IgG1 Log in to see B-R6 3
13.996978 ABIN1105827 Func EIA IHC (fro) FACS Mouse IgG1 Log in to see B-F19 2
13.996978 ABIN1383677 FACS FITC Mouse IgG1 Log in to see B-N12 1
13.996978 ABIN1105832 FACS FITC Mouse IgG1 Log in to see B-R6
13.996978 ABIN4896220 FACS PE Mouse IgG1 AA 20-339 Log in to see 17506
13.996978 ABIN4896218 FACS Fluorescein Mouse IgG1 AA 20-339 Log in to see 17506
13.996978 ABIN4896219 FACS Fluorescein Mouse IgG1 AA 20-339 Log in to see 17506
13.996978 ABIN4896221 FACS PE Mouse IgG1 AA 20-339 Log in to see 17506
13.996978 ABIN255790 ICC FACS IHC (fro) IF IP IHC ELISA Mouse IgG1 Log in to see B-R6
13.996978 ABIN2662432 FACS IF PE Mouse IgG1 kappa Log in to see UV4 5
13.996978 ABIN2659946 FACS IF IHC PerCP-Cy5.5 Mouse IgG1 kappa Log in to see UV4 3
13.996978 ABIN2659081 FACS PE-Cy7 Mouse IgG1 kappa Log in to see UV4 3
13.996978 ABIN2657623 CyTOF FACS Alexa Fluor 647 Mouse IgG1 Log in to see BL-126 3
13.996978 ABIN517051 FACS WB Mouse AA 1-468, full length Log in to see Polyclonal
13.996978 ABIN517052 FACS WB Mouse AA 1-468, full length Log in to see Polyclonal

General

Antigen Interleukin-6 Receptor (IL6R) Antibodies
Reactivity Human
(199), (88), (45), (1)
Host Mouse
(145), (95), (45), (21), (2)
Clonality (Clone)
Monoclonal   ( )
Conjugate This IL-6 Receptor antibody is conjugated to PE-Cy7
(26), (17), (15), (12), (10), (9), (7), (7), (5), (5), (5), (5), (4), (4), (3), (3), (3), (3), (3), (2), (2), (2), (1), (1), (1), (1), (1), (1), (1), (1), (1), (1), (1)
Application Flow Cytometry (FACS)
(135), (135), (102), (71), (40), (24), (24), (22), (19), (13), (11), (10), (10), (8), (8), (8), (4), (2), (2), (2), (2), (2), (1)
Pubmed 3 references available
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Product Details anti-IL-6 Receptor Antibody

Target Details IL-6 Receptor Application Details Handling References for anti-IL-6 Receptor antibody (ABIN2659082) Images
Purification The antibody was purified by affinity chromatography and conjugated with PE/Cy7 under optimal conditions. The solution is free of unconjugated PE/Cy7 and unconjugated antibody.
Clone UV4
Isotype IgG1 kappa

Target Details IL-6 Receptor

Product Details anti-IL-6 Receptor Antibody Application Details Handling References for anti-IL-6 Receptor antibody (ABIN2659082) Images back to top
Antigen
Alternative Name CD126 (IL6R Antibody Abstract)
Background CD126 is an 80 kD IL-6 receptor α chain also known as IL-6R. It is a member of the immunoglobulin superfamily that is expressed on plasma cells, T cells, activated B cells, monocytes, granulocytes, hepatocytes, epithelial cells, and fibroblasts. Functional IL-6 receptors are formed by the non-covalent association of CD126 and the IL-6 receptor β chain (CD130 or gp130). CD126 binds IL-6 with low affinity but does not signal. The β chain (gp130, CD130) does not bind IL-6 by itself but associates with the α-chain/IL-6 complex to initiate signal transduction. IL-6 binding to the receptor complex results in the stimulation of B and T cells, and hematopoietic precursor proliferation and differentiation. A soluble form of CD126 has been found in human serum.
Pathways JAK-STAT Signaling, Autophagy, Growth Factor Binding

Application Details

Product Details anti-IL-6 Receptor Antibody Target Details IL-6 Receptor Handling References for anti-IL-6 Receptor antibody (ABIN2659082) Images back to top
Application Notes Optimal working dilution should be determined by the investigator.
Restrictions For Research Use only

Handling

Product Details anti-IL-6 Receptor Antibody Target Details IL-6 Receptor Application Details References for anti-IL-6 Receptor antibody (ABIN2659082) Images back to top
Buffer Phosphate-buffered solution, pH 7.2, containing 0.09 % sodium azide and 0.2 % (w/v) BSA .
Preservative Sodium azide
Precaution of Use This product contains Sodium azide: a POISONOUS AND HAZARDOUS SUBSTANCE which should be handled by trained staff only.
Handling Advice Protect from prolonged exposure to light. Do not freeze.
Storage 4 °C
Storage Comment The antibody solution should be stored undiluted between 2°C and 8°C.

References for anti-IL-6 Receptor antibody (ABIN2659082)

Product Details anti-IL-6 Receptor Antibody Target Details IL-6 Receptor Application Details Handling Images back to top
Background publications

Boulanger, Chow, Brevnova, Garcia: "Hexameric structure and assembly of the interleukin-6/IL-6 alpha-receptor/gp130 complex." in: Science (New York, N.Y.), Vol. 300, Issue 5628, pp. 2101-4, 2003

Taga, Kishimoto: "Gp130 and the interleukin-6 family of cytokines." in: Annual review of immunology, Vol. 15, pp. 797-819, 1997

Gaillard, Bataille, Brailly, Zuber, Yasukawa, Attal, Maruo, Taga, Kishimoto, Klein: "Increased and highly stable levels of functional soluble interleukin-6 receptor in sera of patients with monoclonal gammopathy." in: European journal of immunology, Vol. 23, Issue 4, pp. 820-4, 1993

'Independent Validation' Badge
Antigen IL6R
Lot Number B236171
Method validated Flow Cytometry
Positive Control Mature dendritic cells (mDCs) from three donors, incubated with ABIN2659082
Negative Control Mature dendritic cells (mDCs) from three donors, incubated without antibody
Notes

Passed. ABIN2659082 specifically recognizes the IL-6 Receptor on mDCs.

Primary Antibody ABIN2659082
Protocol
  • Prepare peripheral blood mononuclear cells (PBMCs) from leukoreduction system chambers (LRSCs) (Pfeiffer et al. (2013); Weidinger et al. 2011) from three healthy donors (obtained following informed consent and approved by the institutional review board) by density centrifugation on a Lymphoprep gradient (Axis-shield AS, 1114547, lot 12IFS10).
  • For the generation of dendritic cells (DCs), prepare PBMCs from leukoreduction system chambers (LRSC) (Pfeiffer et al. (2013)).
  • After centrifugation, collect PBMCs forming a distinct band at the sample/Lymphoprep interface.
  • Wash PBMCs 3x with cold PBS supplemented with 1mM EDTA.
  • Wash PBMCs 1x with cold RPMI 1640 (Lonza, BE12-167F, lot 7MB088).
  • Resuspend PBMCs in warm DC Medium (RPMI 1640 supplemented with 1% AB-serum, 100U/ml penicillin, 100mg/ml streptomycin, 2mM L-glutamine, 10mM HEPES buffer (Lonza, BE17-737E)) and seeded into 175cm2 tissue culture flasks at a density of 3.5x108 cells/flask.
  • Let monocytes adhere for 1h at 37°C and 5% CO2.
  • Wash non-adherent fraction off with warm RPMI 1640.
  • Add 30ml DC medium supplemented with 800U/ml recombinant human granulocyte-macrophage colony-stimulating factor (GM-CSF) and 250U/ml recombinant human interleukin-4 (IL-4) to each flask (day 1).
  • Incubate cells for 72h at 37°C and 5% CO2.
  • On day 4 (or three days later), add 5ml of fresh culture medium containing GM-CSF (final concentration of 400 U/ml in 35ml) and IL-4 (final concentration of 250U/ml in 35ml) to the cells.
  • On day 5, mature the immature DCs by adding 10ng/ml recombinant human tumor necrosis factor alpha (TNF-α), 1μg/ml prostaglandin E2 (PGE2), 200U/ml recombinant human interleukin-1β (IL-1β), 1000U/ml recombinant human interleukin-6 (IL-6), 40U/ml GM-CSF, and 250 U/ml IL-4 to the medium.
  • Incubate cells for 48h at 37°C and 5% CO2.
  • On day 7, use the now mature DCs (mDCs) for FACS analysis.
  • Analyze cell surface expression of IL-6 Receptor by FACS:
    • Harvest 2.5x105 mDCs by centrifugation.
    • Wash cells once with FACS buffer (PBS supplemented with 2% FCS).
    • Combine anti-human PE-Cy7 conjugated IL-6 Receptor antibody (antibodies-online, ABIN2659082, lot B236171) diluted 1:100 and LIVE/DEAD Violet dead cell stain kit (ThermoFisher Scientific, L34955, lot 1832693) diluted 1:300 in cold FACS buffer.
    • Resuspend mDCs in 100µl of the antibody-solution and incubate for 30min on ice in the dark.
    • Wash mDCs 2x with cold FACS buffer.
    • Fix mDCs using 2% paraformaldehyde in cold PBS.
    • Store mDCs at 4°C until analysis.
    • Run samples using a BD FACS Canto II and analyze events using FCS Express 5.
Experimental Notes

Mature DCs were stained for FACS for the analysis of the expression of IL-6 Receptor/CD126 on the cell surface of the cells with ABIN2659082 for 30min or 120min respectively. The histogram shows a clearly shift between unstained control and the stained mock control by staining for 120min. The staining for 30min does also show a shift of the histogram peak, but not as clear as the staining for 120min.

Validation Images
Flow Cytometry validation image for anti-Interleukin-6 Receptor (IL6R) antibody (PE-Cy7) (ABIN2659082)

A Flow cytometry analysis of mature DCs from three different donors after incubati...