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BD Horizon™ BB515 Mouse Anti-Human CD126
Clone M5 (RUO)
Multiparameter flow cytometric analysis of CD126 expression on human peripheral blood leukocytes. Human whole blood was stained with either BB515 Mouse IgG1, κ Isotype Control (Cat. No. 564416; left panel) or BB515 Mouse Anti-Human CD126 (Cat. No. 564623; right panel). Erythrocytes were lysed with Lysing Buffer (Cat. No. 555899). Flow cytometric contour plots showing the correlated expression of CD126 (or Ig Isotype control staining) versus side light-scatter signals (SSC) were derived from gated events with the forward and side light-scatter characteristics of viable leukocyte populations. Flow cytometric analysis was performed on a BD™ LSRFortessa.
Multiparameter flow cytometric analysis of CD126 expression on human peripheral blood leukocytes. Human whole blood was stained with either BB515 Mouse IgG1, κ Isotype Control (Cat. No. 564416; left panel) or BB515 Mouse Anti-Human CD126 (Cat. No. 564623; right panel). Erythrocytes were lysed with Lysing Buffer (Cat. No. 555899). Flow cytometric contour plots showing the correlated expression of CD126 (or Ig Isotype control staining) versus side light-scatter signals (SSC) were derived from gated events with the forward and side light-scatter characteristics of viable leukocyte populations. Flow cytometric analysis was performed on a BD™ LSRFortessa.
Multiparameter flow cytometric analysis of CD126 expression on human peripheral blood leukocytes. Human whole blood was stained with either BB515 Mouse IgG1, κ Isotype Control (Cat. No. 564416; left panel) or BB515 Mouse Anti-Human CD126 (Cat. No. 564623; right panel). Erythrocytes were lysed with Lysing Buffer (Cat. No. 555899). Flow cytometric contour plots showing the correlated expression of CD126 (or Ig Isotype control staining) versus side light-scatter signals (SSC) were derived from gated events with the forward and side light-scatter characteristics of viable leukocyte populations. Flow cytometric analysis was performed on a BD™ LSRFortessa.
Multiparameter flow cytometric analysis of CD126 expression on human peripheral blood leukocytes. Human whole blood was stained with either BB515 Mouse IgG1, κ Isotype Control (Cat. No. 564416; left panel) or BB515 Mouse Anti-Human CD126 (Cat. No. 564623; right panel). Erythrocytes were lysed with Lysing Buffer (Cat. No. 555899). Flow cytometric contour plots showing the correlated expression of CD126 (or Ig Isotype control staining) versus side light-scatter signals (SSC) were derived from gated events with the forward and side light-scatter characteristics of viable leukocyte populations. Flow cytometric analysis was performed on a BD™ LSRFortessa.
Product Details
BD Horizon™
Interleukin 6 Receptor alpha chain; IL-6R alpha; IL-6Rα
Human (QC Testing)
Mouse BALB/c IgG1, κ
CD126 Recombinant Protein
Flow cytometry (Routinely Tested)
5 µl
VI C63; IX 36
AB_2738870
Aqueous buffered solution containing ≤0.09% sodium azide.
RUO
Preparation And Storage
Store undiluted at 4°C and protected from prolonged exposure to light. Do not freeze. The monoclonal antibody was purified from tissue culture supernatant or ascites by affinity chromatography. The antibody was conjugated with BD Horizon™ BB515 under optimum conditions and unconjugated antibody was removed.
Recommended Assay Procedures
BD™ CompBeads can be used as surrogates to assess fluorescence spillover (Compensation). When fluorochrome conjugated antibodies are bound to CompBeads, they have spectral properties very similar to cells. However, for some fluorochromes there can be small differences in spectral emissions compared to cells, resulting in spillover values that differ when compared to biological controls. It is strongly recommended that when using a reagent for the first time, users compare the spillover on cells and CompBead to ensure that BD Comp beads are appropriate for your specific cellular application.
For optimal and reproducible results, BD Horizon Brilliant Stain Buffer should be used anytime two or more BD Horizon Brilliant dyes are used in the same experiment. Fluorescent dye interactions may cause staining artifacts which may affect data interpretation. The BD Horizon Brilliant Stain Buffer was designed to minimize these interactions. More information can be found in the Technical Data Sheet of the BD Horizon Brilliant Stain Buffer (Cat. No. 563794/566349) or the BD Horizon Brilliant Stain Buffer Plus (Cat. No. 566385).
For optimal results, it is recommended to perform 2 washes after staining with antibodies. Cells may be prepared, stained with antibodies and washed twice with wash buffer per established protocols for immunofluorescence staining, prior to acquisition on a flow cytometer. Performing fewer than the recommended wash steps may lead to increased spread of the negative population.
Product Notices
- This reagent has been pre-diluted for use at the recommended Volume per Test. We typically use 1 × 10^6 cells in a 100-µl experimental sample (a test).
- An isotype control should be used at the same concentration as the antibody of interest.
- Caution: Sodium azide yields highly toxic hydrazoic acid under acidic conditions. Dilute azide compounds in running water before discarding to avoid accumulation of potentially explosive deposits in plumbing.
- For fluorochrome spectra and suitable instrument settings, please refer to our Multicolor Flow Cytometry web page at www.bdbiosciences.com/colors.
- BD Horizon Brilliant Stain Buffer is covered by one or more of the following US patents: 8,110,673; 8,158,444; 8,575,303; 8,354,239.
- Please refer to http://regdocs.bd.com to access safety data sheets (SDS).
- Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
Companion Products
Brilliant Stain Buffer RUO
Size
1000 Tests
Cat No.
566349
Brilliant Stain Buffer Plus RUO
Size
1000 Tests
Cat No.
566385
Stain Buffer (FBS) RUO
Size
500 ML
Cat No.
554656
Stain Buffer (BSA) RUO
Size
500 ML
Cat No.
554657
Brilliant Stain Buffer RUO
Size
100 Tests
Cat No.
563794
Lysing Solution 10X Concentrate IVD
Size
100 ML
Cat No.
349202
564623 Rev. 3
Antibody Details
M5
The M5 monoclonal antibody specifically binds to human CD126 which is also known as the alpha subunit of the human IL-6 Receptor (IL-6Rα). CD126 is an 80 kDa type I transmembrane glycoprotein, also known as gp80 and B cell stimulatory factor-2 (BSF-2) Receptor. The IL-6Rα subunit associates with the 130-160 kDa gp130 subunit (IL-6 Receptor β chain, CD130), that is shared with the receptor complexes for Leukemia Inhibitory Factor (LIF), Ciliary Neurotropic Factor (CNTF), Oncostatin M (OSM), IL-11, Cardiotropin 1 (CT-1) and possibly Neurotrophin-1/B Cell-Stimulating Factor 3 (NNT-1/BSF-3). The IL-6Rα chain binds IL-6 with low affinity; however the association with CD130 stabilizes the IL-6/IL-6Rα complex resulting in the formation of a high affinity ligand-receptor complex. The IL-6Rβ chain mediates signal transduction. CD126 is expressed at high levels by activated and EBV-transformed B cells, plasma cells and myeloma cells and at lower levels by most leucocytes, epithelial cells, fibroblasts, hepatocytes and neural cells. IL-6Rα exists in soluble form in human serum. The serum levels of soluble IL-6Rα appear to elevate in pathological situations such as multiple myeloma, Grave's disease, juvenile chronic arthritis and HIV. The M5 antibody is directed against an epitope not involved in interactions of CD126 with IL-6 or CD130.
The antibody was conjugated to BD Horizon BB515 which is part of the BD Horizon Brilliant™ Blue family of dyes. With an Ex Max near 490 nm and an Em Max near 515 nm, BD Horizon BB515 can be excited by the blue laser (488 nm) laser and detected with a 530/30 nm filter. This dye has been exclusively developed by BD Biosciences and is up to seven times brighter than FITC with less spillover into the PE channel. Due to similar excitation and emission properties, BB515, FITC, and Alexa Fluor® 488 cannot be used simultaneously. It is not recommended to use BB515 in cocktails that include Streptavidin conjugates as it may cause high background.
564623 Rev. 3
Format Details
BB515
The BD Horizon Brilliant™ Blue 515 (BB515) dye is part of the BD Horizon Brilliant™ Blue family of dyes. This dye is a polymer fluorochrome with an excitation maximum (Ex Max) at 490-nm and an emission maximum (Em Max) of 515-nm. Driven by BD innovation, BB515 is designed to be excited by the blue laser (488-nm) and detected using an optical filter centered near 520-nm (e.g., 530/30-nm). BB515 reagents are significantly brighter than equivalent FITC or Alexa Fluor™ 488 reagents with less spillover into the PE detector. Please ensure that your instrument’s configurations (lasers and optical filters) are appropriate for this dye. It is not recommended to use BB515 in cocktails that include streptavidin conjugates as it may cause high background.
BB515
490 nm
515 nm
564623 Rev.3
Citations & References
View product citations for antibody "564623" on CiteAb
Development References (7)
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Autissier P, Liautard J, Brochier J, Gaillard JP. Activation of the gp130 signaling pathway by monoclonal antibodies directed against the gp130 molecule. Eur J Immunol. 1997; 27(3):794-797. (Biology). View Reference
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Brochier J, Liautard J, Jacquet C, Gaillard JP, Klein B. Optimizing therapeutic strategies to inhibit circulating soluble target molecules with monoclonal antibodies: example of the soluble IL-6 receptors. Eur J Immunol. 2001; 31(1):259-264. (Clone-specific: ELISA). View Reference
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Cabezon R, Sintes J, Llinas L, Benitez-Ribas D. Analysis of HLDA9 mAbs on plasmacytoid dendritic cell. Immunol Lett. 2011; 134(2):167-173. (Clone-specific: Flow cytometry). View Reference
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Gaillard JP, Liautard J, Duperray C, Brochier J. mAb against human gp80 IL-6 receptor. In: Schlossman SF. Stuart F. Schlossman .. et al., ed. Leucocyte typing V : white cell differentiation antigens : proceedings of the fifth international workshop and conference held in Boston, USA, 3-7 November, 1993. Oxford: Oxford University Press; 1995:1891-1894.
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Gaillard JP, Mani JC, Liautard J, Klein B, Brochier J. Interleukin-6 receptor signaling. I. gp80 and gp130 receptor interaction in the absence of interleukin-6.. Eur Cytokine Netw. 1999; 10(1):43-8. (Clone-specific: ELISA, Functional assay). View Reference
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Llinas L, Lazaro A, de Salort J, Matesanz-Isabel J, Sintes J, Engel P. Expression profiles of novel cell surface molecules on B-cell subsets and plasma cells as analyzed by flow cytometry. Immunol Lett. 2011; 134(2):113-121. (Biology). View Reference
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Tupitsyn N, Kadagidze Z, Gaillard JP, et al. Functional interaction of the gp80 and gp130 IL-6 receptors in human B cell malignancies. Clin Lab Haematol. 1998; 20(6):345-352. (Clone-specific). View Reference
564623 Rev. 3
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Global - Refer to manufacturer's instructions for use and related User Manuals and Technical data sheets before using this products as described
Comparisons, where applicable, are made against older BD Technology, manual methods or are general performance claims. Comparisons are not made against non-BD technologies, unless otherwise noted.
For Research Use Only. Not for use in diagnostic or therapeutic procedures.