Flow cytometric analysis of Islet-1 in mouse pancreatic tumor (insulinoma) cells. Beta-TC-6 (ATCC CRL-11506™) cells were fixed with BD Cytofix™ fixation buffer (Cat. No. 554655) and permeabilized with BD Phosflow™ Perm buffer III (Cat. No. 558050). The cells were stained with either PE Mouse IgG1, κ isotype control (dashed line, Cat. No. 554680) or PE Mouse anti-Islet-1 monoclonal antibody (solid line) at matched concentrations. This antibody cross-reacts with Islet-2 as well. Flow cytometry was performed on a BD FACSCanto™ II flow cytometry system.
Flow cytometric analysis of Islet-1 in cardiomyocytes derived from human embryonic stem (ES) cells. H7 human ES cells (WiCell, Madison, WI) were differentiated towards a cardiac lineage (Xu C. et al, 2011), fixed with BD Cytofix™Fixation Buffer (Cat. No. 554655) and permeabilized with BD Phosflow Perm Buffer III (Cat. No.558050). The cells were stained with either PE Mouse IgG1, κ isotype control (dashed lines, Cat. No.554680) or PE Mouse Anti-Islet-1 antibody (solid line) at matched concentrations. The histograms were derived from gated events based on light scattering characteristics of H7-derived cardiomyocytes. Flow cytometry was performed on a BD LSR™ II flow cytometry system.
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Flow cytometric analysis of Islet-1 in mouse pancreatic tumor (insulinoma) cells. Beta-TC-6 (ATCC CRL-11506™) cells were fixed with BD Cytofix™ fixation buffer (Cat. No. 554655) and permeabilized with BD Phosflow™ Perm buffer III (Cat. No. 558050). The cells were stained with either PE Mouse IgG1, κ isotype control (dashed line, Cat. No. 554680) or PE Mouse anti-Islet-1 monoclonal antibody (solid line) at matched concentrations. This antibody cross-reacts with Islet-2 as well. Flow cytometry was performed on a BD FACSCanto™ II flow cytometry system.
Flow cytometric analysis of Islet-1 in cardiomyocytes derived from human embryonic stem (ES) cells. H7 human ES cells (WiCell, Madison, WI) were differentiated towards a cardiac lineage (Xu C. et al, 2011), fixed with BD Cytofix™Fixation Buffer (Cat. No. 554655) and permeabilized with BD Phosflow Perm Buffer III (Cat. No.558050). The cells were stained with either PE Mouse IgG1, κ isotype control (dashed lines, Cat. No.554680) or PE Mouse Anti-Islet-1 antibody (solid line) at matched concentrations. The histograms were derived from gated events based on light scattering characteristics of H7-derived cardiomyocytes. Flow cytometry was performed on a BD LSR™ II flow cytometry system.
Product Details
BD Pharmingen™
ISL-1, ISL1, Islet-2, ISL-2, ISL2
Mouse (QC Testing), Human (Tested in Development)
Mouse IgG1, κ
Human Islet-1 Recombinant Protein
Intracellular staining (flow cytometry) (Routinely Tested)
5 µl
AB_11154592
Aqueous buffered solution containing BSA and ≤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 R-PE under optimum conditions, and unconjugated antibody and free PE were removed.
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.
- Source of all serum proteins is from USDA inspected abattoirs located in the United States.
- 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.
- Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
Companion Products
Stain Buffer (FBS) RUO
Size
500 mL
Cat No.
554656
Fixation Buffer RUO
Size
100 mL
Cat No.
554655
Perm Buffer III RUO
Size
125 mL
Cat No.
558050
PE Mouse IgG1, κ Isotype Control RUO
Size
0.1 mg
Cat No.
554680
562547 Rev. 2
Antibody Details
Q11-465
Islet-1 is a LIM-homeodomain transcription factor important for motor neuron differentiation and the formation of islet cells in the pancreas. Various heart cell types, such as cardiac muscle, the conduction system and endothelial cells in multiple heart tissue compartments during cardiogenesis, have been found to originate from Islet-1-positive cardiac precursor cells. Moreover, Islet-1-positive cells from differentiated human embryonic stem cell lines were found to be capable of self-renewal and expansion and could differentiate into the three major cell types of the heart.
Western blot analyses of lysates from transfected cells demonstrate that the Q11-465 monoclonal antibody reacts with human Islet-1 (ISL-1, ISL1) and Islet-2 (ISL-2, ISL2), similar to the 4D5 clone (Tsuchida et al, 1994). Cross-reactivity with mouse Islet is also observed.
562547 Rev. 2
Format Details
PE
R-Phycoerythrin (PE), is part of the BD family of Phycobiliprotein dyes. This fluorochrome is a multimeric fluorescent phycobiliprotein with excitation maximum (Ex Max) of 496 nm and 566 nm and an emission maximum (Em Max) at 576 nm. PE is designed to be excited by the Blue (488 nm), Green (532 nm) and Yellow-Green (561 nm) lasers and detected using an optical filter centered near 575 nm (e.g., a 575/26-nm bandpass filter). As PE is excited by multiple lasers, this can result in cross-laser excitation and fluorescence spillover on instruments with various combinations of Blue, Green, and Yellow-Green lasers. Please ensure that your instrument’s configurations (lasers and optical filters) are appropriate for this dye.
PE
Yellow-Green 488 nm, 532 nm, 561 nm
496 nm, 566 nm
576 nm
562547 Rev.2
Citations & References
Development References (6)
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Bu L, Jiang X, Martin-Puig S, et al. Human ISL1 heart progenitors generate diverse multipotent cardiovascular cell lineages. Nature. 2009; 460(7251):113-117. (Biology). View Reference
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Ebert AD, Yu J, Rose FF Jr, et al. Induced pluripotent stem cells from a spinal muscular atrophy patient. Nature. 2009; 457(7227):277-280. (Biology). View Reference
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Laugwitz KL, Moretti A, Caron L, Nakano A, Chien KR. Islet1 cardiovascular progenitors: a single source for heart lineages. Development. 2008; 135(2):193-205. (Biology). View Reference
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Osumi N, Hirota A, Ohuchi H, et al. Pax-6 is involved in the specification of hindbrain motor neuron subtype. Development. 1997; 124(15):2961-2972. (Biology). View Reference
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Tsuchida T, Ensini M, Morton SB, et al. Topographic organization of embryonic motor neurons defined by expression of LIM homeobox genes. Cell. 1994; 79(6):957-970. (Biology). View Reference
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Xu C, Police S, Hassanipour M, et al. Efficient generation and cryopreservation of cardiomyocytes derived from human embryonic stem cells. 2011; 6(1):53-66. (Methodology: Cell differentiation). View Reference
562547 Rev. 2
Please refer to Support Documents for Quality Certificates
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.
