Flow cytometric analysis of MAP2B expression on Neurons. H9 human embryonic stem (ES) cells (WiCell, Madison, WI) were differentiated into Neural Precursor cells (NPCs) and grown for 4 passages before differentiating into neurons and glia for 12 days. The cells were fixed with Fixation buffer (Cat. No. 554655), permeabilized with Perm Buffer III (Cat. No. 558050), and then stained with either Alexa Fluor® 488 Mouse anti-MAP2B (Cat. No. 560399, solid line) or Alexa Fluor® 488 Mouse IgG1,k Isotype Control (Cat. No. 557721, dashed line). This antibody also works in Perm/Wash Buffer I (Cat. No. 557885). The fluorescence histograms were derived from gated events with the forward and side light-scattering characteristics of viable neurons. Flow cytometry was performed on a BD LSR™ II.
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Flow cytometric analysis of MAP2B expression on Neurons. H9 human embryonic stem (ES) cells (WiCell, Madison, WI) were differentiated into Neural Precursor cells (NPCs) and grown for 4 passages before differentiating into neurons and glia for 12 days. The cells were fixed with Fixation buffer (Cat. No. 554655), permeabilized with Perm Buffer III (Cat. No. 558050), and then stained with either Alexa Fluor® 488 Mouse anti-MAP2B (Cat. No. 560399, solid line) or Alexa Fluor® 488 Mouse IgG1,k Isotype Control (Cat. No. 557721, dashed line). This antibody also works in Perm/Wash Buffer I (Cat. No. 557885). The fluorescence histograms were derived from gated events with the forward and side light-scattering characteristics of viable neurons. Flow cytometry was performed on a BD LSR™ II.
Product Details
BD Pharmingen™
Human (QC Testing), Rat (Tested in Development), Mouse (Reported)
Mouse IgG1, κ
Human MAP2B aa. 19-219 Recombinant Protein
Intracellular staining (flow cytometry) (Routinely Tested)
20 µl
AB_1645358
Aqueous buffered solution containing BSA, protein stabilizer, 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 to Alexa Fluor® 488 under optimum conditions, and unreacted Alexa Fluor® 488 was 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.
- Alexa Fluor® 488 fluorochrome emission is collected at the same instrument settings as for fluorescein isothiocyanate (FITC).
- The Alexa Fluor®, Pacific Blue™, and Cascade Blue® dye antibody conjugates in this product are sold under license from Molecular Probes, Inc. for research use only, excluding use in combination with microarrays, or as analyte specific reagents. The Alexa Fluor® dyes (except for Alexa Fluor® 430), Pacific Blue™ dye, and Cascade Blue® dye are covered by pending and issued patents.
- Alexa Fluor® is a registered trademark of Molecular Probes, Inc., Eugene, OR.
- Species cross-reactivity detected in product development may not have been confirmed on every format and/or application.
- Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
Companion Products
Stain Buffer (FBS) RUO
Size
500 mL
Cat No.
554656
Stain Buffer (BSA) RUO
Size
500 mL
Cat No.
554657
Perm/Wash Buffer I RUO
Size
125 mL
Cat No.
557885
Fixation Buffer RUO
Size
100 mL
Cat No.
554655
Perm Buffer III RUO
Size
125 mL
Cat No.
558050
Alexa Fluor® 488 Mouse IgG1 κ Isotype Control RUO
Size
100 Tests
Cat No.
557721
560399 Rev. 2
Antibody Details
18/MAP2B
Microtubule-associated proteins (MAPs) play a crucial role in the development and morphology of neurons. MAP2, specifically localized in dendrites, has four known isoforms that are produced by alternative splicing of the transcript and are expressed at various stages of neuronal development. MAP2B is a 280-kDa protein that is expressed throughout brain development. It contains functional domains that interact with the regulatory subunit of the cAMP-dependent kinase II and microtubules. Experimental monitoring of its presence, along with GFAP and nestin, may be utilized to quantify neuronal development.
560399 Rev. 2
Format Details
Alexa Fluor™ 488
Alexa Fluor™ 488 Dye is part of the BD blue family of dyes. This is a small organic fluorochrome with an excitation maximum (Ex Max) at 494-nm and an emission maximum (Em Max) at 517-nm. Alexa Fluor™ 488 is designed to be excited by the Blue laser (488 nm) and detected using an optical filter centered near 520-nm (e.g., a 530/30-nm bandpass filter). Please ensure that your instrument’s configurations (lasers and optical filters) are appropriate for this dye.
Alexa Fluor™ 488
Blue 488 nm
494 nm
517 nm
560399 Rev.2
Citations & References
Development References (3)
-
Charlesworth P, Komiyama NH, Grant SGN. Homozygous mutation of focal adhesion kinase in embryonic stem cell derived neurons: normal electrophysiological and morphological properties in vitro. BMC Neurosci. 2006; 7:47. (Clone-specific: Immunofluorescence). View Reference
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Kanaani J, el-Husseini Ael-D, Aguilera-Moreno A, Diacovo JM, Bredt DS, Baekkeskov S. A combination of three distinct trafficking signals mediates axonal targeting and presynaptic clustering of GAD65. J Cell Biol. 2002; 158(7):1229-1238. (Clone-specific). View Reference
-
Kindler S, Schulz B, Goedert M, Garner CC. Molecular structure of microtubule-associated protein 2b and 2c from rat brain. J Biol Chem. 1990; 265(32):19679-19684. (Biology: Immunofluorescence). View Reference
560399 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.
