Analysis of MAP2B in H9 cells. 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 (BD Cytofix™ buffer, Cat. No. 554655) for 20 minutes at room temperature, permeabilized with BD™ Phosflow Perm Buffer I (Cat. No. 557885), and then stained with either Alexa Fluor® 647 Mouse anti-MAP2B (solid line) or Alexa Fluor® 647 Mouse IgG1 κ Isotype Control (Cat. No. 557732, dashed line). This antibody also works in BD™ PhosFlow Perm Buffer II or III (Cat. No. 558052 or 558050, respectively). Flow cytometry was performed on a BD FACSCanto™ II flow cytometry system.
Analysis of MAP2B in H9 cells. 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 (BD Cytofix™ buffer, Cat. No. 554655) for 20 minutes at room temperature, permeabilized with BD™ Phosflow Perm Buffer I (Cat. No. 557885), and then stained with either Alexa Fluor® 647 Mouse anti-MAP2B (solid line) or Alexa Fluor® 647 Mouse IgG1 κ Isotype Control (Cat. No. 557732, dashed line). This antibody also works in BD™ PhosFlow Perm Buffer II or III (Cat. No. 558052 or 558050, respectively). Flow cytometry was performed on a BD FACSCanto™ II flow cytometry system.
Product Details
BD Pharmingen™
Human (QC Testing), Mouse (Reported)
Mouse IgG1, κ
Human MAP2B aa. 19-219 Recombinant Protein
Intracellular staining (flow cytometry) (Routinely Tested)
20 µl
AB_1645423
Aqueous buffered solution containing BSA, protein stabilizer, and ≤0.09% sodium azide.
RUO
Preparation And Storage
The monoclonal antibody was purified from tissue culture supernatant or ascites by affinity chromatography. The antibody was conjugated to Alexa Fluor® 647 under optimum conditions, and unreacted Alexa Fluor® 647 was removed. Store undiluted at 4°C and protected from prolonged exposure to light. Do not freeze.
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).
- Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
- For fluorochrome spectra and suitable instrument settings, please refer to our Multicolor Flow Cytometry web page at www.bdbiosciences.com/colors.
- Alexa Fluor® 647 fluorochrome emission is collected at the same instrument settings as for allophycocyanin (APC).
- 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.
- 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.
Companion Products
Fixation Buffer RUO
Size
100 mL
Cat No.
554655
Perm/Wash Buffer I RUO
Size
125 mL
Cat No.
557885
Perm Buffer III RUO
Size
125 mL
Cat No.
558050
Perm Buffer II RUO
Size
125 mL
Cat No.
558052
Alexa Fluor® 647 Mouse IgG1 κ Isotype Control RUO
Size
100 Tests
Cat No.
557732
560382 Rev. 1
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.
560382 Rev. 1
Format Details
Alexa Fluor™ 647
Alexa Fluor™ 647 Dye is part of the BD red family of dyes. This is a small organic fluorochrome with an excitation maximum (Ex Max) at 653-nm and an emission maximum (Em Max) at 669-nm. Alexa Fluor 647 is designed to be excited by the Red laser (627-640 nm) and detected using an optical filter centered near 520-nm (e.g., a 660/20 nm bandpass filter). Please ensure that your instrument’s configurations (lasers and optical filters) are appropriate for this dye.
Alexa Fluor™ 647
Red 627-640 nm
653 nm
669 nm
560382 Rev.1
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
-
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: Immunofluorescence). 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). View Reference
560382 Rev. 1
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.
