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Alexa Fluor® 647 Mouse Anti-Human CD171
Alexa Fluor® 647 Mouse Anti-Human CD171

Flow cytometric analysis of CD171 expression on human M21 melanoma cells. Cells from the human M21 melanoma cell line were stained with either Alexa Fluor® 647 Mouse IgG2a, κ Isotype Control (Cat. No. 557715; dashed line histogram) or Alexa Fluor® 647 Mouse Anti-Human CD171 antibody (Cat. No. 564194; solid line histogram). The fluorescence histograms were derived from gated events with the forward and side light-scatter characteristics of viable M21 cells. Flow cytometric analysis was performed using a BD™ LSR II Flow Cytometer System.

Flow cytometric analysis of CD171 expression on human M21 melanoma cells. Cells from the human M21 melanoma cell line were stained with either Alexa Fluor® 647 Mouse IgG2a, κ Isotype Control (Cat. No. 557715; dashed line histogram) or Alexa Fluor® 647 Mouse Anti-Human CD171 antibody (Cat. No. 564194; solid line histogram). The fluorescence histograms were derived from gated events with the forward and side light-scatter characteristics of viable M21 cells. Flow cytometric analysis was performed using a BD™ LSR II Flow Cytometer System.

Product Details
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BD Pharmingen™
L1 Neurite Cell Adhesion Molecule; N-CAM-L1; L1CAM ; CAML1; MIC5
Human (QC Testing)
Mouse IgG2a
Human SK-N-AS Neuroblastoma Cell Line
Flow cytometry (Routinely Tested)
215/200 kDa
5 µl
VII 70700
AB_2738661
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 to Alexa Fluor® 647 under optimum conditions, and unreacted Alexa Fluor® 647 was removed.

Product Notices

  1. 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).
  2. An isotype control should be used at the same concentration as the antibody of interest.
  3. Source of all serum proteins is from USDA inspected abattoirs located in the United States.
  4. 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.
  5. 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.
  6. Alexa Fluor® is a registered trademark of Molecular Probes, Inc., Eugene, OR.
  7. Alexa Fluor® 647 fluorochrome emission is collected at the same instrument settings as for allophycocyanin (APC).
  8. For fluorochrome spectra and suitable instrument settings, please refer to our Multicolor Flow Cytometry web page at www.bdbiosciences.com/colors.
  9. Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
564194 Rev. 2
Antibody Details
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5G3

Neurite adhesion molecule L1 has been implicated in neuron-neuron and neuron-Schwann cell adhesion in vertebrates. L1-like molecules, found in mouse, rat, chicken, and human, promote axonal elongation and may also play a role in regeneration of axons after injury. Molecular cloning data suggest 87% amino acid identity between mouse and human L1 molecules. 5G3 antigen (Ag), originally defined by monoclonal antibody 5G3, is considered to be the human homologue of mouse L1. The 5G3 antibody was developed against a human neuroblastoma cell line to use as a probe for the elucidating the biological characteristics of neuroblastoma. 5G3 specifically recognizes a neuroblastoma target glycoprotein antigen of 215 kDa and its 200 kDa precursor. The 215 kDa molecule is expressed on the cell surface; whereas the 200 kDa precursor is shed from the cell surface. The 215 and 200 kDa species also differ in their posttranslational modification patterns. The 5G3 antibody has been used as a marker for neuroblastoma, and to purify 5G3 Ag from normal adult human brain.

The antibody recognizes human L1 on human neuroblastoma cell lines and tissues. Reactivity has been tested on a variety of malignant and normal tissues. Squamous lung, squamous skin, and osteogenic sarcoma cell lines were positive, as were two out of eight melanoma cell lines tested. A variety of other cell lines and tumor tissues tested negative. 5G3 did not react with either T or B lymphoblastoid cell lines or a fibroblast cell line. Among all the normal tissues tested, mAb 5G3 reacted only with cerebellum.

The molecular masses observed using mAb 5G3 may vary among immunoprecipitation isolates. In normal human cerebellum, 5G3 Ag migrated as a 190/200 kDa doublet, 140 kDa band with minor bands at 80 and 65 kDa. 5G3 Ag isolated from SK-N-AS cells migrates as 200 to 215 kDa bands, or as a diffuse band ranging from 200 to 215 kDa. Additional bands have been described at 140 to 150 kDa in SK-N-AS cells. Only the 200 kDa band has been detected in culture media from SK-N-AS cells.

564194 Rev. 2
Format Details
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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
564194 Rev.2
Citations & References
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Development References (12)

  1. Cheresh DA, Honsik CJ, Staffileno LK, Jung G, Reisfeld RA. Disialoganglioside GD3 on human melanoma serves as a relevant target antigen for monoclonal antibody-mediated tumor cytolysis. Proc Natl Acad Sci U S A. 1985; 82(15):5155-5159. (Biology). View Reference
  2. Lemmon V, McLoon SC. The appearance of an L1-like molecule in the chick primary visual pathway. J Neurosci. 1986; 6(10):2987-2994. (Biology). View Reference
  3. Mechtersheimer S, Gutwein P, Agmon-Levin N, et al. Ectodomain shedding of L1 adhesion molecule promotes cell migration by autocrine binding to integrins. J Cell Biol. 2001; 155(4):661-673. (Clone-specific: Western blot). View Reference
  4. Mujoo K, Spiro RC, Reisfeld RA. Characterization of a unique glycoprotein antigen expressed on the surface of human neuroblastoma cells. J Biol Chem. 1986; 261(22):10299-10305. (Immunogen: Immunofluorescence, Immunohistochemistry, Western blot). View Reference
  5. Nayeem N, Silletti S, Yang X, et al. A potential role for the plasmin(ogen) system in the posttranslational cleavage of the neural cell adhesion molecule L1. J Cell Sci. 1999; 112(24):4739-4749. (Clone-specific: Western blot). View Reference
  6. Pancook JD, Reisfeld RA, Varki N, Vitiello A, Fox RI, Montgomery AM. Expression and regulation of the neural cell adhesion molecule L1 on human cells of myelomonocytic and lymphoid origin.. J Immunol. 1997; 158(9):4413-21. (Biology). View Reference
  7. Rathjen FG, Schachner M. Immunocytological and biochemical characterization of a new neuronal cell surface component (L1 antigen) which is involved in cell adhesion. EMBO J. 1984; 3(1):1-10. (Biology). View Reference
  8. Rathjen FG, Wolff JM, Chang S, Bonhoeffer F, Raper JA. Neurofascin: a novel chick cell-surface glycoprotein involved in neurite-neurite interactions. Cell. 1987; 51(5):841-849. (Biology). View Reference
  9. Reid RA, Hemperly JJ. Variants of human L1 cell adhesion molecule arise through alternate splicing of RNA. J Mol Neurosci. 1992; 3(3):127-135. (Biology). View Reference
  10. Salton SR, Shelanski ML, Greene LA. Biochemical properties of the nerve growth factor-inducible large external (NILE) glycoprotein. J Neurosci. 1983; 3(12):2420-2430. (Biology). View Reference
  11. Wolff JM, Frank R, Mujoo K, Spiro RC, Reisfeld RA, Rathjen FG. A human brain glycoprotein related to the mouse cell adhesion molecule L1.. J Biol Chem. 1988; 263(24):11943-7. (Biology). View Reference
  12. Wolff R, Plow EF, Ginsberg MH. Interaction of thrombospondin with resting and stimulated human platelets. J Biol Chem. 1986; 261(15):6840-6846. (Clone-specific: Western blot). View Reference
View All (12) View Less
564194 Rev. 2

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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.