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Alexa Fluor® 647 Mouse Anti-Human Lin-28
Alexa Fluor® 647 Mouse Anti-Human Lin-28
Flow cytometric analysis of Lin-28 expression in human embryonic stem (ES) cells. H9 human ES cells (WiCell, Madison, WI) passage 47 were grown in mTeSR™1 Medium (StemCell Technologies) on Matrigel® hESC-Qualified Matrix (Corning). They were harvested with BD™ Accutase™ Cell Detachment Solution (Cat. No. 561527), fixed with BD Cytofix™ Fixation Buffer (Cat. No. 554655), and permeabilized with BD Phosflow™ Perm Buffer III (Cat. No. 558050). The cells were then stained with either Alexa Fluor® 647 Mouse IgG1, κ Isotype Control (dashed line, Cat. No. 557714) or Alexa Fluor® 647 Mouse Anti-Human Lin-28 monoclonal antibody (solid line, Cat. No. 564280) at matched concentrations. The histograms were derived from gated events based on light scattering characteristics of H9 human ES cells. Flow cytometry was performed on a BD LSRFortessa™ Flow Cytometry System.
Alexa Fluor® 647 Mouse Anti-Human Lin-28
Immunofluorescent staining of human embryonic stem (ES) cell line. H9 cells (WiCell, Madison, WI) passage 49 were grown in mTeSR™1 Medium (StemCell Technologies) on Matrigel® hESC-qualified matrix (Corning) in a Falcon® 96 Well Imaging Plate (Corning). They were fixed with BD Cytofix™ Fixation Buffer (Cat. No. 554655), permeabilized using BD Perm/Wash™ Perm/Wash Buffer (Cat. No. 554723), and stained with 20 µl of Alexa Fluor® 647 Mouse Anti-Human Lin-28 monoclonal antibody (pseudo-colored red) per 50 µl of stain solution per well. The cell nuclei were counterstained with DAPI (pseudo-colored blue). The images were captured on a BD Pathway™ 435 Cell Analyzer and merged using BD Attovision™ software. BD Phosflow™ Perm Buffer III (Cat. No. 558050) is also suitable for permeabilization.
Flow cytometric analysis of Lin-28 expression in human embryonic stem (ES) cells. H9 human ES cells (WiCell, Madison, WI) passage 47 were grown in mTeSR™1 Medium (StemCell Technologies) on Matrigel® hESC-Qualified Matrix (Corning). They were harvested with BD™ Accutase™ Cell Detachment Solution (Cat. No. 561527), fixed with BD Cytofix™ Fixation Buffer (Cat. No. 554655), and permeabilized with BD Phosflow™ Perm Buffer III (Cat. No. 558050). The cells were then stained with either Alexa Fluor® 647 Mouse IgG1, κ Isotype Control (dashed line, Cat. No. 557714) or Alexa Fluor® 647 Mouse Anti-Human Lin-28 monoclonal antibody (solid line, Cat. No. 564280) at matched concentrations. The histograms were derived from gated events based on light scattering characteristics of H9 human ES cells. Flow cytometry was performed on a BD LSRFortessa™ Flow Cytometry System.
Immunofluorescent staining of human embryonic stem (ES) cell line. H9 cells (WiCell, Madison, WI) passage 49 were grown in mTeSR™1 Medium (StemCell Technologies) on Matrigel® hESC-qualified matrix (Corning) in a Falcon® 96 Well Imaging Plate (Corning). They were fixed with BD Cytofix™ Fixation Buffer (Cat. No. 554655), permeabilized using BD Perm/Wash™ Perm/Wash Buffer (Cat. No. 554723), and stained with 20 µl of Alexa Fluor® 647 Mouse Anti-Human Lin-28 monoclonal antibody (pseudo-colored red) per 50 µl of stain solution per well. The cell nuclei were counterstained with DAPI (pseudo-colored blue). The images were captured on a BD Pathway™ 435 Cell Analyzer and merged using BD Attovision™ software. BD Phosflow™ Perm Buffer III (Cat. No. 558050) is also suitable for permeabilization.
Product Details
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BD Pharmingen™
Lin-28A, LIN28A, CSDD1, LIN28, ZCCHC1
Human (QC Testing)
Mouse IgG1, κ
Human Lin-28 Recombinant Protein
Intracellular staining (flow cytometry) (Routinely Tested), Bioimaging, Immunofluorescence (Tested During Development)
5 µl
AB_2869558
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® 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. 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.
  4. Source of all serum proteins is from USDA inspected abattoirs located in the United States.
  5. Alexa Fluor® 647 fluorochrome emission is collected at the same instrument settings as for allophycocyanin (APC).
  6. 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.
  7. Alexa Fluor® is a registered trademark of Molecular Probes, Inc., Eugene, OR.
  8. mTESR™1 is a trademark of StemCell Technologies.
  9. Accutase is a registered trademark of Innovative Cell Technologies, Inc.
  10. For fluorochrome spectra and suitable instrument settings, please refer to our Multicolor Flow Cytometry web page at www.bdbiosciences.com/colors.
  11. Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
564280 Rev. 1
Antibody Details
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6D1F9

Lin-28 is an evolutionarily conserved RNA-binding protein that is expressed in embryonic tissue, embryonic stem cells, and some adult tissues (e.g. cardiac and skeletal muscle). Within these cells, it is localized to ribosomes, P-bodies, and stress granules. During embryonic development, Lin-28 acts as a "translational enhancer" by binding to certain mRNAs and increasing their translation efficiency. Additionally, Lin-28 is overexpressed in some human cancer cell lines and is associated with the malignant transformation of cells as well as advanced human malignancies. This role is thought to be tied to the binding of Lin-28 to pre-microRNA of let-7, which in its mature form promotes terminal differentiation of stem cells and suppresses tumors. The binding of Lin-28 to the immature version of let-7 blocks its cleavage into mature microRNA, thereby preventing it from carrying out its function in tumor suppression. Lin-28, in combination with additional factors, has been used to reprogram human somatic cells into induced pluripotent stem cells.

564280 Rev. 1
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
564280 Rev.1
Citations & References
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Development References (7)

  1. Balzer E, Moss EG. Localization of the developmental timing regulator Lin28 to mRNP complexes, P-bodies and stress granules. RNA Biol. 2007; 4(1):16-25. (Biology). View Reference
  2. Darr H, Benvenisty N. Genetic analysis of the role of the reprogramming gene LIN-28 in human embryonic stem cells. Stem Cells. 2008. (Biology). View Reference
  3. Newman MA, Thomson JM, Hammond SM. Lin-28 interaction with the Let-7 precursor loop mediates regulated microRNA processing. RNA. 2008; 14(8):1539-1549. (Biology). View Reference
  4. Polesskaya A, Cuvellier S, Naguibneva I, Duquet A, Moss EG, Harel-Bellan A. Lin-28 binds IGF-2 mRNA and participates in skeletal myogenesis by increasing translation efficiency. Genes Dev. 2007; 21(9):1125-1138. (Biology). View Reference
  5. Viswanathan SR, Daley GQ, Gregory RI. Selective blockade of microRNA processing by Lin28. Science. 2008; 320:97-100. (Biology: Array). View Reference
  6. Viswanathan SR, Powers JT, Einhorn W, et al. Lin28 promotes transformation and is associated with advanced human malignancies. Nat Genet. 2009; 41(7):843-848. (Biology). View Reference
  7. Yu J, Vodyanik MA, Smuga-Otto K, et al. Induced pluripotent stem cell lines derived from human somatic cells. Science. 2007; 318(5858):1917-1920. (Biology). View Reference
View All (7) View Less
564280 Rev. 1

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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.