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Alexa Fluor® 647 Mouse Anti-C-Peptide
Alexa Fluor® 647 Mouse Anti-C-Peptide

Flow cytometric analysis of C-Peptide expression in human INS-transfected 293F cells and a mouse insulinoma cell line.

LEFT: Untransfected (dashed-line histogram) and human INS-transfected (solid-line histogram) 293F 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 then washed and stained with Alexa Fluor® 647 Mouse Anti-C-Peptide (Cat. No. 565831).

RIGHT: Beta-TC-6 cells (ATCC CRL-11505) were fixed with BD Cytofix™ Fixation Buffer (Cat. No. 554655) and permeabilized with BD Phosflow™ Perm Buffer III (Cat. No. 558050). The cells were washed and stained with either Alexa Fluor® 647 Mouse IgG1 κ Isotype control (Cat. No. 557783, dashed-line histogram) or Alexa Fluor® 647 Mouse Anti-C-Peptide (Cat. No. 565831, solid-line histogram).

All fluorescence histograms were derived from gated events with the forward and side light-scatter characteristics of intact cells. Flow cytometry was performed on a BD FACSCanto™ II flow cytometry system.

Alexa Fluor® 647 Mouse Anti-C-Peptide

Immunohistofluorescent staining of C-Peptide in human, rat, and mouse islets of Langerhans. Following antigen retrieval with BD Retrievagen A Buffer (Cat. No. 550524), sections from formalin-fixed, paraffin-embedded human (top panel), rat (center panel), and mouse (bottom panel) pancreata were blocked using an Avidin/ Biotin Blocking Kit (Vector Laboratories, Cat. No. SP-2001) as recommended by the manufacturer. The sections were then stained with Alexa Fluor® 647 Mouse Anti-C-Peptide (Cat. No. 565831, pseudocolored green). Cell nuclei were counterstained with DAPI (Cat. No. 564907, pseudocolored red). The photographs were performed on a standard  epifluorescence microscope. Original magnifications: 20X.

Flow cytometric analysis of C-Peptide expression in human INS-transfected 293F cells and a mouse insulinoma cell line.

LEFT: Untransfected (dashed-line histogram) and human INS-transfected (solid-line histogram) 293F 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 then washed and stained with Alexa Fluor® 647 Mouse Anti-C-Peptide (Cat. No. 565831).

RIGHT: Beta-TC-6 cells (ATCC CRL-11505) were fixed with BD Cytofix™ Fixation Buffer (Cat. No. 554655) and permeabilized with BD Phosflow™ Perm Buffer III (Cat. No. 558050). The cells were washed and stained with either Alexa Fluor® 647 Mouse IgG1 κ Isotype control (Cat. No. 557783, dashed-line histogram) or Alexa Fluor® 647 Mouse Anti-C-Peptide (Cat. No. 565831, solid-line histogram).

All fluorescence histograms were derived from gated events with the forward and side light-scatter characteristics of intact cells. Flow cytometry was performed on a BD FACSCanto™ II flow cytometry system.

Immunohistofluorescent staining of C-Peptide in human, rat, and mouse islets of Langerhans. Following antigen retrieval with BD Retrievagen A Buffer (Cat. No. 550524), sections from formalin-fixed, paraffin-embedded human (top panel), rat (center panel), and mouse (bottom panel) pancreata were blocked using an Avidin/ Biotin Blocking Kit (Vector Laboratories, Cat. No. SP-2001) as recommended by the manufacturer. The sections were then stained with Alexa Fluor® 647 Mouse Anti-C-Peptide (Cat. No. 565831, pseudocolored green). Cell nuclei were counterstained with DAPI (Cat. No. 564907, pseudocolored red). The photographs were performed on a standard  epifluorescence microscope. Original magnifications: 20X.

Product Details
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BD Pharmingen™
Human (QC Testing), Mouse, Rat (Tested in Development)
Mouse BALB/c IgG1, κ
Human C-Peptide
Intracellular staining (flow cytometry) (Routinely Tested), Immunofluorescence, Immunohistochemistry-paraffin (Tested During Development)
0.2 mg/ml
AB_2739371
Aqueous buffered solution containing 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. Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
  2. 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.
  3. Alexa Fluor® is a registered trademark of Molecular Probes, Inc., Eugene, OR.
  4. Alexa Fluor® 647 fluorochrome emission is collected at the same instrument settings as for allophycocyanin (APC).
  5. 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.
  6. For fluorochrome spectra and suitable instrument settings, please refer to our Multicolor Flow Cytometry web page at www.bdbiosciences.com/colors.
  7. Since applications vary, each investigator should titrate the reagent to obtain optimal results.
  8. An isotype control should be used at the same concentration as the antibody of interest.
565831 Rev. 1
Antibody Details
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U8-424

The U8-424 monoclonal antibody specifically binds to human, mouse, and rat C-Peptide, the connecting peptide that links the A- and B-chains in the proinsulin molecule. The A- and B- chains and C-Peptide are encoded by the transcript of the INS gene and are produced by the β cells in the islets of Langerhans of the pancreas. As the biosynthesis of insulin proceeds, the C-Peptide is cleaved from proinsulin to form the mature insulin hormone, which is composed of the A- and B-chains linked by 2 disulfide bonds. Mature insulin and C-Peptide are stored in granules in the β cells and are released to the blood in response to metabolic signals such as glucose, the amino acids arginine and leucine, and acetylcholine. As a result, C-Peptide is released into the blood stream in an equimolar amount to insulin; the serum level of C-Peptide correlates with pancreatic β cell function and the amount of insulin being produced. The expression of C-Peptide can be used to monitor the pancreatic differentiation of pluripotent stem cells. Insulin is an evolutionarily conserved peptide hormone that binds to receptors on target cells (primarily adipose and muscle) to promote the absorption of glucose from the blood, thus regulating fat and carbohydrate metabolism. C-Peptide itself binds to many cell types, independently of the insulin receptor, and initiates several intracellular signaling cascades.

565831 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
565831 Rev.1
Citations & References
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Development References (7)

  1. D'Amour KA, Bang AG, Eliazer S, et al . Production of pancreatic hormone-expressing endocrine cells from human embryonic stem cells. Nat Biotechnol. 2006; 24(12):1481-1483. (Biology). View Reference
  2. Kelly OG, Chan MY, Martinson LA, et al. Cell-surface markers for the isolation of pancreatic cell types derived from human embryonic stem cells. Nat Biotechnol. 2011; 29(8):750-756. (Biology). View Reference
  3. Ko AS, Smyth DG, Marktussen J, Sundby F. The amino acid sequence of the C-peptide of human proinsulin.. Eur J Biochem. 1971; 20(2):190-9. (Biology). View Reference
  4. Pagliuca FW, Millman JR, Gürtler M, et al. Generation of functional human pancreatic β cells in vitro. Cell. 2014; 159(2):428-439. (Biology). View Reference
  5. Rezania A, Bruin JE, Riedel MJ et al. Maturation of human embryonic stem cell-derived pancreatic progenitors into functional islets capable of treating pre-existing diabetes in mice. Diabetes. 2012; 61(8):2016-2029. (Biology). View Reference
  6. Suckale J, Solimena M. The insulin secretory granule as a signaling hub.. Trends Endocrinol Metab. 2010; 21(10):599-609. (Biology). View Reference
  7. Yosten GL, Kolar GR. The Physiology of Proinsulin C-Peptide: Unanswered Questions and a Proposed Model.. Physiology (Bethesda). 2015; 30(4):327-32. (Biology). View Reference
View All (7) View Less
565831 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.