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Alexa Fluor™ 488 Mouse Anti-C-Peptide
Alexa Fluor™ 488 Mouse Anti-C-Peptide
Flow cytometric analysis of C-Peptide expression in Human Insulin-transfected 293F cells and a Mouse insulinoma cell line.    Left Plot - Human 293F untransfected (dashed line histogram) and Insulin-transfected (solid line histogram) 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™ 488 Mouse Anti-C-Peptide antibody (Cat. No. 570993/571064; solid line histogram) at 0.5 µg/test.    Right Plot - Cells from the Mouse Beta-TC-6 (Insulinoma, ATCC© CRL-3605™) cell line were fixed with BD Cytofix™ Fixation Buffer and permeabilized with BD Phosflow™ Perm Buffer III. The cells were washed and stained with either Alexa Fluor™ 488 Mouse IgG1, κ Isotype Control (Cat. No. 565572; dashed line histogram) or Alexa Fluor™ 488 Mouse Anti-C-Peptide antibody (solid line histogram) at 0.5 µg/test. The fluorescence histograms showing C-Peptide expression were derived from gated events with the forward and side light-scatter characteristics of intact cells.    Flow cytometry and data analysis were performed using a BD FACSCanto™ II Flow Cytometry System and FlowJo™ Software. Data shown on this Technical Data Sheet is not lot specific.
Flow cytometric analysis of C-Peptide expression in Human Insulin-transfected 293F cells and a Mouse insulinoma cell line.    Left Plot - Human 293F untransfected (dashed line histogram) and Insulin-transfected (solid line histogram) 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™ 488 Mouse Anti-C-Peptide antibody (Cat. No. 570993/571064; solid line histogram) at 0.5 µg/test.    Right Plot - Cells from the Mouse Beta-TC-6 (Insulinoma, ATCC© CRL-3605™) cell line were fixed with BD Cytofix™ Fixation Buffer and permeabilized with BD Phosflow™ Perm Buffer III. The cells were washed and stained with either Alexa Fluor™ 488 Mouse IgG1, κ Isotype Control (Cat. No. 565572; dashed line histogram) or Alexa Fluor™ 488 Mouse Anti-C-Peptide antibody (solid line histogram) at 0.5 µg/test. The fluorescence histograms showing C-Peptide expression were derived from gated events with the forward and side light-scatter characteristics of intact cells.    Flow cytometry and data analysis were performed using a BD FACSCanto™ II Flow Cytometry System and FlowJo™ Software. Data shown on this Technical Data Sheet is not lot specific.
Product Details
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BD Pharmingen™
INS; insulin; preproinsulin; proinsulin
Human (QC Testing), Mouse,Rat (Tested in Development)
Mouse BALB/c IgG1, κ
Human C-Peptide
Intracellular staining (flow cytometry) (Routinely Tested)
0.2 mg/ml
3630, 16334, 24506
Aqueous buffered solution containing ≤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 the dye under optimum conditions and unreacted dye was removed. Store undiluted at 4°C and protected from prolonged exposure to light. Do not freeze.

Recommended Assay Procedures

BD® CompBeads can be used as surrogates to assess fluorescence spillover (compensation).  When fluorochrome conjugated antibodies are bound to BD® CompBeads, they have spectral properties very similar to cells.  However, for some fluorochromes there can be small differences in spectral emissions compared to cells, resulting in spillover values that differ when compared to biological controls.  It is strongly recommended that when using a reagent for the first time, users compare the spillover on cells and BD® CompBeads to ensure that BD® CompBeads are appropriate for your specific cellular application.

Product Notices

  1. Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
  2. Alexa Fluor® 488 fluorochrome emission is collected at the same instrument settings as for fluorescein isothiocyanate (FITC).
  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. Since applications vary, each investigator should titrate the reagent to obtain optimal results.
  5. For fluorochrome spectra and suitable instrument settings, please refer to our Multicolor Flow Cytometry web page at www.bdbiosciences.com/colors.
  6. An isotype control should be used at the same concentration as the antibody of interest.
  7. Species cross-reactivity detected in product development may not have been confirmed on every format and/or application.
  8. This product is provided under an intellectual property license between Life Technologies Corporation and BD Businesses. The purchase of this product conveys to the buyer the non-transferable right to use the purchased amount of the product and components of the product in research conducted by the buyer (whether the buyer is an academic or for-profit entity). The buyer cannot sell or otherwise transfer (a) this product (b) its components or (c) materials made using this product or its components to a third party or otherwise use this product or its components or materials made using this product or its components for Commercial Purposes. Commercial Purposes means any activity by a party for consideration and may include, but is not limited to: (1) use of the product or its components in manufacturing; (2) use of the product or its components to provide a service, information, or data; (3) use of the product or its components for therapeutic, diagnostic or prophylactic purposes; or (4) resale of the product or its components, whether or not such product or its components are resold for use in research. For information on purchasing a license to this product for any other use, contact Life Technologies Corporation, Cell Analysis Business Unit Business Development, 29851 Willow Creek Road, Eugene, OR 97402, USA, Tel: (541) 465-8300. Fax: (541) 335-0504.
  9. Please refer to http://regdocs.bd.com to access safety data sheets (SDS).
  10. Alexa Fluor™ is a trademark of Life Technologies Corporation.
  11. For U.S. patents that may apply, see bd.com/patents.
571064 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.

571064 Rev. 1
Format Details
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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
571064 Rev.1
Citations & References
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View product citations for antibody "571064" on CiteAb

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
571064 Rev. 1

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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.