Alexa Fluor® 647 Mouse Anti-C-Peptide

BD Pharmingen™ Alexa Fluor® 647 Mouse Anti-C-Peptide

Name: Alexa Fluor® 647 Mouse Anti-C-Peptide
Brand: Alexa Fluor® 647 Mouse Anti-C-Peptide
SKU: 565831

Clone U8-424 (RUO)

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.

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

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.

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Catalog No:565831

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Product Details

Brand: BD Pharmingen™

Reactivity: Human (QC Testing), Mouse, Rat (Tested in Development)

Isotype: Mouse BALB/c IgG1, κ

Immunogen: Human C-Peptide

Application: Intracellular staining (flow cytometry) (Routinely Tested), Immunofluorescence, Immunohistochemistry-paraffin (Tested During Development)

Concentration: 0.2 mg/ml

RRID: AB_2739371

Storage Buffer: Aqueous buffered solution containing protein stabilizer and ≤0.09% sodium azide.

Regulatory Status: 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

Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
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.
Alexa Fluor® 647 fluorochrome emission is collected at the same instrument settings as for allophycocyanin (APC).
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.
For fluorochrome spectra and suitable instrument settings, please refer to our Multicolor Flow Cytometry web page at www.bdbiosciences.com/colors.
Since applications vary, each investigator should titrate the reagent to obtain optimal results.
An isotype control should be used at the same concentration as the antibody of interest.

Data Sheets

Technical Data Sheet (TDS)

Safety Data Sheet

Companion Products

Stain Buffer (BSA) RUO

Size 500 ML Cat No. 554657

Stain Buffer (FBS) RUO

Size 500 ML Cat No. 554656

Fixation Buffer RUO

Size 100 ML Cat No. 554655

Perm Buffer III RUO

Size 125 ML Cat No. 558050

Retrievagen A (pH 6.0) RUO

Size 1000 ML Cat No. 550524

Alexa Fluor® 647 Mouse IgG1 κ Isotype control RUO

Size 50 Tests Cat No. 557783

View Details

565831 Rev. 1

Antibody Details

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

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 670-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.

Format: Alexa Fluor™ 647

Excitation Max: 653 nm

Emission Max: 669 nm

565831 Rev.1

Citations & References

View Development References

Development References (7)

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
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
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
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
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
Suckale J, Solimena M. The insulin secretory granule as a signaling hub.. Trends Endocrinol Metab. 2010; 21(10):599-609. (Biology). View Reference
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)

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.