Skip to main content Skip to navigation
Alexa Fluor® 647 Mouse Anti-Insulin
Alexa Fluor® 647 Mouse Anti-Insulin

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

LEFT: Untransfected (dashed line histogram) and human insulin-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-Insulin (Cat. No. 565689).

RIGHT: Mouse Beta-TC-6 cells (ATCC CRL-11506) 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 either Alexa Fluor® 647 Mouse IgG1, κ isotype control (Cat. No. 557783, dashed-line histogram) or Alexa Fluor® 647 Mouse Anti-Insulin (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 Canto™ II flow cytometry system.

Alexa Fluor® 647 Mouse Anti-Insulin

Fluorescent staining of insulin in human and mouse islets of Langerhans. Following antigen retrieval with BD Pharmingen™ Retrievagen A Buffer (Cat. No. 550524), sections from  formalin-fixed, paraffin-embedded sections of human (left) and mouse (right) pancreata were stained with Alexa Fluor® 647 Mouse Anti-Insulin (Cat. No. 565689, pseudocolored green). Cell nuclei were counterstained with DAPI (Cat. No. 564907, pseudocolored red). The images were captured using an ImageXpress® Micro XLS and merged using MetaXpress® Software (Molecular Devices, LLC).

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

LEFT: Untransfected (dashed line histogram) and human insulin-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-Insulin (Cat. No. 565689).

RIGHT: Mouse Beta-TC-6 cells (ATCC CRL-11506) 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 either Alexa Fluor® 647 Mouse IgG1, κ isotype control (Cat. No. 557783, dashed-line histogram) or Alexa Fluor® 647 Mouse Anti-Insulin (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 Canto™ II flow cytometry system.

Fluorescent staining of insulin in human and mouse islets of Langerhans. Following antigen retrieval with BD Pharmingen™ Retrievagen A Buffer (Cat. No. 550524), sections from  formalin-fixed, paraffin-embedded sections of human (left) and mouse (right) pancreata were stained with Alexa Fluor® 647 Mouse Anti-Insulin (Cat. No. 565689, pseudocolored green). Cell nuclei were counterstained with DAPI (Cat. No. 564907, pseudocolored red). The images were captured using an ImageXpress® Micro XLS and merged using MetaXpress® Software (Molecular Devices, LLC).

Product Details
Down Arrow Up Arrow


BD Pharmingen™
INS; IDDM; ILPR; IRDN; IDDM1; IDDM2; MODY10; Ins2
Human (QC Testing), Mouse, Rat (Tested in Development)
Mouse IgG1, κ
Mature human insulin (A- and B-chain) Recombinant Protein
Intracellular staining (flow cytometry) (Routinely Tested), Immunofluorescence, Immunohistochemistry-formalin (antigen retrieval required) (Tested During Development)
0.2 mg/ml
AB_2739331
Aqueous buffered solution containing protein stabilizer and ≤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 Alexa Fluor® 647 under optimum conditions, and unreacted Alexa Fluor® 647 was removed. Store undiluted at 4°C and protected from prolonged exposure to light. Do not freeze.

Product Notices

  1. Since applications vary, each investigator should titrate the reagent to obtain optimal results.
  2. Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
  3. 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.
  4. Alexa Fluor® is a registered trademark of Molecular Probes, Inc., Eugene, OR.
  5. Alexa Fluor® 647 fluorochrome emission is collected at the same instrument settings as for allophycocyanin (APC).
  6. All other brands are trademarks of their respective owners.
  7. For fluorochrome spectra and suitable instrument settings, please refer to our Multicolor Flow Cytometry web page at www.bdbiosciences.com/colors.
  8. 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.
  9. Species cross-reactivity detected in product development may not have been confirmed on every format and/or application.
  10. An isotype control should be used at the same concentration as the antibody of interest.
565689 Rev. 1
Antibody Details
Down Arrow Up Arrow
T56-706

The T56-706 monoclonal antibody specifically binds to insulin, a member of the insulin family of active peptides. 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. Insulin is produced by β cells in the islets of Langerhans of the pancreas. There, the precursor molecule, preproinsulin, is cleaved to proinsulin that is in turn cleaved to form the mature insulin hormone, which is composed of two peptides (A- and B-chains) linked by 2 disulfide bonds. Mature insulin is stored in granules in the β cells and is released to the blood in response to metabolic signals such as glucose, the amino acids arginine and leucine, and acetylcholine. Catecholamines can regulate blood glucose levels by either stimulating or inhibiting the release of insulin from β cells. The expression of insulin can be used to monitor the pancreatic differentiation of pluripotent stem cells.

565689 Rev. 1
Format Details
Down Arrow Up Arrow
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
565689 Rev.1
Citations & References
Down Arrow Up Arrow

Development References (5)

  1. Bell GI, Pictet RL, Rutter WJ, Cordell B, Tischer E, Goodman HM. Sequence of the human insulin gene. Nature. 1980; 284(5751):26-32. (Biology). View Reference
  2. 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
  3. 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
  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
View All (5) View Less
565689 Rev. 1

Please refer to Support Documents for Quality Certificates


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.