Flow cytometric analysis of PDX-1 in mouse pancreatic tumor (insulinoma) cells. Beta-TC-6 (ATCC CRL-11506) 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 stained with either PerCP-Cy™5.5 Mouse IgG1, isotype control (Cat. No. 550795; dashed line histogram) or PerCP-Cy™5.5 Mouse Anti-PDX-1 monoclonal antibody (Cat. No. 563436, solid line histogram) at matched concentrations. The fluorescence histograms were derived from events with the forward and side light-scatter characteristics of intact cells. Flow cytometric analysis was performed using a BD™ LSR II Flow Cytometer System. This antibody has been found to be reactive with both mouse and human PDX-1, and it can also be used on fixed cells that were permeabilized with BD Phosflow™ Perm/Wash Buffer I (Cat. No. 557885).
Flow cytometric analysis of PDX-1 in mouse pancreatic tumor (insulinoma) cells. Beta-TC-6 (ATCC CRL-11506) 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 stained with either PerCP-Cy™5.5 Mouse IgG1, isotype control (Cat. No. 550795; dashed line histogram) or PerCP-Cy™5.5 Mouse Anti-PDX-1 monoclonal antibody (Cat. No. 563436, solid line histogram) at matched concentrations. The fluorescence histograms were derived from events with the forward and side light-scatter characteristics of intact cells. Flow cytometric analysis was performed using a BD™ LSR II Flow Cytometer System. This antibody has been found to be reactive with both mouse and human PDX-1, and it can also be used on fixed cells that were permeabilized with BD Phosflow™ Perm/Wash Buffer I (Cat. No. 557885).
Product Details
BD Pharmingen™
PDX1, GSF, IPF-1, IPF1, IUF-1, IDX-1, STF-1
Mouse (QC Testing), Human (Tested in Development)
Mouse IgG1, κ
Human PDX-1 Recombinant Protein
Intracellular staining (flow cytometry) (Routinely Tested)
5 µl
3651
AB_2738204
Aqueous buffered solution containing BSA 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 with PerCP-Cy5.5 under optimum conditions, and unconjugated antibody and free PerCP-Cy5.5 were removed. Storage of PerCP-Cy5.5 conjugates in unoptimized diluent is not recommended and may result in loss of signal intensity.
Product Notices
- 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).
- An isotype control should be used at the same concentration as the antibody of interest.
- Source of all serum proteins is from USDA inspected abattoirs located in the United States.
- 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.
- Species cross-reactivity detected in product development may not have been confirmed on every format and/or application.
- Please observe the following precautions: Absorption of visible light can significantly alter the energy transfer occurring in any tandem fluorochrome conjugate; therefore, we recommend that special precautions be taken (such as wrapping vials, tubes, or racks in aluminum foil) to prevent exposure of conjugated reagents, including cells stained with those reagents, to room illumination.
- PerCP-Cy5.5–labelled antibodies can be used with FITC- and R-PE–labelled reagents in single-laser flow cytometers with no significant spectral overlap of PerCP-Cy5.5, FITC, and R-PE fluorescence.
- PerCP-Cy5.5 is optimized for use with a single argon ion laser emitting 488-nm light. Because of the broad absorption spectrum of the tandem fluorochrome, extra care must be taken when using dual-laser cytometers, which may directly excite both PerCP and Cy5.5™. We recommend the use of cross-beam compensation during data acquisition or software compensation during data analysis.
- For fluorochrome spectra and suitable instrument settings, please refer to our Multicolor Flow Cytometry web page at www.bdbiosciences.com/colors.
- Cy is a trademark of GE Healthcare.
- Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
Companion Products
Stain Buffer (FBS) RUO
Size
500 mL
Cat No.
554656
Stain Buffer (BSA) RUO
Size
500 mL
Cat No.
554657
Fixation Buffer RUO
Size
100 mL
Cat No.
554655
Perm Buffer III RUO
Size
125 mL
Cat No.
558050
Perm/Wash Buffer I RUO
Size
125 mL
Cat No.
557885
PerCP-Cy™5.5 Mouse IgG1 κ Isotype Control RUO
Size
0.1 mg
Cat No.
550795
563436 Rev. 2
Antibody Details
658A5
The 658A5 monoclonal antibody binds PDX-1 or Pancreas/Duodenum Homeobox Protein-1 encoded by the PDX1 or IPF1 (Insulin Promoter Factor 1) gene. PDX-1 is a key regulator of pancreatic development and adult beta-cell function. Loss of PDX1 gene function in mice and humans results in pancreatic agenesis. PDX-1 is also involved in endocrine precursor cell development by binding the DNA-binding transcription factor Neurogenin-3. PDX-1-positive cells have been obtained by the directed differentiation of human embryonic stem cells through a definitive endoderm lineage.
563436 Rev. 2
Format Details
PerCP-Cy5.5
PerCP-Cy5.5 dye is part of the BD blue family of dyes. This tandem fluorochrome is comprised of a fluorescent protein complex (PerCP) with an excitation maximum (Ex Max) of 482 nm and an acceptor dye with an emission maximum (Em Max) at 676 nm. PerCP-Cy5 is designed to be excited by the blue laser (488-nm) and detected using an optical filter centered near 680 nm (e.g., a 695/40 nm bandpass filter). The donor dye can be partially excited by the Violet (405-nm) laser resulting in cross-laser excitation and fluorescence spillover. Please ensure that your instrument’s configurations (lasers and optical filters) are appropriate for this dye.
PerCP-Cy5.5
Blue 488 nm
482 nm
676 nm
563436 Rev.2
Citations & References
Development References (5)
-
Babu DA, Deering TG, Mirmira RG. A feat of metabolic proportions: Pdx1 orchestrates islet development and function in the maintenance of glucose homeostasis. Mol Genet Metab. 2007; 92(1-2):43-55. (Biology). View Reference
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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: Calcium Flux). View Reference
-
Gannon M, Ables ET, Crawford L, et al. pdx-1 function is specifically required in embryonic beta cells to generate appropriate numbers of endocrine cell types and maintain glucose homeostasis. Dev Biol. 2008; 314(2):406-417. (Biology). View Reference
-
Gu G, Dubauskaite J, Melton DA. Direct evidence for the pancreatic lineage: NGN3+ cells are islet progenitors and are distinct from duct progenitors. Development. 2002; 129(10):2447-2457. (Biology). View Reference
-
Ohlsson H, Karlsson K, Edlund T. IPF1, a homeodomain-containing transactivator of the insulin gene. EMBO J. 1993; 12(11):4251-4259. (Biology). View Reference
563436 Rev. 2
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.
