BD Horizon™ RB780 Mouse Anti-Human FoxA2
Clone N17-280 (RUO)
Flow cytometric analysis of FoxA2 expression in Hep G2 cells. Cells from the Human Hep G2 [HEPG2] (Hepatocellular carcinoma, ATCC® HB-8065™) cell line were fixed (10 min, 37ºC) with pre-warmed BD Cytofix™ Buffer (Cat. No. 554655) and then permeabilized (on ice, 30 min) with BD Phosflow™ Perm Buffer III (Cat. No. 558050). The cells were washed with BD Pharmingen™ Stain Buffer (FBS) [Cat. No. 554656] and stained with either BD Horizon™ RB780 Mouse IgG1, κ Isotype Control (Cat. No. 568532; dashed line histogram) or BD Horizon™ RB780 Mouse Anti-Human FoxA2 antibody (Cat. No. 571077/571080; solid line histogram). The fluorescence histogram showing FoxA2 expression (or Ig Isotype control staining) was 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 FACSymphony™ A5 Cell Analyzer System and FlowJo™ Software.
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Flow cytometric analysis of FoxA2 expression in Hep G2 cells. Cells from the Human Hep G2 [HEPG2] (Hepatocellular carcinoma, ATCC® HB-8065™) cell line were fixed (10 min, 37ºC) with pre-warmed BD Cytofix™ Buffer (Cat. No. 554655) and then permeabilized (on ice, 30 min) with BD Phosflow™ Perm Buffer III (Cat. No. 558050). The cells were washed with BD Pharmingen™ Stain Buffer (FBS) [Cat. No. 554656] and stained with either BD Horizon™ RB780 Mouse IgG1, κ Isotype Control (Cat. No. 568532; dashed line histogram) or BD Horizon™ RB780 Mouse Anti-Human FoxA2 antibody (Cat. No. 571077/571080; solid line histogram). The fluorescence histogram showing FoxA2 expression (or Ig Isotype control staining) was 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 FACSymphony™ A5 Cell Analyzer System and FlowJo™ Software.
Product Details
BD Horizon™
Forkhead box A2, HNF3B, HNF-3B, HNF-3β, hepatocyte nuclear factor 3β
Human (QC Testing)
Mouse BALB/c IgG1, κ
Human FoxA2 Peptide
Intracellular staining (flow cytometry) (Routinely Tested)
5 µl/test
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
- Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
- When using high concentrations of antibody, background binding of this dye to erythroid fragments produced by ammonium chloride-based lysis, such as with BD Pharm Lyse™ Lysing Buffer (Cat. No. 555899), has been observed when the antibody conjugate was present during the lysis procedure. This may cause nonspecific staining of target cells, such as leukocytes, which have bound the resulting erythroid fragments. This background can be mitigated by any of the following: titrating the antibody conjugate to a lower concentration, fixing samples with formaldehyde, or removing erythrocytes before staining (eg, gradient centrifugation or pre-lysis with wash). This background has not been observed when cells were lysed with BD FACS™ Lysing Solution (Cat. No. 349202) after staining.
- 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.
- 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).
- For fluorochrome spectra and suitable instrument settings, please refer to our Multicolor Flow Cytometry web page at www.bdbiosciences.com/colors.
- An isotype control should be used at the same concentration as the antibody of interest.
- Please refer to http://regdocs.bd.com to access safety data sheets (SDS).
- Human donor specific background has been observed in relation to the presence of anti-polyethylene glycol (PEG) antibodies, developed as a result of certain vaccines containing PEG, including some COVID-19 vaccines. We recommend use of BD Horizon Brilliant™ Stain Buffer in your experiments to help mitigate potential background. For more information visit https://www.bdbiosciences.com/en-us/support/product-notices.
- Please observe the following precautions: We recommend that special precautions be taken (such as wrapping vials, tubes, or racks in aluminum foil) to protect exposure of conjugated reagents, including cells stained with those reagents, to any room illumination. Absorption of visible light can significantly affect the emission spectra and quantum yield of tandem fluorochrome conjugates.
- Tandem fluorochromes contain both an energy donor and an energy acceptor. Although every effort is made to minimize the lot-to-lot variation in the efficiency of the fluorochrome energy transfer, differences in the residual emission from the donor may be observed. Additionally, multi-laser cytometers may directly excite both the donor and acceptor fluorochromes. Therefore, we recommend for every tandem conjugate, a matched individual single-stain control be acquired for generating a compensation or spectral unmixing matrix.
- Cy is a trademark of Global Life Sciences Solutions Germany GmbH or an affiliate doing business as Cytiva.
- For U.S. patents that may apply, see bd.com/patents.
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
RB780 Mouse IgG1, κ Isotype Control RUO
Size
50 µg
Cat No.
568532
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571080 Rev. 1
Antibody Details
N17-280
FoxA2, forkhead box A2, is a member of the forkhead class of DNA-binding proteins that regulates gene expression in the liver, pancreatic islets, adipocytes and some neural cells. This hepatocyte nuclear factor is a transcriptional activator for liver-specific genes such as alpha fetoprotein, albumin, tyrosine aminotransferase and transthyretin. FoxA2 is expressed in embryonic endoderm, the germ layer that gives rise to the digestive system, and contributes to the specification of the pancreas and the regulation of glucose homoeostasis. FoxA2 also has roles in neural development. Specifically, FoxA2 cooperates with related FoxA1 in the specification and differentiation of midbrain dopaminergic neurons in a dosage-dependent manner.
571080 Rev. 1
Format Details
RB780
The BD Horizon RealBlue™ 780 (RB780) Dye is part of the BD family of blue dyes. It is a tandem fluorochrome with an excitation maximum (Ex Max) at 498-nm and an emission maximum (Em Max) at 781-nm. Driven by BD innovation, RB780 can be used on both spectral and conventional cytometers and is designed to be excited by the Blue laser (488-nm) with minimal excitation by the 561-nm Yellow-Green laser. For conventional instruments equipped with a Blue laser (488-nm), RB780 can be used as an alternative to PE-Cy7 and we recommend using an optical filter centered near 780-nm (eg, a 780/60-nm bandpass filter). For spectral instruments equipped with a Blue laser (488-nm), it can be used in conjunction with PE-Cy7. RB780 is on average brighter than PE-Cy7 and has minimal spillover into Yellow-Green detectors.
RB780
Blue 488 nm
498 nm
781 nm
571080 Rev.1
Citations & References
View product citations for antibody "571080" on CiteAb
Development References (7)
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Burtscher I, Lickert H.. Foxa2 regulates polarity and epithelialization in the endoderm germ layer of the mouse embryo. Development. 2009; 136(6):1029-1038. (Biology). View Reference
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Clevidence DE, Overdier DG, Tao W, et al. Identification of nine tissue-specific transcription factors of the hepatocyte nuclear factor 3/forkhead DNA-binding-domain family. Proc Natl Acad Sci U S A. 1993; 90(9):3948-3952. (Biology). View Reference
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D'Amour KA, Agulnick AD, Eliazer S, Kelly OG, Kroon E, Baetge EE. Efficient differentiation of human embryonic stem cells to definitive endoderm.. Nat Biotechnol. 2005; 23(12):1534-41. (Methodology). View Reference
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Lai E, Prezioso VR, Tao WF, Chen WS, Darnell JE Jr. Hepatocyte nuclear factor 3 alpha belongs to a gene family in mammals that is homologous to the Drosophila homeotic gene fork head. Genes Dev. 1991; 5(3):416-427. (Biology). View Reference
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Lin W, Metzakopian E, Mavromatakis YE, et al. Foxa1 and Foxa2 function both upstream of and cooperatively with Lmx1a and Lmx1b in a feedforward loop promoting mesodiencephalic dopaminergic neuron development.. Dev Biol. 2009; 333(2):386-396. (Biology). View Reference
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Monaghan AP, Kaestner KH, Grau E, Schütz G. Postimplantation expression patterns indicate a role for the mouse forkhead/HNF-3 alpha, beta and gamma genes in determination of the definitive endoderm, chordamesoderm and neuroectoderm.. Development. 1993; 119(3):567-578. (Biology). View Reference
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Thomson JA, Itskovitz-Eldor J, Shapiro SS, et al. Embryonic stem cell lines derived from human blastocysts. Science. 1998; 282:1145-1147. (Methodology). View Reference
571080 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.