BD Pharmingen™ Alexa Fluor® 488 Mouse Anti-Human Alpha-fetoprotein
Clone C3/AFP (also known as C3)
(RUO)
Flow cytometric analysis of AFP expression in human liver hepatocellular carcinoma cells. Human liver hepatocellular carcinoma cells Hep G2 (ATCC®, HB-8065™) were harvested with Accutase™ Cell Detachment Solution (Cat. No. 561527), 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 Alexa Fluor® 488 Mouse IgG2a, κ isotype control (dashed line, Cat. No. 558055) or Alexa Fluor® 488 Mouse anti-Human Alpha-fetoprotein monoclonal antibody (solid line) at matched concentrations. Histograms were derived from gated events based on light scattering characteristics of HepG2 cells. Flow cytometry was performed on a BD FACSCanto™ II flow cytometry system.
Image analysis of AFP expressed in human hepatocellular carcinoma cell line. Hep G2 cells (ATCC®; HB-8065™) were fixed with BD Cytofix™ Buffer (Cat. No. 554655), permeabilized with 0.1% Triton™ X-100 (Sigma), and stained with Alexa Fluor® 488 Mouse Anti-Human Alpha-fetoprotein monoclonal antibody (pseudo-colored green). Counter-staining was with Hoechst (pseudo-colored blue). The image was captured using a BD Pathway™ 435 Cell Analyzer and merged using BD Attovision™ Software.
.png){kind=spacer}
Flow cytometric analysis of AFP expression in human liver hepatocellular carcinoma cells. Human liver hepatocellular carcinoma cells Hep G2 (ATCC®, HB-8065™) were harvested with Accutase™ Cell Detachment Solution (Cat. No. 561527), 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 Alexa Fluor® 488 Mouse IgG2a, κ isotype control (dashed line, Cat. No. 558055) or Alexa Fluor® 488 Mouse anti-Human Alpha-fetoprotein monoclonal antibody (solid line) at matched concentrations. Histograms were derived from gated events based on light scattering characteristics of HepG2 cells. Flow cytometry was performed on a BD FACSCanto™ II flow cytometry system.
Image analysis of AFP expressed in human hepatocellular carcinoma cell line. Hep G2 cells (ATCC®; HB-8065™) were fixed with BD Cytofix™ Buffer (Cat. No. 554655), permeabilized with 0.1% Triton™ X-100 (Sigma), and stained with Alexa Fluor® 488 Mouse Anti-Human Alpha-fetoprotein monoclonal antibody (pseudo-colored green). Counter-staining was with Hoechst (pseudo-colored blue). The image was captured using a BD Pathway™ 435 Cell Analyzer and merged using BD Attovision™ Software.
Product Details
BD Pharmingen™
Alpha-1-fetoprotein, Alpha-fetoglobulin, AFP
Human (QC Testing)
Mouse BALB/c IgG2a
Human Alpha-fetoprotein
Intracellular staining (flow cytometry) (Routinely Tested), Bioimaging, Immunofluorescence (Tested During Development)
5 µl
AB_2737952
Aqueous buffered solution containing BSA, protein stabilizer, 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 to Alexa Fluor® 488 under optimum conditions, and unreacted Alexa Fluor® 488 was removed.
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.
- Alexa Fluor® 488 fluorochrome emission is collected at the same instrument settings as for fluorescein isothiocyanate (FITC).
- 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.
- Accutase is a registered trademark of Innovative Cell Technologies, Inc.
- Triton is a trademark of the Dow Chemical Company.
- All other brands are trademarks of their respective owners.
- 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.
- For fluorochrome spectra and suitable instrument settings, please refer to our Multicolor Flow Cytometry web page at www.bdbiosciences.com/colors.
- Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
Companion Products
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
Alexa Fluor®488 Mouse IgG2a, κ Isotype control RUO
Size
50 Tests
Cat No.
558055
.png?imwidth=320)
Accutase™ Cell Detachment Solution RUO
Size
100 mL
Cat No.
561527
563016 Rev. 1
Antibody Details
C3/AFP
Alpha-fetoprotein (AFP) is a member of the albuminoid gene family which includes albumin, alpha-albumin, and vitamin D protein. Alpha-fetoprotein is an abundant plasma protein synthesized from fetal yolk sac, liver, and gastrointestinal tract during development. Similar to albumin, AFP binds and transports multiple ligands such as nickel, copper, billirubin, and fatty acids. AFP levels are low to absent in healthy adult tissues whereas elevated AFP levels in the adult can be indicative of malignancies. In particular, AFP is expressed in hepatocellular carcinoma, germ cell tumors, and metatstatic cancers of the liver. Maternal serum AFP levels can be monitored to screen for fetal abnormalities. AFP is used as a hepatic progenitor marker in hepatic differentiations from pluripotent stem cells.
563016 Rev. 1
Format Details
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
563016 Rev.1
Citations & References
View product citations for antibody "563016" on CiteAb
Development References (5)
-
Duan Y, Catana A, Meng Y, et al. Differentiation and enrichment of hepatocyte-like cells from human embryonic stem cells in vitro and in vivo. Stem Cells. 2007; 25(15):3058-3068. (Biology). View Reference
-
Jozefczuk J, Prigione A, Chavez L, Adjaye J. Comparative analysis of human embryonic stem cell and induced pluripotent stem cell-derived hepatocyte-like cells reveals current drawbacks and possible strategies for improved differentiation. Stem Cells Dev. 2011; 20(7):1259-1275. (Biology). View Reference
-
Mizejewski GJ. Alpha-fetoprotein structure and function: relevance to isoforms, epitopes, and conformational variants. Exp Biol Med (Maywood). 2001; 226(5):377-408. (Biology). View Reference
-
Yazova AK, Goussev AI, Christiansen M, et al. Human fetal and tumor alpha-fetoproteins differ in conformationally dependent epitope variants expression. Immunol Lett. 2003; 85(3):261-270. (Clone-specific). View Reference
-
Yazova AK, Goussev AI, Poltoranina VS, Yakimenko EF. Human alpha-fetoprotein epitopes as revealed by monoclonal antibodies. Immunol Lett. 1990; 25(4):325-330. (Clone-specific). View Reference
563016 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.