Flow cytometric analysis of HIF-1α expression in human HT-1080 cells. Cells from the human HT-1080 (Fibrosarcoma, ATCC CCL-121) cell line were either treated overnight with 200 μM of Cobalt Chloride (CoCl2; Right Plot) or left untreated (DMSO only; Left Plot). The cells were fixed and permeabilized with the BD Pharmingen™ Transcription Factor Buffer Set (Cat. No. 562574/562725) and stained with either Alexa Fluor® 647 Mouse IgG1 κ Isotype Control (Cat. No. 557714; dashed line histogram) or Alexa Fluor 647 Mouse Anti-Human HIF-1α antibody (Cat. No. 565924; solid line histogram). The fluorescence histogram showing HIF-1α expression (or Ig Isotype control staining) was derived from gated events with the forward and side light-scatter characteristics of intact cells. Flow cytometric analysis was performed using a BD LSRFortessa™ Flow Cytometry System.
Flow cytometric analysis of HIF-1α expression in human HT-1080 cells. Cells from the human HT-1080 (Fibrosarcoma, ATCC CCL-121) cell line were either treated overnight with 200 μM of Cobalt Chloride (CoCl2; Right Plot) or left untreated (DMSO only; Left Plot). The cells were fixed and permeabilized with the BD Pharmingen™ Transcription Factor Buffer Set (Cat. No. 562574/562725) and stained with either Alexa Fluor® 647 Mouse IgG1 κ Isotype Control (Cat. No. 557714; dashed line histogram) or Alexa Fluor 647 Mouse Anti-Human HIF-1α antibody (Cat. No. 565924; solid line histogram). The fluorescence histogram showing HIF-1α expression (or Ig Isotype control staining) was derived from gated events with the forward and side light-scatter characteristics of intact cells. Flow cytometric analysis was performed using a BD LSRFortessa™ Flow Cytometry System.
Product Details
BD Pharmingen™
HIF1; HIF-1A; HIF1A; HIF-1-alpha; HIF1-alpha; BHLHE78; MOP1; PASD8
Human (QC Testing)
Mouse IgG1, κ
Human HIF-1α aa. 610-727
Intracellular staining (flow cytometry) (Routinely Tested)
0.2 mg/ml
AB_2739388
Aqueous buffered solution containing ≤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® 647 under optimum conditions, and unreacted Alexa Fluor® 647 was removed.
Product Notices
- Since applications vary, each investigator should titrate the reagent to obtain optimal results.
- 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® 647 fluorochrome emission is collected at the same instrument settings as for allophycocyanin (APC).
- Alexa Fluor® is a registered trademark of Molecular Probes, Inc., Eugene, OR.
- 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.
- An isotype control should be used at the same concentration as the antibody of interest.
Companion Products
Stain Buffer (FBS) RUO
Size
500 mL
Cat No.
554656
Stain Buffer (BSA) RUO
Size
500 mL
Cat No.
554657
Transcription Factor Buffer Set RUO
Size
100 Tests
Cat No.
562574
Alexa Fluor® 647 Mouse IgG1 κ Isotype Control RUO
Size
100 Tests
Cat No.
557714
Transcription Factor Buffer Set RUO
Size
25 Tests
Cat No.
562725
565924 Rev. 1
Antibody Details
54/HIF-1α
The 54/HIF-1α monoclonal antibody specifically recognizes the alpha subunit (HIF-1alpha/HIF-1a/HIF1A) of the heterodimeric helix-loop-helix transcription factor, Hypoxia-induced factor 1 (HIF-1) complex which is involved in cellular O2 homeostasis. Under hypoxic conditions, HIF-1a accumulates and translocates into the nucleus, where it binds to the constitutively-expressed HIF-1 beta subunit (HIF-1b, also known as, Aryl hydrocarbon receptor nuclear translocator/ARNT). This complex can then bind to hypoxic response elements (HREs) of target genes, including glucose transporters, glycolytic enzymes, erythropoietin, vascular endothelial growth factor (VEGF), and various other genes that facilitate metabolic adaptation to hypoxia. HIF-1a is also known to be regulated by proinflammatory cytokines and bacterial products and plays an essential role in inflammation, immunity, and cancer. HIF-1a is rapidly degraded under normoxic conditions.
565924 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 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
565924 Rev.1
Citations & References
Development References (7)
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Effects of cellular iron deficiency on the formation of vascular endothelial growth factor and angiogenesis. Iron deficiency and angiogenesis. Cancer Cell. 10(28)(Biology). View Reference
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Fallone F, Britton S, Nieto L, Salles B, Muller C. ATR controls cellular adaptation to hypoxia through positive regulation of hypoxia-inducible factor 1 (HIF-1) expression. Oncogene. 2013; 32(37):4387-4396. (Biology). View Reference
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Lee JW, Bae SH, Jeong JW, Kim SH, Kim KW. Hypoxia-inducible factor (HIF-1)alpha: its protein stability and biological functions.. Exp Mol Med. 2004; 36(1):1-12. (Biology). View Reference
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Poitz DM, Augstein A, Weinert S, Braun-Dullaeus RC, Strasser RH, Schmeisser A. OxLDL and macrophage survival: essential and oxygen-independent involvement of the Hif-pathway.. Basic Res Cardiol. 2011; 106(5):761-72. (Biology). View Reference
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Raval RR, Lau KW, Tran MG, et al. Contrasting properties of hypoxia-inducible factor 1 (HIF-1) and HIF-2 in von Hippel-Lindau-associated renal cell carcinoma.. Mol Cell Biol. 2005; 25(13):5675-86. (Clone-specific: Immunohistochemistry, Western blot). View Reference
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Zagzag D, Lukyanov Y, Lan L, et al. Hypoxia-inducible factor 1 and VEGF upregulate CXCR4 in glioblastoma: implications for angiogenesis and glioma cell invasion.. Lab Invest. 2006; 86(12):1221-32. (Clone-specific: Immunofluorescence, Immunohistochemistry, Western blot). View Reference
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van der Groep P, Bouter A, Menko FH, van der Wall E, van Diest PJ. High frequency of HIF-1alpha overexpression in BRCA1 related breast cancer.. Breast Cancer Res Treat. 2008; 111(3):475-80. (Clone-specific: Immunohistochemistry). View Reference
565924 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.
