Flow cytometric analysis of Heme oxygenase 1 expression in human peripheral blood mononuclear cells. Peripheral blood mononuclear cells (PBMCs) were fixed and permeabilized with BD Cytofix/Cytoperm™ Fixation and Permeabilization Solution (Cat. No. 554722). The cells were then washed and stained in BD Perm/Wash™ Buffer (Cat. No. 554723) with BD Horizon™ BV421 Mouse Anti-Human CD14 antibody (Cat. No. 565283) and either Alexa Fluor® 647 Mouse IgG1, κ Isotype Control (Cat. No.557732; Left Plot) or Alexa Fluor® 647 Mouse Anti-Human Heme oxygenase 1 antibody (Cat. No. 566391; Right Plot). Two-color flow cytometric contour plots showing the correlated expression of Heme oxygenase 1 (or Ig Isotype control staining) versus CD14 were derived from gated events with the forward and side light-scatter characteristics of intact leucocytes. Flow cytometric analysis was performed using a BD LSRFortessa™ X-20 Cell Analyzer System. Data shown on this Technical Data Sheet are not lot specific.
Flow cytometric analysis of Heme oxygenase 1 expression in human peripheral blood mononuclear cells. Peripheral blood mononuclear cells (PBMCs) were fixed and permeabilized with BD Cytofix/Cytoperm™ Fixation and Permeabilization Solution (Cat. No. 554722). The cells were then washed and stained in BD Perm/Wash™ Buffer (Cat. No. 554723) with BD Horizon™ BV421 Mouse Anti-Human CD14 antibody (Cat. No. 565283) and either Alexa Fluor® 647 Mouse IgG1, κ Isotype Control (Cat. No.557732; Left Plot) or Alexa Fluor® 647 Mouse Anti-Human Heme oxygenase 1 antibody (Cat. No. 566391; Right Plot). Two-color flow cytometric contour plots showing the correlated expression of Heme oxygenase 1 (or Ig Isotype control staining) versus CD14 were derived from gated events with the forward and side light-scatter characteristics of intact leucocytes. Flow cytometric analysis was performed using a BD LSRFortessa™ X-20 Cell Analyzer System. Data shown on this Technical Data Sheet are not lot specific.
Product Details
BD Pharmingen™
HMOX1; HO-1; Heat shock protein 32-kD; HSP32
Human (QC Testing)
Mouse IgG1
Human Heme Oxygenase 1 (HO-1) aa. 150-286
Flow cytometry (Routinely Tested)
5 µl
AB_2739722
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 to Alexa Fluor® 647 under optimum conditions, and unreacted Alexa Fluor® 647 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.
- 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.
- Source of all serum proteins is from USDA inspected abattoirs located in the United States.
- 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.
- Alexa Fluor® 647 fluorochrome emission is collected at the same instrument settings as for allophycocyanin (APC).
- 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
Stain Buffer (BSA) RUO
Size
500 mL
Cat No.
554657
Fixation and Permeabilization Solution RUO
Size
125 mL
Cat No.
554722
Perm/Wash Buffer RUO
Size
100 mL
Cat No.
554723
Alexa Fluor® 647 Mouse IgG1 κ Isotype Control RUO
Size
100 Tests
Cat No.
557732
BV421 Mouse Anti-Human CD14 RUO
Size
50 Tests
Cat No.
565283
566391 Rev. 1
Antibody Details
23/Heme Oxygenase 1
The 23/Heme Oxygenase 1 monoclonal antibody specifically recognizes Heme oxygenase 1 (HO-1), also known as Heat Shock Protein 32 (HSP32). HO-1 is a 32 kDa monooxygenase encoded by HMOX1. Heme oxygenases 1 and 2 (HO-1, HO-2) cleave the heme molecule, resulting in the production of carbon monoxide (CO) and biliverdin. While HO-2 is constitutively expressed in tissues, HO-1 is rapidly induced by several stimuli such as lutathione depletion, hemin, heat shock, heavy metals, oxidative stress, oxidized LDL, anoxia, and endotoxic shock. HO-1 and its byproducts have potent antioxidant, antiproliferative, anti-inflammatory and anti-apoptotic effects in various cell types. In humans, HO-1 deficiency is associated with susceptibility to oxidative stress, increased inflammation and endothelial cell damage. In immune cells, HO-1 is required for the activation of interferon (IFN) regulatory factor 3 (IRF3) after Toll-like receptor 3 or 4 stimulation or viral infection. It is also upregulated on T cells upon TCR stimulation.
566391 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
566391 Rev.1
Citations & References
View product citations for antibody "566391" on CiteAb
Development References (8)
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Foresti R, Clark JE, Green CJ, Motterlini R. Thiol compounds interact with nitric oxide in regulating heme oxygenase-1 induction in endothelial cells. Involvement of superoxide and peroxynitrite anions. J Biol Chem. 1997; 272(29):18411-18417. (Biology). View Reference
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Fürst R, Blumenthal SB, Kiemer AK, Zahler S, Vollmar AM. Nuclear factor-kappa B-independent anti-inflammatory action of salicylate in human endothelial cells: induction of heme oxygenase-1 by the c-jun N-terminal kinase/activator protein-1 pathway.. J Pharmacol Exp Ther. 2006; 318(1):389-94. (Clone-specific: Western blot). View Reference
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Ishikawa K, Navab M, Leitinger N, Fogelman AM, Lusis AJ. Induction of heme oxygenase-1 inhibits the monocyte transmigration induced by mildly oxidized LDL. J Clin Invest. 1997; 100(5):1209-1216. (Biology). View Reference
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McAllister SC, Hansen SG, Ruhl RA, et al. Kaposi sarcoma-associated herpesvirus (KSHV) induces heme oxygenase-1 expression and activity in KSHV-infected endothelial cells.. Blood. 2004; 103(9):3465-73. (Clone-specific: Immunofluorescence). View Reference
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Suzuki M, Ishizaka N, Tsukamoto K. Pressurization facilitates adenovirus-mediated gene transfer into vein graft. FEBS Lett. 2000; 470(3):370-374. (Clone-specific: Western blot). View Reference
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Wang X, Mazurkiewicz M, Hillert EK, et al. The proteasome deubiquitinase inhibitor VLX1570 shows selectivity for ubiquitin-specific protease-14 and induces apoptosis of multiple myeloma cells.. Sci Rep. 2016; 6:26979. (Clone-specific: Western blot). View Reference
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Yoshida T, Biro P, Cohen T, Muller RM, Shibahara S. Human heme oxygenase cDNA and induction of its mRNA by hemin. Eur J Biochem. 1988; 171(3):457-461. (Biology). View Reference
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Zhong H, Bao W, Friedman D, Yazdanbakhsh K. Hemin controls T cell polarization in sickle cell alloimmunization.. J Immunol. 2014; 193(1):102-10. (Clone-specific: Flow cytometry). View Reference
566391 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.