Two-parameter flow cytometric analysis of HuR expression in human peripheral blood mononuclear cell populations. Human PBMCs were fixed and permeabilized using the BD Pharmingen™ Transcription Factor Buffer Set (Cat. No. 562574/562725) followed by staining with either PE Mouse IgG1, κ Isotype Control (Cat. No. 559320; Left Plot) or PE Mouse Anti-HuR antibody (Cat. No. 566341; Right Plot) at 0.063 µg/test. Two-parameter flow cytometric contour plots showing the correlated expression of HuR (or Ig Isotype control staining) versus side light-scatter (SSC-A) signals were derived from gated events with the forward and side light-scattering characteristics of intact mononuclear leucocyte populations. Flow cytometric analysis was performed using a BD LSRFortessa™ Cell Analyzer System X-20. Data shown on this Technical Data Sheet are not lot specific.
Flow cytometric analysis of HuR expression in mouse spleen cells. C57BL/6 mouse splenic leucocytes were fixed and permeabilized using the BD Pharmingen™ Transcription Factor Buffer Set followed by staining with either PE Mouse IgG1, κ Isotype Control (dashed line histogram) or PE Mouse Anti-HuR antibody (solid line histogram) at 0.063 µg/test. The fluorescence histograms were derived from the gated events with the forward and side light-scattering characteristics of intact leucocytes. Flow cytometric analysis was performed using a BD LSRFortessa™ Cell Analyzer System. Data shown on this Technical Data Sheet are not lot specific.
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Two-parameter flow cytometric analysis of HuR expression in human peripheral blood mononuclear cell populations. Human PBMCs were fixed and permeabilized using the BD Pharmingen™ Transcription Factor Buffer Set (Cat. No. 562574/562725) followed by staining with either PE Mouse IgG1, κ Isotype Control (Cat. No. 559320; Left Plot) or PE Mouse Anti-HuR antibody (Cat. No. 566341; Right Plot) at 0.063 µg/test. Two-parameter flow cytometric contour plots showing the correlated expression of HuR (or Ig Isotype control staining) versus side light-scatter (SSC-A) signals were derived from gated events with the forward and side light-scattering characteristics of intact mononuclear leucocyte populations. Flow cytometric analysis was performed using a BD LSRFortessa™ Cell Analyzer System X-20. Data shown on this Technical Data Sheet are not lot specific.
Flow cytometric analysis of HuR expression in mouse spleen cells. C57BL/6 mouse splenic leucocytes were fixed and permeabilized using the BD Pharmingen™ Transcription Factor Buffer Set followed by staining with either PE Mouse IgG1, κ Isotype Control (dashed line histogram) or PE Mouse Anti-HuR antibody (solid line histogram) at 0.063 µg/test. The fluorescence histograms were derived from the gated events with the forward and side light-scattering characteristics of intact leucocytes. Flow cytometric analysis was performed using a BD LSRFortessa™ Cell Analyzer System. Data shown on this Technical Data Sheet are not lot specific.
Product Details
BD Pharmingen™
HUR; ELAVL1; ELAV1; Hua; MelG
Mouse (QC Testing), Human (Tested in Development)
Mouse BALB/c IgG1, κ
Human HuR Recombinant Protein
Intracellular staining (flow cytometry) (Routinely Tested)
0.2 mg/ml
AB_2739698
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 with R-PE under optimum conditions, and unconjugated antibody and free PE were removed.
Product Notices
- Since applications vary, each investigator should titrate the reagent to obtain optimal results.
- 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.
- 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
Transcription Factor Buffer Set RUO
Size
100 Tests
Cat No.
562574
PE Mouse IgG1, κ Isotype Control RUO
Size
100 Tests
Cat No.
559320
Transcription Factor Buffer Set RUO
Size
25 Tests
Cat No.
562725
566341 Rev. 1
Antibody Details
3A2
The 3A2 monoclonal antibody specifically recognizes HuR (Human antigen R) which is encoded by ELAVL1 (ELAV like RNA binding protein 1). HuR belongs to the ELAVL (embryonic lethal, abnormal vision and Drosophila-like) family of proteins that includes HuB (ELAVL2), HuC (ELAVL3) and HuD (ELAVL4). These proteins contain three RNA recognition motifs that selectively bind AU rich elements (AREs) found in the 3' untranslated regions of messenger RNAs. Although predominantly located in the nucleus, HuR shuttles between the nucleus and cytoplasm. HuR stabilizes ARE-containing mRNAs, regulates RNA splicing, and can thus influence the outcome of gene expression. HuR is expressed by a variety of cell types including leucocytes, eg, T cells, B cells, and dendritic cells, whereas the other ELAVL family members are primarily expressed by neurons. HuR is involved in regulating cellular growth and differentiation, eg, through the stabilization and shuttling of mRNAs including those encoding transcription factors, proto-oncogenes, costimulatory receptors, growth factors, cytokines, chemokines, and inflammatory mediators. Abnormal expression of HuR has been linked to a number of diseases, for example, overexpressed HuR levels have been detected in certain tumor cells. The 3A2 antibody crossreacts with mouse HuR as well as with human HuB and HuD, but not with HuC.
566341 Rev. 1
Format Details
PE
R-Phycoerythrin (PE), is part of the BD family of Phycobiliprotein dyes. This fluorochrome is a multimeric fluorescent phycobiliprotein with excitation maximum (Ex Max) of 496 nm and 566 nm and an emission maximum (Em Max) at 576 nm. PE is designed to be excited by the Blue (488 nm), Green (532 nm) and Yellow-Green (561 nm) lasers and detected using an optical filter centered near 575 nm (e.g., a 575/26-nm bandpass filter). As PE is excited by multiple lasers, this can result in cross-laser excitation and fluorescence spillover on instruments with various combinations of Blue, Green, and Yellow-Green lasers. Please ensure that your instrument’s configurations (lasers and optical filters) are appropriate for this dye.
PE
Yellow-Green 488 nm, 532 nm, 561 nm
496 nm, 566 nm
576 nm
566341 Rev.1
Citations & References
Development References (11)
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Casolaro V, Fang X, Tancowny B, et al. Posttranscriptional regulation of IL-13 in T cells: role of the RNA-binding protein HuR.. J Allergy Clin Immunol. 2008; 121(4):853-9.e4. (Clone-specific: Western blot). View Reference
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Diaz-Muñoz MD, Bell SE, Fairfax K, et al. The RNA-binding protein HuR is essential for the B cell antibody response.. Nat Immunol. 2015; 16(4):415-25. (Clone-specific: Flow cytometry, Immunofluorescence, Western blot). View Reference
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Fan XC, Steitz JA. Overexpression of HuR, a nuclear-cytoplasmic shuttling protein, increases the in vivo stability of ARE-containing mRNAs.. EMBO J. 1998; 17(12):3448-60. (Biology). View Reference
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Fries B, Heukeshoven J, Hauber I, et al. Analysis of nucleocytoplasmic trafficking of the HuR ligand APRIL and its influence on CD83 expression.. J Biol Chem. 2007; 282(7):4504-15. (Biology). View Reference
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Gallouzi IE, Brennan CM, Stenberg MG, et al. HuR binding to cytoplasmic mRNA is perturbed by heat shock.. Proc Natl Acad Sci USA. 2000; 97(7):3073-8. (Immunogen: ELISA, Fluorescence microscopy, Immunofluorescence, Immunoprecipitation, Western blot). View Reference
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Gubin MM, Techasintana P, Magee JD, et al. Conditional knockout of the RNA-binding protein HuR in CD4⁺ T cells reveals a gene dosage effect on cytokine production.. Mol Med. 2014; 20:93-108. (Clone-specific: Flow cytometry). View Reference
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Ishimaru D, Ramalingam S, Sengupta TK, et al. Regulation of Bcl-2 expression by HuR in HL60 leukemia cells and A431 carcinoma cells.. Mol Cancer Res. 2009; 7(8):1354-66. (Biology). View Reference
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Mou Z, You J, Xiao Q, et al. HuR posttranscriptionally regulates early growth response-1 (Egr-1) expression at the early stage of T cell activation.. FEBS Lett. 2012; 586(24):4319-25. (Biology). View Reference
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Stellato C, Gubin MM, Magee JD, et al. Coordinate regulation of GATA-3 and Th2 cytokine gene expression by the RNA-binding protein HuR.. J Immunol. 2011; 187(1):441-9. (Clone-specific: Flow cytometry, Immunoprecipitation, Western blot). View Reference
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Techasintana P, Davis JW, Gubin MM, Magee JD, Atasoy U. Transcriptomic-Wide Discovery of Direct and Indirect HuR RNA Targets in Activated CD4+ T Cells.. PLoS ONE. 2015; 10(7):e0129321. (Clone-specific: Immunoprecipitation). View Reference
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Wang JG, Collinge M, Ramgolam V, et al. LFA-1-dependent HuR nuclear export and cytokine mRNA stabilization in T cell activation.. J Immunol. 2006; 176(4):2105-13. (Biology). View Reference
566341 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.
