Multicolor flow cytometric analysis of Hobit (ZNF683) expression in human peripheral blood lymphocytes. Human peripheral blood mononuclear cells were fixed and permeabilized using the BD Pharmingen™ Transcription Factor Buffer Set (Cat. No. 562574/562725). The cells were then stained with BD Horizon™ BV421 Mouse Anti-Human CD3 (Cat. No. 562426/562427), PE Mouse Anti-Human CD56 (Cat. No. 563238), and Alexa Fluor® 647 Mouse Anti-Human Hobit (ZNF683) (Cat. No. 566250) antibodies. Two-color flow cytometric contour plots showing the correlated expression of CD3 versus Hobit (ZNF683) (Left Plot) or CD56 versus Hobit (ZNF683) (Right Plot) were derived from gated events with the forward and side light-scatter characteristics of intact lymphocytes. Flow cytometric analysis was performed using a BD LSRFortessa™ Cell Analyzer System.
Multicolor flow cytometric analysis of Hobit (ZNF683) expression in human peripheral blood lymphocytes. Human peripheral blood mononuclear cells were fixed and permeabilized using the BD Pharmingen™ Transcription Factor Buffer Set (Cat. No. 562574/562725). The cells were then stained with BD Horizon™ BV421 Mouse Anti-Human CD3 (Cat. No. 562426/562427), PE Mouse Anti-Human CD56 (Cat. No. 563238), and Alexa Fluor® 647 Mouse Anti-Human Hobit (ZNF683) (Cat. No. 566250) antibodies. Two-color flow cytometric contour plots showing the correlated expression of CD3 versus Hobit (ZNF683) (Left Plot) or CD56 versus Hobit (ZNF683) (Right Plot) were derived from gated events with the forward and side light-scatter characteristics of intact lymphocytes. Flow cytometric analysis was performed using a BD LSRFortessa™ Cell Analyzer System.
Product Details
BD Pharmingen™
HOBIT; Homolog of Blimp-1 in T cells; Zinc finger protein 683; ZNF683
Human (QC Testing)
Mouse BALB/c IgM, κ
Human Hobit Recombinant Protein
Intracellular staining (flow cytometry) (Routinely Tested)
5 µl
AB_2739629
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® 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).
- 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.
- 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.
- 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
BV421 Mouse Anti-Human CD3 RUO
Size
100 Tests
Cat No.
562426
BV421 Mouse Anti-Human CD3 RUO
Size
25 Tests
Cat No.
562427
PE Mouse Anti-Human NCAM-1 (CD56) RUO
Size
50 Tests
Cat No.
563238
566250 Rev. 1
Antibody Details
Sanquin-Hobit/1
The Sanquin-Hobit/1 monoclonal antibody specifically binds to Homolog of Blimp-1 in T cells (Hobit), which is also known as Zinc finger protein 683 (ZNF683). Hobit is a conserved Zn-finger transcription factor that is homologous to Blimp-1. It is expressed in cytotoxic effector-type CD8+ T cells and in some effector memory CD8+ T cells, but is not significantly expressed in naïve or most central memory CD8+ T cells. Hobit is also expressed in natural killer (NK) cells, and NKT cells, but is not expressed in CD4+ T cells, B cells, monocytes, dendritic cells, or neutrophils. Hobit expression is strongly correlated with the expression T-bet and Interferon-γ (IFN-γ) mRNA. Hobit appears to play a role in the differentiation and effector functions of NK cells and effector CD8+ T cells such as in the production of IFN-γ.
566250 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
566250 Rev.1
Citations & References
Development References (3)
-
Braun J, Frentsch M, Thiel A. Hobit and human effector T-cell differentiation: The beginning of a long journey.. Eur J Immunol. 2015; 45(10):2762-5. (Biology). View Reference
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Vieira Braga FA, Hertoghs KM, Kragten NA, et al. Blimp-1 homolog Hobit identifies effector-type lymphocytes in humans.. Eur J Immunol. 2015; 45(10):2945-58. (Immunogen: Flow cytometry). View Reference
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van Gisbergen KP, Kragten NA, Hertoghs KM, et al. Mouse Hobit is a homolog of the transcriptional repressor Blimp-1 that regulates NKT cell effector differentiation.. Nat Immunol. 2012; 13(9):864-71. (Biology). View Reference
566250 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.
