Skip to main content Skip to navigation
Hamburger Menu
Search icon
Country Selector Country Selector
Spain (English)
Profile Icon Profile Icon
Sign-in/Register UserName
Products

Solutions

Discover & Learn

Resources & Tools

Support

Search icon Search icon
Close Menu
      Alert Icon
      PE Mouse Anti-Human IP-10
      PE Mouse Anti-Human IP-10
      Expression of IP-10 by stimulated human monocytes. Human PBMC were stimulated for 24 hours with Human Interferon-γ (1500 U/ml final concentration; Cat. No. 554617) in the presence of GolgiStop™ (2 µM final concentration; Cat. No. 554724). The PBMC were harvested, fixed, permeabilized, and stained with 0.05 µg of PE-mouse anti-human IP-10 antibody (PE-6D4/D6/G2, Cat. No. 555049) following BD Pharmingen staining protocol (see Figure, left panel). The data reflect gating on monocytes, based on forward and side scattered light signals. To demonstrate specificity of staining, binding by the PE-6D4/D6/G2 antibody was blocked by preincubation of the fixed/permeabilized cells with excess unlabeled 6D4/D6/G2 antibody (5 µg; middle panel) prior to staining with the PE-6D4/D6/G2 antibody. The level of nonspecific staining was assessed using the PE-mouse IgG2a isotype control (0.05 µg; PE-G155-178; Cat. No. 554648; right panel). The quadrant markers for the bivariate dot plots were set based on the autofluorescence control and verified using the unlabeled antibody blocking control.
      PE Mouse Anti-Human IP-10
      Expression of IP-10 by stimulated human monocytes. Human PBMC were stimulated for 24 hours with Human Interferon-γ (1500 U/ml final concentration; Cat. No. 554617) in the presence of GolgiStop™ (2 µM final concentration; Cat. No. 554724). The PBMC were harvested, fixed, permeabilized, and stained with 0.05 µg of PE-mouse anti-human IP-10 antibody (PE-6D4/D6/G2, Cat. No. 555049) following BD Pharmingen staining protocol (see Figure, left panel). The data reflect gating on monocytes, based on forward and side scattered light signals. To demonstrate specificity of staining, binding by the PE-6D4/D6/G2 antibody was blocked by preincubation of the fixed/permeabilized cells with excess unlabeled 6D4/D6/G2 antibody (5 µg; middle panel) prior to staining with the PE-6D4/D6/G2 antibody. The level of nonspecific staining was assessed using the PE-mouse IgG2a isotype control (0.05 µg; PE-G155-178; Cat. No. 554648; right panel). The quadrant markers for the bivariate dot plots were set based on the autofluorescence control and verified using the unlabeled antibody blocking control.
      Product Details
      Down Arrow Up Arrow


      BD Pharmingen™
      Human (QC Testing)
      Mouse IgG2a, κ
      Recombinant human IP-10 protein
      Intracellular staining (flow cytometry) (Routinely Tested)
      0.2 mg/ml
      AB_395670
      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.
      Show More blue down arrow Show Less blue up arrow

      Recommended Assay Procedures

      Immunofluorescent Staining and Flow Cytometric Analysis: The PE-conjugated 6D4/D6/G2 antibody (Cat. No. 555049) can be used for multicolor immunofluorescent staining and flow cytometric analyses to identify and enumerate human IP-10-producing cells within mixed cell populations (See Figure).  For optimal immunofluorescent staining with flow cytometric analysis, this anti-cytokine antibody should be titrated.  (≤ 0.5 µg mAb/1X10^6 cells) For specific methodology, please see the online protocols or the chapter on intracellular staining in the Immune Function handbook posted at www.bdbiosciences.com.

      A useful control for demonstrating specificity of staining is to pre-block the paraformaldehyde-fixed/saponin-permeabilized cells with unlabeled 6D4/D6/G2 antibody prior to staining. The staining technique and use of blocking controls are described by C. Prussin, et al. A suitable mouse IgG2a isotype control for assessing the level of background staining on paraformaldehyde-fixed/saponin-permeabilized human cells is PE-G155-178 (Cat. No. 554648); use at comparable concentrations to antibody of interest (e.g., ≤ 0.5 µg mAb/1X10^6 cells).

      ELISA: The biotinylated 6D4/D6/G2 (Cat. No. 555048) antibody is useful as a detection antibody for a sandwich ELISA for specifically measuring human IP-10 protein levels. Biotin 6D4/D6/G2 antibody can be paired with the Purified 4D5/A7/C5 antibody (Cat. No. 555046) as the capture antibody and with Recombinant human IP-10 protein (Cat. No. 551130) as the standard.

      Show More blue down arrow Show Less blue up arrow

      Product Notices

      1. Since applications vary, each investigator should titrate the reagent to obtain optimal results.
      2. For fluorochrome spectra and suitable instrument settings, please refer to our Multicolor Flow Cytometry web page at www.bdbiosciences.com/colors.
      3. 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.
      4. An isotype control should be used at the same concentration as the antibody of interest.
      5. Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
      Show More blue down arrow Show Less blue up arrow
      555049 Rev. 2
      Antibody Details
      Down Arrow Up Arrow
      6D4/D6/G2

      The monoclonal antibody 6D4/D6/G2 reacts with human CXC chemokine, interferon gamma inducible protein 10 (IP-10). IP-10 is inducible in monocytes, keratinocytes and endothelial cells by IFN-γ. The immunogen used to generate the monoclonal antibody 6D4/D6/G2 was recombinant human IP-10 protein.

      555049 Rev. 2
      Format Details
      Down Arrow Up Arrow
      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.
      altImg
      PE
      Yellow-Green 488 nm, 532 nm, 561 nm
      496 nm, 566 nm
      576 nm
      555049 Rev.2
      Citations & References
      Down Arrow Up Arrow
      View product citations for antibody "555049" on CiteAb

      Development References (3)

      1. Luster AD, Ravetch JV. Biochemical characterization of a gamma interferon-inducible cytokine (IP-10). J Exp Med. 1987; 166(4):1084-1097. (Clone-specific). View Reference
      2. Luster AD, Unkeless JC, Ravetch JV. Gamma-interferon transcriptionally regulates an early-response gene containing homology to platelet proteins. Nature. 1985; 315(6021):672-676. (Clone-specific). View Reference
      3. Prussin C, Metcalfe DD. Detection of intracytoplasmic cytokine using flow cytometry and directly conjugated anti-cytokine antibodies. J Immunol Methods. 1995; 188(1):117-128. (Methodology: IC/FCM Block). View Reference
      555049 Rev. 2

      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.