Monoclonal mouse anti human Interferon alpha

///Monoclonal mouse anti human Interferon alpha

Monoclonal mouse anti human Interferon alpha

$99.00$149.00


accession P01563


Applications

ELISA: This antibody can be used as capture antibody at 2 μg/mL jointly with biotinylated Anti IFN-a polyclonal detection antibody to detect Human IFN-alpha. The detection limit for recombinant Human IFNA is approximately 0.1 ng/well.

Western blot:This monoclonal antibody has been tested in WB analysis of human IFNA . A suitable range of concentrations of this antibody for WB detection is 1-2 µg/ml.
The detection limit for Recombinant Human Interferon alpha is 1.0-2.0 ng/lane, under reducing or non-reducing conditions.

Source This antibody was produced from a hybridoma resulting from the fusion of a mouse myeloma with B cells obtained from a mouse immunized with recombinant E. coli-derived Human IFNa.
Species reactivity Human Interferon alpha.
Purification The IgG fraction of the tissue culture supernatant was purified by Protein A affinity chromatography.
Presentation Prepared from sterile filtered solution at a concentration of 1mg/ml in PBS with 5% trehalose.
Reconstitution A quick spin of the vial followed by reconstitution in distilled water to a concentration not less than 0.1 mg/mL. This solution can then be diluted into other buffers
Storage This antibody can be stored in working aliquots at 2° - 8° C for one month, or at -20° C for six months without detectable loss of activity. Avoid repeated freeze/thaw cycles.
Usage This antibody product is for research purposes only.It may not be used for therapeutics or diagnostic purposes.


Interactor P17181
Interactor P48551 INAR2_HUMAN
Biological Process Antiviral-defense
Molecular function Cytokine

Methods

iNKT cell activation assays

  • Activation of iNKT cells after coculture with DCs from 81A, 81A-eGFP, or mock-infected cultures was assessed by flow cytometry and microscopy.
  • For the flow cytometry assay, DCs were preincubated with different concentrations of α-GalCer (0.1–100 ng/mL) overnight at 37°C before adding iNKT cells at a ratio of 1:2.
  • After 6 h of coculture in the presence of brefeldin A (2 mg/mL ), cells were surface-stained with anti-CD3 and anti-CD11c mAb to discriminate between DCs and iNKT cells.
  • Following permeabilization, cells were stained with anti-IFN-γ mAb and analyzed on a .
  • To assess iNKT cell activation by microcopy, iNKT cells were coincubated with DCs preloaded with α-GalCer (100 ng/mL) at a 1:2 ratio in the presence of brefeldin A.
  • Minimal coincubation times for complex formation and cytokine detection were 30 min for TNF-α and 2 h for IFN-γ.
  • After fixation with 4% paraformaldehyde, cell complexes were cytospun onto glass…
  • Activation of iNKT cells after coculture with DCs from 81A, 81A-eGFP, or mock-infected cultures was assessed by flow cytometry and microscopy.
  • For the flow cytometry assay, DCs were preincubated with different concentrations of α-GalCer (0.1–100 ng/mL) overnight at 37°C before adding iNKT cells at a ratio of 1:2.
  • After 6 h of coculture in the presence of brefeldin A (2 mg/mL ), cells were surface-stained with anti-CD3 and anti-CD11c mAb to discriminate between DCs and iNKT cells.
  • Following permeabilization, cells were stained with anti-IFN-γ mAb and analyzed on a .
  • To assess iNKT cell activation by microcopy, iNKT cells were coincubated with DCs preloaded with α-GalCer (100 ng/mL) at a 1:2 ratio in the presence of brefeldin A.
  • Minimal coincubation times for complex formation and cytokine detection were 30 min for TNF-α and 2 h for IFN-γ.
  • After fixation with 4% paraformaldehyde, cell complexes were cytospun onto glass microscope slides using a Cytospin 4 cytoentrifuge (800 rpm, 5 min , , ) and permeabilized with 0.1% saponin .
  • Cells were stained with anti-IFN-γ mAb (25,718; 1:50& ) or anti-TNF-α mAb (28,401; 1:50& ), blocked with 1% normal goat serum (ako, , enmark), and finally, incubated with goat anti-mouse Alexa-594 secondary antibody (1:500 , , O, ) and API .
  • All incubations with primary and secondary antibodies and serum were at room temperature for 30 min.
  • Images were obtained on a Nikon A1R confocal system with a 60×/1.49 oil objective using NIS-Elements AR software (3.2).
  • IFN-γ production by iNKT cells was analyzed by visual inspection of randomly taken images of DC-iNKT cell complexes.
  • To avoid operator bias, image acquisition and analysis were performed independently by different investigators.

Read more

MIF, VEGF and sFlt-1 expression in physiological placental villous explants treated with culture media conditioned by normal or preeclamptic PDMSCs.

monoclonal anti-human MIF
  • MIF and sFlt-1 mRNA (left panels) and protein (right panels) expression levels in physiological villous explants treated with unconditioned media [C] or media conditioned by normal [N-cm] and preeclamptic [PE-cm] PDMSCs as assessed by Real Time PCR and Western Blot analysis.