dsRNA Antibody Pack

• Reactivity: All-Multi
• Applications: ELISA, Flow, ICC/IF, IHC, WB, AP
• Clonality: Monoclonal

dsRNA Antibody Pack Summary

• Description: This pack contains 1 vial each of: NBP3-11395 (100 ug); NBP3-11396 (100 ug); NBP3-11397 (100 ug). This ds-RNA antibody was manufactured by Scicon.
• Immunogen: Female DBA/2 mice were injected intraperitonially with a mixture of 50 ug L-dsRNA and 75 ug methylated bovine serum albumin, emulsified in complete Freund's adjuvant. After several boosts spleen cells were fused with Sp2/0-Agl4 myeloma cells to generate the hybridoma clones.
• Specificity: Mouse monoclonal antibodies NBP3-11395, NBP3-11396, and NBP3-11397 recognise double-stranded RNA (dsRNA) provided that the length of the helix is greater than or equal to 40 bp. dsRNA-recognition is independent of the sequence and nucleotide composition of the antigen. All naturally occurring dsRNAs investigated up to now (40-50 species) as well as poly(I).poly(C) and poly(A).poly(U) have been recognised by all three antibodies, although in some assays the affinity of NBP3-11395 and NBP3-11396 to poly(I).poly(C) is about 10 times lower than that to other dsRNA antigens. NBP3-11397 shows higher affinity to poly(I).poly(C) than to the other dsRNA antigens, although the difference of apparent binding constants may vary under different experimental conditions.
• Clonality: Monoclonal
• Purity: Protein A purified

Applications/Dilutions

• Dilutions:
  • ELISA
  • Flow Cytometry
  • Immunoaffinity Purification
  • Immunoblotting
  • Immunocytochemistry/ Immunofluorescence
  • Immunohistochemistry
• Application Notes: Antibodies in this pack are validated for the following applications:
  
  NBP3-11397: ELISA, Flow Cytometry, Immunoaffinity Purification, Immunoblotting, Immunocytochemistry/Immunofluorescence, Immunohistochemistry
  NBP3-11395: ELISA, Immunoaffinity Purification, Immunocytochemistry/Immunofluorescence, Immunohistochemistry, Western Blot
  NBP3-11396: ELISA, Immunoblotting, Immunocytochemistry/Immunofluorescence, Immunohistochemistry
  NL007: Flow Cytometry, Immunocytochemistry, Immunohistochemistry
  
  Please note that nucleic acid separation prior to dsRNA-immunoblotting must be carried out by polyacrylamide gel electrophoresis, because the sensitivity of detection is considerably lower after blotting from agarose gels.
  
  As a result of the lyophilisation procedure, the reconstituted antibody may contain small amounts of denatured protein in the form of aggregates that may interfere with some applications such as immunohistochemistry (e.g. by giving high backgrounds). We therefore highly recommend centrifuging (microcentrifuge) the reconstituted antibody before use and using the supernatant..

Reactivity Notes

Antibodies in this pack are validated for use in the following species:

NBP3-11397: Multi-Species
NBP3-11395: Multi-Species
NBP3-11396: Multi-Species
NL007: Mouse

Packaging, Storage & Formulations

• Storage: Store lyophilized antibody at 4C. Aliquot reconstituted liquid and store at -20C. Avoid freeze-thaw cycles.
• Buffer: After reconstitution the mAbs will be in PBS without any stabilisers or preservatives at a concentration of 1 mg/ml
• Preservative: No Preservative
• Purity: Protein A purified
• Reconstitution Instructions: The lyophilised samples should each be reconstituted with 100 ul sterile distilled water.

Alternate Names for dsRNA Antibody Pack

• Double-stranded RNA

Background

Over the past decade our double-stranded RNA (dsRNA)antibodies have been used extensively to detect and characterise plant and animal viruses with dsRNA genomes or intermediates.

The NBP3-11397 monoclonal antibody recognises dsRNA with similar affinity to our widely used NBP3-11395 antibody. It has proven especially useful as an alternative to NBP3-11395 to resolve cross-reactions and/or remove unwanted background, in those rare experimental setups where NBP3-11395 did not provide satisfactory results. NBP3-11397 can be used to detect dsRNA intermediates of viruses as diverse as Hepatitis virus, Theiler's murine encephalomyelitis virus or Japanese encephalitis virus. If Poly I:C needs to be detected we highly recommend using NBP3-11397 rather than NBP3-11395 because NBP3-11397 has a much higher affinity for this synthetic polyribonucleotide.

The NBP3-11396 IgG2b antibody recognizes dsRNA with very similar affinity and specificity to our NBP3-11395 antibody, but has a different isotype - thus allowing more flexibility for the simultaneous detection of dsRNA with other markers, particularly in immunofluorescence microscopy. The NBP3-11396 antibody can detect all tested forms of dsRNA, including poly(A):poly(U), poly(I):poly(C) and dsRNA from viruses such as Dengue Virus, Encephalomyocarditis Virus, Vaccinia Virus, Reovirus or Cucumber Mosaic Virus. Similarly to our other antibodies dsRNA-binding of NBP3-11396 is sequence-independent, as long as the length of the dsRNA exceeds 40nt. The antibody does not react with ssRNA, ssDNA or dsDNA.

Limitations

This product is for research use only and is not approved for use in humans or in clinical diagnosis. Antibody Packs are guaranteed for 1 year from date of receipt.