Introducing Scicons

Exalpha is excited to announce the addition of SCICONS innovative anti-dsRNA antibodies to its portfolio. SCICONS products are powerful new tools in the field of infectious disease research and exemplify Exalpha’s dedication to accelerating the pace of discovery.

SCICONS Products

SCICONS is a supplier of J2, K1 and K2, three mouse monoclonal antibodies that bind to double-stranded RNA molecules.

Application

The anti-dsRNA antibodies can be used as a diagnostic tool to detect whether an unknown pathogen is bacterial or viral in nature, including detection in paraffin-embedded fixed tissue samples. In addition, the J2 anti-dsRNA monoclonal antibody has become the gold standard for the  immunohistological detection of dsRNA.

 

Mouse anti double-stranded RNA (J2)

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. In addition, the anti-dsRNA antibodies can be used as a diagnostic tool to detect pathogens, including detection in paraffin-embedded fixed tissue samples (Richardson et al. 2010). The J2 anti-dsRNA IgG2a monoclonal antibody has become the gold standard in dsRNA detection. It was used initially for the study of plant viruses, but since the seminal paper of Weber et al. in 2006, where J2 was used to show that all the positive strand RNA viruses tested produced copious amounts of dsRNA in infected cells, this antibody has been used extensively in a wide range of systems, as documented in over 200 scientific publications.

J2 can be used to detect dsRNA intermediates of viruses as diverse as Hepatitis C virus, Dengue virus, rhinovirus, Chikungunya virus, Rabies virus, Polio virus, Classic swine fever virus, Brome mosaic virus and many more in cultured cells and also in fixed paraffin-embedded histological samples. J2 has been used to elucidate how anti-viral responses are initiated, what counter-strategies viruses have adopted to avoid them, and to explore the viral life cylce by enabling ultrastructiural localisation studies of viral nucleic acid replication sites (Welsch et al., 2009 & Knoops et al., 2011). J2 has been used successfully in electron microscopy, in immunofluorescence microscopy, in immunohistochemistry, and various immunocapture methods, such as dot blots and ELISA. J2 has also been recommended as a diagnostic tool to detect whether an unkown pathogen is bacterial or viral in nature (Richardson et al., 2010). Recently J2 has also been used to monitor the removal of dsRNA from in vitro synthethised mRNA preparations that may have potential use in gene therapy (Kariko et al., 2011).

Product

Mouse monoclonal antibody J2 recognises 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 J2, although in some assays its affinity to poly(I).poly(C) is about 10 times lower than that to other dsRNA antigens.

Formulation: The lyophilised sample should be reconstituted with 200 µl sterile distilled water. The mAb will then be in PBS without any stabilisers at a concentration of 1 mgr/ml. 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.

Purification Method: Affinity chromatography on Protein A-agarose.

Purity: Gel electrophoretically pure IgG antibody.

Concentration: Concentration after reconstitution: 1.00 mg/ml as determined by A280 nm (A280 nm = 1.47 corresponds to 1 mg/ml antibody).

Application

Mouse monoclonal antibody J2 can be used for ELISA, dsRNA-immunoblotting, immunoaffinity chromatography and in certain systems also for immunohistochemistry (see references). The optimum working dilution of the antibody for any specific application should be established by titration.

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.

Not for use for clinical purposes. For in vitro use only.

View product here.

Mouse anti double-stranded RNA (K1)

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. In addition, the anti-dsRNA antibodies can be used as a diagnostic tool to detect pathogens, including detection in paraffin-embedded fixed tissue samples (Richardson et al. 2010). The K1 monoclonal antibody recognises dsRNA with similar affinity to our widely used J2 antibody. It can be used for the histological and cytological detection of dsRNA in cells and tissues. It has proven especially useful as an alternative to J2 to resolve cross-reactions and/or remove unwanted background, in those rare experimental setups where J2 did not provide satisfactory results. K1 can be used to detect dsRNA intermediates of viruses as diverse as Hepatitis virus, Theiler’s murine encephalomyelitis virus or Japanese encephalitis virus. It has been for the detection of dsRNA in cultured cells and in fixed paraffin-embedded histological samples (see publications). If Poly I:C needs to be detected we highly using K1 rather than J2 because K1 has a much higher affinity for this synthetic polyribonucleotide (see Schönborn et al. 1991, Fig. 2). K1 has been used successfully in immunofluorescence microscopy, in flow cytometry (FACS) and in immunocapture methods (such as dot-blot and ELISA).

Product

The mAb K1 recognises double-stranded RNA (dsRNA) provided that the length of the helix is greater than or equal to 40 bp. dsRNArecognition 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 K1. As described by Schönborn et al. K1 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.

Formulation: The lyophilised sample should be reconstituted with 200 µl sterile distilled water. The mAb will then be in PBS without any stabilisers or preservatives at a concentration of 1 mgr/ml. 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.

Purification Method: Affinity chromatography on Protein A-agarose.

Purity: Gel electrophoretically pure IgG antibody.

Concentration: Concentration after reconstitution: 1.00 mg/ml as determined by A280 nm (A280 nm = 1.47 corresponds to 1 mg/ml antibody).

 

Application

MAb K1 can be used for ELISA, dsRNA-immunoblotting, immuno-affinity-chromatography and in certain systems also for immunohistochemistry (see references).
The optimum working dilution of the antibody for any specific application should be established by titration.

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.

Not for use for clinical purposes. For in vitro use only.

View product here.

Mouse anti double-stranded RNA (K2)

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. In addition, the anti-dsRNA antibodies can be used as a diagnostic tool to detect pathogens, including detection in paraffin-embedded fixed tissue samples (Richardson et al. 2010). K2 anti-dsRNA monoclonal antibody has an IgM isotype. K2 is mainly used for specialised applications, where J2 and K1 – antibodies that can be shipped at ambient temperature – do not suffice. Due to its IgM isotype K2 is more prone to aggregation at high concentration, and therefore cannot be lyophilised and needs to be shipped on ice. K2 has primarily been used in ELISA and sandwich ELISA. Generally, K2 can be used in applications where an anti-dsRNA antibody with an isotype other than IgG (IgG2a) is required.

 

Product

The mAb K2 recognises 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 (4050 species) as well as poly(I).poly(C) and poly(A).poly(U) have been recognised by K2. As described by Schönborn et al. K2 binds with high avidity to all dsRNAs investigated.

Formulation: Culture supernatant (RPMI, 5% fetal calf serum).

Concentration: Undiluted hybridoma supernatant

Application

MAb K2 is primarily used for a sandwich ELISA to detect and quantitate (after calibration) dsRNA (see Schönborn et al.). For this application it should be diluted 1:2 with PBS. It may also be advantageous to use K2 for immunofluorescence studies. The optimum working dilution of the antibody for any specific application should be established by titration.

Not for use for clinical purposes. For in vitro use only.

View product here.

Caution

This products is intended FOR RESEARCH USE ONLY, and FOR TESTS IN VITRO, not for use in diagnostic or therapeutic procedures involving humans or animals. It may contain hazardous ingredients. Please refer to the Safety Data Sheets (SDS) for additional information and proper handling procedures. Dispose product remainders according to local regulations.This datasheet is as accurate as reasonably achievable, but Exalpha Biologicals accepts no liability for any inaccuracies or omissions in this information.

 

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