Cell Signaling Technology

Product Pathways - Lymphocyte Signaling

SignalSilence® β2-microglobulin siRNA II #7774

Applications Reactivity
Transfection H

Reactivity Key:  H=Human
Species cross-reactivity is determined by western blot. Species enclosed in parentheses are predicted to react based on 100% sequence homology.

Western Blotting

Western Blotting

Western blot analysis of extracts from HeLa cells, transfected with 100 nM SignalSilence® Control siRNA (Unconjugated) #6568 (-), SignalSilence® β2-microglobulin siRNA I #7759 (+), or SignalSilence® β2-microglobulin siRNA II (+), using β2-microglobulin Antibody #9899 (upper) or β-Tubulin (9F3) Rabbit mAb #2128 (lower). The β2-microglobulin Antibody confirms silencing of β2-microglobulin expression, while the β-Tubulin (9F3) Rabbit mAb is used as a loading control.

Description

SignalSilence® β2-microglobulin siRNA II from Cell Signaling Technology (CST) allows the researcher to specifically inhibit β2-microglobulin expression using RNA interference, a method whereby gene expression can be selectively silenced through the delivery of double stranded RNA molecules into the cell. All SignalSilence® siRNA products from CST are rigorously tested in-house and have been shown to reduce target protein expression by western analysis.

Quality Control

Oligonucleotide synthesis is monitored base by base through trityl analysis to ensure appropriate coupling efficiency. The oligo is subsequently purified by affinity-solid phase extraction. The annealed RNA duplex is further analyzed by mass spectrometry to verify the exact composition of the duplex. Each lot is compared to the previous lot by mass spectrometry to ensure maximum lot-to-lot consistency.

Directions for Use

CST recommends transfection with 100 nM SignalSilence® β2-microglobulin siRNA II 48 to 72 hours prior to cell lysis. For transfection procedure, follow protocol provided by the transfection reagent manufacturer. Please feel free to contact CST with any questions on use.

Each vial contains the equivalent of 100 transfections, which corresponds to a final siRNA concentration of 100 nM per transfection in a 24-well plate with a total volume of 300 μl per well.

Background

β2-microglobulin (B2M) is a principal component of the Major Histocompatibility Complex (MHC) class I molecule, a ternary membrane protein complex that displays fragments derived from proteolyzed cytosolic proteins on the surface of cells for recognition by the surveillance immune system (1,2). As an integral component of the MHC class I complex, β2-microglobulin plays a critically important role in immune system function (3). It has important relevance to cancer biology research; for example, research studies have shown that nearly one-third of diffuse large B cell lymphomas contain mutations that inactivate β2-microglobulin gene function, thereby allowing tumor cells to escape immune detection (4). In additon, β2-microglobulin has been identified as an amyloid preprotein with collagen-binding affinity (5); its accumulation in osteoarthritic lesions of long-term dialysis patients is reportedly a contributing factor to the condition known as amyloid osteoarthropathy (6).

  1. Krangel, M.S. et al. (1979) Cell 18, 979-91.
  2. Collins, E.J. et al. (1995) Proc Natl Acad Sci U S A 92, 1218-21.
  3. Marx, J.I. (1974) Science 185, 428-9.
  4. Challa-Malladi, M. et al. (2011) Cancer Cell 20, 728-40.
  5. Gorevic, P.D. et al. (1985) J Clin Invest 76, 2425-9.
  6. Ohashi, K. (2001) Pathol Int 51, 1-10.

Application References

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Companion Products

Limited Use Label License, RNA interference: This product is licensed under European Patent 1144623 and foreign equivalents from Ribopharma AG, Kulmbach, Germany and is provided only for use in non-commercial research specifically excluding use (a) in drug discovery or drug development, including target identification or target validation, by or on behalf of a commercial entity, (b) for contract research or commercial screening services, (c) for the production or manufacture of siRNA-related products for sale, or (d) for the generation of commercial databases for sale to Third Parties. Information about licenses for these and other commercial uses is available from Ribopharma AG, Fritz-Hornschuch-Str. 9, D-95326 Kulmbach, Germany.

For Research Use Only. Not For Use In Diagnostic Procedures.

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