Revision 1

#60125Store at -20C

1 Kit

(6 x 20 microliters)

Cell Signaling Technology

Orders: 877-616-CELL (2355) [email protected]

Support: 877-678-TECH (8324)

Web: [email protected] cellsignal.com

3 Trask LaneDanversMassachusetts01923USA
For Research Use Only. Not for Use in Diagnostic Procedures.
Product Includes Product # Quantity Mol. Wt Isotype/Source
RNase L (D4B4J) Rabbit mAb 27281 20 µl 80 kDa Rabbit IgG
OAS1 (D1W3A) Rabbit mAb 14498 20 µl 40, 44 kDa Rabbit IgG
OAS2 (E2G4K) Rabbit mAb 24344 20 µl 70 kDa Rabbit IgG
OAS3 (E4X7Z) Rabbit mAb 74906 20 µl 120 kDa Rabbit IgG
OASL (E7W1R) Rabbit mAb 36845 20 µl 52 kDa Rabbit IgG
PDE12 (E2V7J) Rabbit mAb 88499 20 µl 75, 70 kDa Rabbit IgG
Anti-rabbit IgG, HRP-linked Antibody 7074 100 µl Goat 

Please visit cellsignal.com for individual component applications, species cross-reactivity, dilutions, protocols, and additional product information.

Description

The OAS/RNase L Pathway Antibody Sampler Kit provides an economical means of detecting proteins involved in the OAS/RNase L pathway. The kit includes enough antibodies to perform two western blot experiments with each primary antibody.

Storage

Supplied in 10 mM sodium HEPES (pH 7.5), 150 mM NaCl, 100 µg/mL BSA, 50% glycerol, and less than 0.02% sodium azide. Store at –20°C. Do not aliquot the antibody.

Background

The 2'-5'-oligoadenylate synthetase (OAS)/RNase L pathway is a component of the antiviral innate immune system. The OAS family of proteins includes OAS1, OAS2, OAS3, and OASL in humans, all of which are induced in response to type 1 interferons (1-3). When activated by cytosolic double-stranded RNA, OAS1-3 generate 2’-5’-linked oligoadenylates that bind to and activate the ribonuclease, RNase L. Activated RNase L is poised to degrade viral and cellular RNA as part of the antiviral innate immune response (4). Unlike OAS1-3, OASL lacks intrinsic enzymatic activity, and research studies have suggested that OASL participates in the antiviral immune response independently of the canonical OAS/RNase L signaling pathway (5,6). Negative regulation of the OAS/RNase L pathway is governed, in part, by the PDE12 phosphodiesterase that degrades 2'-5'-oligoadenylates (7).

  1. Chebath, J. et al. Nature 330, 587-8.
  2. Schoggins, J.W. et al. (2011) Nature 472, 481-5.
  3. (2019) Proc Natl Acad Sci U S A 116, 12573.
  4. Dong, B. and Silverman, R.H. (1997) J Biol Chem 272, 22236-42.
  5. Marques, J. et al. (2008) J Gen Virol 89, 2767-2772.
  6. Ishibashi, M. et al. (2010) Biochem Biophys Res Commun 392, 397-402.
  7. Wood, E.R. et al. (2015) J Biol Chem 290, 19681-96.

Background References

    Trademarks and Patents

    Cell Signaling Technology is a trademark of Cell Signaling Technology, Inc.
    All other trademarks are the property of their respective owners. Visit cellsignal.com/trademarks for more information.

    使用に関する制限

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    Revision 1
    #60125

    OAS/RNase L Pathway Antibody Sampler Kit

    OAS/RNase L Pathway Antibody Sampler Kit: Image 1 Expand Image

    Simple Western™ analysis of HT-29 + hIFN⍺ -1 (10ng/mL, O/N) + poly(I:C )(2.5µg/mL, 7hr) + MG-132 (10µM, 7hr) lysates (0.1 mg/mL) using OAS1 (D1W3A) Rabbit mAb #14498. The virtual lane view (left) shows the target band (as indicated) at 1:10 and 1:50 dilutions of primary antibody. The corresponding electropherogram view (right) plots chemiluminescence by molecular weight along the capillary at 1:10 (blue line) and 1:50 (green line) dilutions of primary antibody. This experiment was performed under reducing conditions on the Jess Simple Western instrument from ProteinSimple, a BioTechne brand, using the 12-230 kDa separation module.

    OAS/RNase L Pathway Antibody Sampler Kit: Image 2 Expand Image
    Western blot analysis of extracts from various cell lines using OAS3 (E4X7Z) Rabbit mAb (upper) or GAPDH (D16H11) XP® Rabbit mAb #5174 (lower). Negative expression of OAS3 protein in 293T and Raji cells is consistent with the predicted expression pattern.
    OAS/RNase L Pathway Antibody Sampler Kit: Image 3 Expand Image
    Western blot analysis of extracts from HeLa, NCI-H460, and HT-29 cells, untreated (-) or treated with Human Interferon-α1 (hIFN-α1) #8927 (10 ng/ml, 16 hr; +), using OAS1 (D1W3A) Rabbit mAb (upper) and β-Actin (D6A8) Rabbit mAb #8457 (lower).
    OAS/RNase L Pathway Antibody Sampler Kit: Image 4 Expand Image
    Western blot analysis of extracts from HT-1080 and A549 cells, untreated (-) or treated with Human Interferon-α1 (hIFN-α1) #8927 (100 ng/ml, 24 hr; +), using OAS2 (E2G4K) Rabbit mAb (upper) and β-Actin (D6A8) Rabbit mAb #8457 (lower).
    OAS/RNase L Pathway Antibody Sampler Kit: Image 5 Expand Image
    Western blot analysis of extracts from various cell lines using RNase L (D4B4J) Rabbit mAb.
    OAS/RNase L Pathway Antibody Sampler Kit: Image 6 Expand Image
    Western blot analysis of extracts from 293T cells, mock transfected (-) or transfected with a construct expressing Myc/DDK-tagged full-length human OASL protein (hOASL-Myc/DDK; +), using OASL (E7W1R) Rabbit mAb (upper), Myc-Tag (71D10) Rabbit mAb #2278 (middle), or GAPDH (D16H11) XP® Rabbit mAb #5174 (lower).
    OAS/RNase L Pathway Antibody Sampler Kit: Image 7 Expand Image
    After the primary antibody is bound to the target protein, a complex with HRP-linked secondary antibody is formed. The LumiGLO® is added and emits light during enzyme catalyzed decomposition.
    OAS/RNase L Pathway Antibody Sampler Kit: Image 8 Expand Image
    Western blot analysis of extracts from 293T cells, mock transfected (-) or transfected with a construct expressing Myc/DDK-tagged full-length human OAS3 protein (hOAS3-Myc/DDK; +), using OAS3 (E4X7Z) Rabbit mAb (upper), Myc-Tag (71D10) Rabbit mAb #2278 (middle), or GAPDH (D16H11) XP® Rabbit mAb #5174 (lower).
    OAS/RNase L Pathway Antibody Sampler Kit: Image 9 Expand Image
    Western blot analysis of extracts from various human cell lines using PDE12 (E2V7J) Rabbit mAb.
    OAS/RNase L Pathway Antibody Sampler Kit: Image 10 Expand Image
    Western blot analysis of extracts from 293T cells, mock transfected (-) or transfected with a construct expressing human OAS1 isoform p41 (hOAS1 p41; +), using OAS1 (D1W3A) Rabbit mAb.
    OAS/RNase L Pathway Antibody Sampler Kit: Image 11 Expand Image
    Western blot analysis of extracts from 293T cells, mock transfected (-) or transfected with a construct expressing Myc/DDK-tagged full-length human OAS2 protein (hOAS2-Myc/DDK; +), using OAS2 (E2G4K) Rabbit mAb (upper), DYKDDDDK Tag Antibody #2368 (middle), and β-Actin (D6A8) Rabbit mAb #8457 (lower).
    OAS/RNase L Pathway Antibody Sampler Kit: Image 12 Expand Image
    Western blot analysis of extracts from 293T cells, untransfected (-) or transfected with a construct expressing Myc/DDK-tagged full-length human RNase L (hRNase L-Myc/DDK; +), using RNase L (D4B4J) Rabbit mAb.
    OAS/RNase L Pathway Antibody Sampler Kit: Image 13 Expand Image
    Western blot analysis of extracts from various cell lines using OASL (E7W1R) Rabbit mAb (upper) or β-Actin (D6A8) Rabbit mAb #8457 (lower).
    OAS/RNase L Pathway Antibody Sampler Kit: Image 14 Expand Image
    Western blot analysis of extracts from various human and mouse cell lines using PDE12 (E2V7J) Rabbit mAb.
    OAS/RNase L Pathway Antibody Sampler Kit: Image 15 Expand Image
    Immunoprecipitation of OAS2 protein from NCI-H1650 cell extracts. Lane 1 is 10% input, lane 2 is Rabbit (DA1E) mAb IgG XP® Isotype Control #3900, and lane 3 is OAS2 (E2G4K) Rabbit mAb. Western blot analysis was performed using OAS2 (E2G4K) Rabbit mAb. Anti-rabbit IgG, HRP-linked Antibody #7074 was used as the secondary antibody.
    OAS/RNase L Pathway Antibody Sampler Kit: Image 16 Expand Image
    Western blot analysis of extracts from THP-1, HeLa, and A549 cells, untreated (-) or treated with Human Interferon-α1 (hIFN-α1) #8927 (100 ng/mL, 24 hr; +), using OASL (E7W1R) Rabbit mAb (upper) or β-Actin (D6A8) Rabbit mAb #8457 (lower).
    OAS/RNase L Pathway Antibody Sampler Kit: Image 17 Expand Image
    Western blot analysis of extracts from 293T cells, transfected with control siRNA (-) or PDE12 siRNA (+), using PDE12 (E2V7J) Rabbit mAb (upper) or β-Actin (D6A8) Rabbit mAb #8457 (lower).
    OAS/RNase L Pathway Antibody Sampler Kit: Image 18 Expand Image
    Western blot analysis of extracts from 293T cells, mock transfected (-) or transfected with a construct expressing Myc/DDK-tagged full-length human PDE12 (hPDE12-Myc/DDK; +), using PDE12 (E2V7J) Rabbit mAb (upper), DYKDDDDK Tag (D6W5B) Rabbit mAb #14793 (middle), or β-Actin (D6A8) Rabbit mAb #8457 (lower).