Revision 5

#73959Store at -20C

1 Kit

(8 x 20 microliters)

Cell Signaling Technology

Orders: 877-616-CELL (2355) orders@cellsignal.com

Support: 877-678-TECH (8324)

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3 Trask LaneDanversMassachusetts01923USA
For Research Use Only. Not for Use in Diagnostic Procedures.
Product Includes Product # Quantity Mol. Wt Isotype/Source
MMP-9 (D6O3H) XP® Rabbit mAb 13667 20 µl 84, 92 kDa Rabbit IgG
MMP-2 (D2O4T) Rabbit mAb 87809 20 µl 64,72 kDa Rabbit IgG
MMP-3 (D7F5B) Rabbit mAb 14351 20 µl 60 kDa Rabbit IgG
TIMP1 (D10E6) Rabbit mAb 8946 20 µl 26 kDa Rabbit IgG
TIMP2 (D18B7) Rabbit mAb 5738 20 µl 22 kDa Rabbit IgG
TIMP3 (D74B10) Rabbit mAb 5673 20 µl 20, 25 kDa Rabbit IgG
MMP-7 Antibody 71031 20 µl 28 kDa Rabbit 
MT1-MMP (E3S5S) Rabbit mAb 26424 20 µl 62 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 Matrix Remodeling Antibody Sampler Kit provides an economical means of detecting different MMPs and TIMPs using the specific corresponding antibodies. The kit contains enough antibody to perform at least 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

Matrix remodeling is mainly controlled by MMPs and TIMPs. The matrix metalloproteinase (MMP) family of proteases are a group of zinc-dependent enzymes that target extracellular proteins, including growth factors, cell surface receptors, adhesion molecules, matrix structural proteins, and other proteases (1, 2). Among the family members, MMP-2, MMP-3, MMP-7, MMP-9, and MMP14 (MT1-MMP) have been characterized as important factors for normal tissue remodeling during embryonic development, wound healing, tumor invasion, angiogenesis, carcinogenesis, and apoptosis (3). MMP activity is regulated by mechanisms of both transcriptional control and post translational protein processing. Once synthesized, MMPs exist as latent proenzymes. Maximum MMP activity requires proteolytic cleavage to generate active MMPs by releasing the inhibitory propeptide domain from the full-length protein (4). MMP activity can be inhibited through its binding to endogenously expressed TIMPs. TIMPs are members of the family of tissue inhibitors of matrix metalloproteinases that include TIMP1, TIMP2, TIMP3, and TIMP4. The main function of TIMPs is their inhibitory effect on MMPs. TIMPs irreversibly inactivate MMPs by direct binding MMPs and chelating their zinc cofactor at the catalytic site to inhibit the proteinase function (5,6).

  1. Kessenbrock, K. et al. (2010) Cell 141, 52-67.
  2. McCawley, L.J. and Matrisian, L.M. (2001) Curr Opin Cell Biol 13, 534-40.
  3. Page-McCaw, A. et al. (2007) Nat Rev Mol Cell Biol 8, 221-33.
  4. Hadler-Olsen, E. et al. (2011) FEBS J 278, 28-45.
  5. Nagase, H. et al. (2006) Cardiovasc Res 69, 562-73.
  6. Visse, R. and Nagase, H. (2003) Circ Res 92, 827-39.

Background References

    Trademarks and Patents

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

    Limited Uses

    Except as otherwise expressly agreed in a writing signed by a legally authorized representative of CST, the following terms apply to Products provided by CST, its affiliates or its distributors. Any Customer's terms and conditions that are in addition to, or different from, those contained herein, unless separately accepted in writing by a legally authorized representative of CST, are rejected and are of no force or effect.

    Products are labeled with For Research Use Only or a similar labeling statement and have not been approved, cleared, or licensed by the FDA or other regulatory foreign or domestic entity, for any purpose. Customer shall not use any Product for any diagnostic or therapeutic purpose, or otherwise in any manner that conflicts with its labeling statement. Products sold or licensed by CST are provided for Customer as the end-user and solely for research and development uses. Any use of Product for diagnostic, prophylactic or therapeutic purposes, or any purchase of Product for resale (alone or as a component) or other commercial purpose, requires a separate license from CST. Customer shall (a) not sell, license, loan, donate or otherwise transfer or make available any Product to any third party, whether alone or in combination with other materials, or use the Products to manufacture any commercial products, (b) not copy, modify, reverse engineer, decompile, disassemble or otherwise attempt to discover the underlying structure or technology of the Products, or use the Products for the purpose of developing any products or services that would compete with CST products or services, (c) not alter or remove from the Products any trademarks, trade names, logos, patent or copyright notices or markings, (d) use the Products solely in accordance with CST Product Terms of Sale and any applicable documentation, and (e) comply with any license, terms of service or similar agreement with respect to any third party products or services used by Customer in connection with the Products.

    Revision 5
    #73959

    Matrix Remodeling Antibody Sampler Kit

    Matrix Remodeling Antibody Sampler Kit: Image 1 Expand Image

    Simple Western™ analysis of lysates (1.0 mg/mL) from 3T3-L1 cells using MMP-2 (D2O4T) Rabbit mAb #87809. 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.

    Matrix Remodeling Antibody Sampler Kit: Image 2 Expand Image
    Western blot analysis of concentrated, serum-free cultured medium from U-2 OS cells, untreated (-) or treated with TPA #4174 (200 nM, 48 hr; +), using MMP-9 (D6O3H) XP® Rabbit mAb.
    Matrix Remodeling Antibody Sampler Kit: Image 3 Expand Image
    Western blot analysis of extracts from various cell lines using MMP-3 (D7F5B) Rabbit mAb.
    Matrix Remodeling Antibody Sampler Kit: Image 4 Expand Image
    Western blot analysis of extracts from various cell lines using MT1-MMP (E3S5S) Rabbit mAb (upper) or GAPDH (D16H11) XP® Rabbit mAb #5174 (lower). Negative expression of MT1-MMP protein in HCC1419 cells is consistent with the predicted expression pattern.
    Matrix Remodeling Antibody Sampler Kit: Image 5 Expand Image
    Western blot analysis of extracts from A-431, and NIH/3T3 cells, using TIMP3 (D74B10) Rabbit mAb.
    Matrix Remodeling Antibody Sampler Kit: Image 6 Expand Image
    Western blot analysis of extracts from HT-1080, A172, and COS-7 cells using TIMP2 (D18B7) Rabbit mAb.
    Matrix Remodeling 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.
    Matrix Remodeling Antibody Sampler Kit: Image 8 Expand Image
    Western blot analysis of extracts from concentrated culture medium of IGROV-1 and HT-29 cell lines using MMP-7 Antibody.
    Matrix Remodeling Antibody Sampler Kit: Image 9 Expand Image
    Western blot analysis of extracts from U87, 3T3-L1 and C2C12 cells using MMP-2 (D4O4T) Rabbit mAb.
    Matrix Remodeling Antibody Sampler Kit: Image 10 Expand Image
    Western blot analysis of extracts from various cell lines using TIMP1 (D10E6) Rabbit mAb.
    Matrix Remodeling Antibody Sampler Kit: Image 11 Expand Image
    Immunohistochemical analysis of paraffin-embedded human colon carcinoma using TIMP1 (D10E6) Rabbit mAb.
    Matrix Remodeling Antibody Sampler Kit: Image 12 Expand Image
    Western blot analysis of extracts and concentrated culture medium from U-2 OS cells, untreated (-) or treated with TPA #4174 (200 nM, 48 hr; +) using MMP-9 (D6O3H) XP® Rabbit mAb. MMP-9 is induced by TPA treatment as expected.
    Matrix Remodeling Antibody Sampler Kit: Image 13 Expand Image
    Immunoprecipitation of MMP-2 protein from U87 cell extracts. Lane 1 is 10% input, lane 2 is Rabbit (DA1E) mAb IgG XP® Isotype Control #3900, and lane 3 is MMP-2 (D2O4T) Rabbit mAb. Western blot analysis was performed using MMP-2 (D2O4T) Rabbit mAb.
    Matrix Remodeling Antibody Sampler Kit: Image 14 Expand Image
    Immunohistochemical analysis of paraffin-embedded human ductal breast carcinoma using TIMP1 (D10E6) Rabbit mAb.
    Matrix Remodeling Antibody Sampler Kit: Image 15 Expand Image
    Immunohistochemical analysis of paraffin-embedded human breast carcinoma using MMP-9 (D6O3H) XP® Rabbit mAb.
    Matrix Remodeling Antibody Sampler Kit: Image 16 Expand Image
    Immunohistochemical analysis of paraffin-embedded human esophageal caricinoma using TIMP1 (D10E6) Rabbit mAb.

    Matrix Remodeling Antibody Sampler Kit: Image 17 Expand Image
    Immunohistochemical analysis of paraffin-embedded human lung carcinoma using MMP-9 (D6O3H) XP® Rabbit mAb.
    Matrix Remodeling Antibody Sampler Kit: Image 18 Expand Image
    Immunohistochemical analysis of paraffin-embedded human squamous cell lung carcinoma using TIMP1 (D10E6) Rabbit mAb.
    Matrix Remodeling Antibody Sampler Kit: Image 19 Expand Image
    Immunohistochemical analysis of paraffin-embedded U-2 OS cell pellets, untreated (left) or treated with TPA #4174 (right), using MMP-9 (D6O3H) XP® Rabbit mAb.
    Matrix Remodeling Antibody Sampler Kit: Image 20 Expand Image
    Immunohistochemical analysis of paraffin-embedded normal human pancreas using TIMP1 (D10E6) Rabbit mAb.
    Matrix Remodeling Antibody Sampler Kit: Image 21 Expand Image
    Immunohistochemical analysis of paraffin-embedded human colon carcinoma using MMP-9 (D6O3H) XP® Rabbit mAb in the presence of control peptide (left) or antigen-specific peptide (right).
    Matrix Remodeling Antibody Sampler Kit: Image 22 Expand Image
    Immunohistochemical analysis of paraffin-embedded normal human small intestine using TIMP1 (D10E6) Rabbit mAb.
    Matrix Remodeling Antibody Sampler Kit: Image 23 Expand Image
    Immunohistochemical analysis of paraffin-embedded human prostate carcinoma using TIMP1 (D10E6) Rabbit mAb (left) compared to concentration-matched Rabbit (DA1E) mAb IgG XP® Isotype Control #3900 (right).
    Matrix Remodeling Antibody Sampler Kit: Image 24 Expand Image
    Flow cytometric analysis of U-2 OS cells, untreated (blue) or treated with TPA (12-O-Tetradecanoylphorbol-13-Acetate) #4174 (200 nM, 24 hr; green) using MMP-9 (D6O3H) XP® Rabbit mAb (solid lines) or concentration-matched Rabbit (DA1E) mAb IgG XP® Isotype Control #3900 (dashed lines). Anti-rabbit IgG (H+L), F(ab')2 Fragment (Alexa Fluor® 488 Conjugate) #4412 was used as a secondary antibody.
    Matrix Remodeling Antibody Sampler Kit: Image 25 Expand Image
    Immunohistochemical analysis of paraffin-embedded U-87 MG cell pellet (left, positive) or Ramos cell pellet (right, negative) using TIMP1 (D10E6) Rabbit mAb.