Revision 1

#8674Store at -20C

1 Kit

(8 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
COX IV (3E11) Rabbit mAb 4850 20 µl 17 kDa Rabbit IgG
Cytochrome c (136F3) Rabbit mAb 4280 20 µl 14 kDa Rabbit IgG
HSP60 (D6F1) XP® Rabbit mAb 12165 20 µl 60 kDa Rabbit IgG
PHB1 Antibody 2426 20 µl 29 kDa Rabbit 
Pyruvate Dehydrogenase (C54G1) Rabbit mAb 3205 20 µl 43 kDa Rabbit IgG
SDHA (D6J9M) XP® Rabbit mAb  11998 20 µl 70 kDa Rabbit IgG
SOD1 (71G8) Mouse mAb 4266 20 µl 18 kDa Mouse IgG1
VDAC (D73D12) Rabbit mAb 4661 20 µl 32 kDa Rabbit IgG
Anti-mouse IgG, HRP-linked Antibody 7076 100 µl Horse 
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 Mitochondrial Marker Antibody Sampler Kit provides an economical means to evaluate relevant mitochondial proteins. This kit contains enough primary antibody to perform two western blots per primary.

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 Mitochondrial Marker Antibody Sampler Kit contains a variety of antibodies directed against established mitochondrial proteins. Cytochrome c oxidase (COX) is a hetero-oligomeric enzyme consisting of 13 subunits localized to the inner mitochondrial membrane (1). Cytochrome c is a well conserved electron-transport protein and is part of the respiratory chain localized to the mitochondrial intermembrane space (2). HSP60 has primarily been known as a mitochondrial protein that is important for folding key proteins after import into the mitochondria (3). In the mitochondria, prohibitins (PHB1) mainly exist as membrane-bound ring complexes and function as chaperones maintaining mitochondrial protein stability during protein synthesis and transportation (4). In mammalian cells, the pyruvate dehydrogenase complex is located in the mitochondrial matrix (5). Succinate dehydrogenase (SDH), also known as Complex II or succinate quinone oxidoreductase, is a key component of the citric acid cycle and the electron transport chain (6). SOD1 is ubiquitously expressed and is localized in the cytosol, nucleus, and mitochondrial intermembrane space (7). Voltage-dependent anion channel (VDAC), ubiquitously expressed and located in the outer mitochondrial membrane, is generally thought to be the primary means by which metabolites diffuse in and out of the mitochondria (8).

  1. Ostermeier, C. et al. (1996) Curr Opin Struct Biol 6, 460-6.
  2. Schägger, H. (2002) Biochim Biophys Acta 1555, 154-9.
  3. Jindal, S. et al. (1989) Mol Cell Biol 9, 2279-83.
  4. Tatsuta, T. et al. (2005) Mol Biol Cell 16, 248-59.
  5. Strumiło, S. (2005) Acta Biochim Pol 52, 759-64.
  6. Oyedotun, K.S. and Lemire, B.D. (2004) J Biol Chem 279, 9424-31.
  7. Sherman, L. et al. (1983) Proc Natl Acad Sci U S A 80, 5465-9.
  8. Craigen, W.J. and Graham, B.H. (2008) J Bioenerg Biomembr 40, 207-12.

Background References

    Trademarks and Patents

    Cell Signaling Technology is a trademark of Cell Signaling Technology, Inc.
    U.S. Patent No. 7,429,487, foreign equivalents, and child patents deriving therefrom.
    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 1
    #8674

    Mitochondrial Marker Antibody Sampler Kit

    Mitochondrial Marker Antibody Sampler Kit: Image 1 Expand Image
    Immunoprecipitation of SOD1 from MCF7 cell extracts. Lane 1 is 10% input, lane 2 is precipitated with Mouse (G3A1) mAb IgG1 Isotype Control #5415, and lane 3 is SOD1 (71G8) Mouse mAb #4266. Western blot was performed using SOD1 (E4G1H) XP® Rabbit mAb #37385, to prevent heavy and light chain IgG masking.
    Mitochondrial Marker Antibody Sampler Kit: Image 2 Expand Image
    Simple Western analysis of lysates (1.0 mg/mL) from COS-7 cells using COX IV (3E11) Rabbit mAb #4850. The virtual lane view (left) shows the target bands (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.
    Mitochondrial Marker Antibody Sampler Kit: Image 3 Expand Image
    Western blot analysis of extracts from various cell lines using SDHA (D6J9M) XP® Rabbit mAb.
    Mitochondrial Marker Antibody Sampler Kit: Image 4 Expand Image
    Western blot analysis of extracts from various cell lines using HSP60 (D6F1) XP® Rabbit mAb.
    Mitochondrial Marker Antibody Sampler Kit: Image 5 Expand Image
    Western blot analysis of extracts from MCF-7, Neuro2A, PC12 and COS cells, using PHB1 Antibody.
    Mitochondrial Marker Antibody Sampler Kit: Image 6 Expand Image
    Western blot analysis of extracts from various cell lines using Pyruvate Dehydrogenase (C54G1) Rabbit mAb.
    Mitochondrial Marker Antibody Sampler Kit: Image 7 Expand Image
    Western blot analysis of extracts from various cell lines using SOD1 (71G8) Mouse mAb.
    Mitochondrial Marker Antibody Sampler Kit: Image 8 Expand Image
    Western blot analysis of cell lysates from various cell lines, using Cytochrome c (136F3) Rabbit mAb.
    Mitochondrial Marker Antibody Sampler Kit: Image 9 Expand Image
    Western blot analysis of extracts from various cell types using VDAC (D73D12) Rabbit mAb.
    Mitochondrial Marker Antibody Sampler Kit: Image 10 Expand Image
    Western blot analysis of extracts from HeLa, Jurkat and COS cell lines, using COX IV (3E11) Rabbit mAb.
    Mitochondrial Marker Antibody Sampler Kit: Image 11 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.
    Mitochondrial Marker Antibody Sampler Kit: Image 12 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.
    Mitochondrial Marker Antibody Sampler Kit: Image 13 Expand Image
    Immunoprecipitation of SDHA from HeLa cell extracts using Rabbit (DA1E) mAb IgG XP® Isotype Control #3900 (lane 2) or SDHA (D6J9M) XP® Rabbit mAb (lane 3). Lane 1 is 10% input. Western blot analysis was performed using SDHA (D6J9M) XP® Rabbit mAb.
    Mitochondrial Marker Antibody Sampler Kit: Image 14 Expand Image
    Immunohistochemical analysis of paraffin-embedded mouse lung using HSP60 (D6F1) XP® Rabbit mAb.
    Mitochondrial Marker Antibody Sampler Kit: Image 15 Expand Image
    Western blot analysis of various cell lines using Pyruvate Dehydrogenase (C54G1) Rabbit mAb.
    Mitochondrial Marker Antibody Sampler Kit: Image 16 Expand Image
    Immunohistochemical analysis of paraffin-embedded human colon carcinoma, using Cytochrome c (136F3) Rabbit mAb.
    Mitochondrial Marker Antibody Sampler Kit: Image 17 Expand Image
    Immunohistochemical analysis of paraffin-embedded human colon carcinoma using VDAC (D73D12) Rabbit mAb.
    Mitochondrial Marker Antibody Sampler Kit: Image 18 Expand Image
    Immunohistochemical analysis of paraffin-embedded human colon carcinoma, showing staining of the mitochondria, using COX IV (3E11) Rabbit mAb.
    Mitochondrial Marker Antibody Sampler Kit: Image 19 Expand Image
    Immunohistochemical analysis of paraffin-embedded human colon carcinoma using SDHA (D6J9M) XP® Rabbit mAb in the presence of control peptide (left) or antigen-specific peptide (right).
    Mitochondrial Marker Antibody Sampler Kit: Image 20 Expand Image
    Immunohistochemical analysis of paraffin-embedded human breast carcinoma using HSP60 (D6F1) XP® Rabbit mAb.
    Mitochondrial Marker Antibody Sampler Kit: Image 21 Expand Image
    Immunohistochemical analysis of paraffin-embedded human lung carcinoma using Pyruvate Dehydrogenase (C54G1) Rabbit mAb.
    Mitochondrial Marker Antibody Sampler Kit: Image 22 Expand Image
    Immunohistochemical analysis of paraffin-embedded 4T1 syngeneic mouse tumor, using Cytochrome c (136F3) Rabbit mAb #4280.
    Mitochondrial Marker Antibody Sampler Kit: Image 23 Expand Image
    Immunohistochemical analysis of paraffin-embedded human esophageal carcinoma using VDAC (D73D12) Rabbit mAb.
    Mitochondrial Marker Antibody Sampler Kit: Image 24 Expand Image
    Immunohistochemical analysis of paraffin-embedded human breast carcinoma, using COX IV (3E11) Rabbit mAb in the presence of control peptide (left) or Cox IV Blocking Peptide #1034 (right).
    Mitochondrial Marker Antibody Sampler Kit: Image 25 Expand Image
    Confocal immunofluorescent analysis of HeLa cells using SDHA (D6J9M) XP® Rabbit mAb (green) and β-Actin (8H10D10) Mouse mAb #3700 (red). Blue pseudocolor = DRAQ5® #4084 (fluorescent DNA dye).
    Mitochondrial Marker Antibody Sampler Kit: Image 26 Expand Image
    Immunohistochemical analysis of paraffin-embedded human ovarian carcinoma using HSP60 (D6F1) XP® Rabbit mAb in the presence of control peptide (left) or antigen-specific peptide (right).
    Mitochondrial Marker Antibody Sampler Kit: Image 27 Expand Image
    Immunohistochemical analysis of paraffin-embedded human breast carcinoma using Pyruvate Dehydrogenase (C54G1) Rabbit mAb in the presence of control peptide (left) or antigen-specific peptide (right).
    Mitochondrial Marker Antibody Sampler Kit: Image 28 Expand Image
    Immunohistochemical analysis of paraffin-embedded human lung carcinoma, using Cytochrome c (136F3) Rabbit mAb in the presence of control peptide (left) or Cytochrome c Blocking Peptide #1033 (right).
    Mitochondrial Marker Antibody Sampler Kit: Image 29 Expand Image
    Immunohistochemical analysis of paraffin-embedded human squamous cell lung carcinoma using VDAC (D73D12) Rabbit mAb.
    Mitochondrial Marker Antibody Sampler Kit: Image 30 Expand Image
    Immunohistochemical analysis of paraffin-embedded H1650 xenograft, using COX IV Rabbit mAb. Note specific staining of human cancer cells.
    Mitochondrial Marker Antibody Sampler Kit: Image 31 Expand Image
    Confocal immunofluorescent analysis of A-204 cells using HSP60 (D6F1) XP® Rabbit mAb (green). Actin filaments were labeled with DY-554 phalloidin (red). Blue pseudocolor = DRAQ5® #4084 (fluorescent DNA dye).
    Mitochondrial Marker Antibody Sampler Kit: Image 32 Expand Image
    Immunohistochemical analysis of paraffin-embedded human prostate carcinoma, using Cytochrome c (136F3) Rabbit mAb .
    Mitochondrial Marker Antibody Sampler Kit: Image 33 Expand Image
    Immunohistochemical analysis of paraffin-embedded human prostate carcinoma using VDAC (D73D12) Rabbit mAb.
    Mitochondrial Marker Antibody Sampler Kit: Image 34 Expand Image
    Flow cytometric analysis of HeLa cells using HSP60 (D6F1) XP® Rabbit mAb (blue) compared to concentration-matched Rabbit (DA1E) mAb IgG XP® Isotype Control #3900 (red). Anti-rabbit IgG (H+L), F(ab')2 Fragment (Alexa Fluor® 488 Conjugate) #4412 was used as a secondary antibody.
    Mitochondrial Marker Antibody Sampler Kit: Image 35 Expand Image
    Immunohistochemical analysis of paraffin-embedded mouse skeletal muscle using VDAC (D73D12) Rabbit mAb.
    Mitochondrial Marker Antibody Sampler Kit: Image 36 Expand Image
    Confocal immunofluorescent analysis of HeLa cells labeled with COX IV (3E11) Rabbit mAb (green) and β-Actin (8H10D10) Mouse mAb #3700 (red). Samples were mounted in ProLong® Gold Antifade Reagent with DAPI #8961 (blue).
    Mitochondrial Marker Antibody Sampler Kit: Image 37 Expand Image
    Immunohistochemical analysis of paraffin-embedded mouse small intestine using VDAC (D73D12) Rabbit mAb.
    Mitochondrial Marker Antibody Sampler Kit: Image 38 Expand Image
    Flow cytometric analysis of HeLa cells using COX IV (3E11) Rabbit mAb (solid line) compared to concentration-matched Rabbit (DA1E) mAb IgG XP® Isotype Control #3900 (dashed line). Anti-rabbit IgG (H+L), F(ab')₂ Fragment (Alexa Fluor® 488 Conjugate) #4412 was used as a secondary antibody.