Revision 3

#96628Store 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
Tau (D1M9X) XP® Rabbit mAb 46687 20 µl 50-80 kDa Rabbit IgG
Phospho-Tau (Ser202) (D4H7E) Rabbit mAb 39357 20 µl 50-80 kDa Rabbit IgG
Phospho-Tau (Ser396) (PHF13) Mouse mAb 9632 20 µl 50-80 kDa Mouse IgG2b
Phospho-Tau (Ser404) (D2Z4G) Rabbit mAb 20194 20 µl 50-80 kDa Rabbit IgG
Phospho-Tau (Ser404) (D2Z4G) Rabbit mAb 35834 20 µl 50-80 kDa Rabbit IgG
Phospho-Tau (Ser416) (D7U2P) Rabbit mAb 15013 20 µl 50-80 kDa Rabbit IgG
Anti-rabbit IgG, HRP-linked Antibody 7074 100 µl Goat 
Anti-mouse IgG, HRP-linked Antibody 7076 100 µl Horse 

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

Description

The Phospho-Tau Family Antibody Sampler Kit provides an economical means of detecting the activation of Tau family members using phospho-specific and control antibodies. The kit includes enough antibody 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

Tau is a heterogeneous microtubule-associated protein that promotes and stabilizes microtubule assembly, especially in axons. Six isoforms with different amino-terminal inserts and different numbers of tandem repeats near the carboxy terminus have been identified, and tau is hyperphosphorylated at approximately 25 sites by Erk, glycogen synthase kinase-3 (GSK-3), and CDK5 (1,2). Phosphorylation decreases the ability of tau to bind to microtubules. Neurofibrillary tangles are a major hallmark of Alzheimer's disease (AD); these tangles are bundles of paired helical filaments (PHFs) composed of hyperphosphorylated tau. In particular, phosphorylation at Ser396 by GSK-3 or CDK5 destabilizes microtubules. Furthermore, research studies have shown that inclusions of tau are found in a number of other neurodegenerative diseases, collectively known as tauopathies (1,3).

Investigators have shown that Tau phosphorylation at Ser404 destabilizes microtubules and that Tau is hyperphosphorylated at Ser404 in Alzheimer's disease (2,4-6). Research studies indicate that calcium-/calmodulin-dependent protease kinase II (CAM-kkinase II) is responsible for the phosphorylation of Tau at Ser416. Phosphorylated Tau protein is localized with neuronal soma or hippocampal neurons and immortalized GnRH neurons (7). Investigators have shown that Tau is phosphorylated during development and hyperphosphorylated at Ser202 in Alzheimer's disease (8).

  1. Johnson, G.V. and Stoothoff, W.H. (2004) J Cell Sci 117, 5721-9.
  2. Hanger, D.P. et al. (1998) J Neurochem 71, 2465-76.
  3. Bramblett, G.T. et al. (1993) Neuron 10, 1089-99.
  4. Shiurba, R.A. et al. (1996) Brain Res 737, 119-32.
  5. Evans, D.B. et al. (2000) J Biol Chem 275, 24977-83.
  6. Bertrand, J. et al. (2010) Neuroscience 168, 323-34.
  7. Yamamoto, H. et al. (2005) J Neurochem 94, 1438-47.
  8. Goedert, M. et al. (1993) Proc Natl Acad Sci U S A 90, 5066-70.

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 3
    #96628

    Phospho-Tau Family Antibody Sampler Kit

    Phospho-Tau Family Antibody Sampler Kit: Image 1 Expand Image
    Simple Western analysis of lysates (0.1 mg/mL) from Mouse Brain Tissue Extracts using Tau (D1M9X) XP® Rabbit #46687. The virtual lane view (left) shows the target band (as indicated) and a band corresponding to Tau (as indicated) at 1:50 and 1:250 dilutions of primary antibody. The corresponding electropherogram view (right) plots chemiluminescence by molecular weight along the capillary at 1:50 (blue line) and 1:250 (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.
    Phospho-Tau Family Antibody Sampler Kit: Image 2 Expand Image
    Confocal immunofluorescent analysis of fixed frozen mouse cerebellum using Tau (D1M9X) XP® Rabbit mAb (green), TMEM119 (E4B9S) Mouse mAb #98778 (red) and ProLong® Gold Antifade Reagent with DAPI #8961 (blue).
    Phospho-Tau Family Antibody Sampler Kit: Image 3 Expand Image
    Confocal immunofluorescent analysis of fixed frozen mouse hippocampus using Tau (D1M9X) XP® Rabbit mAb (green), TMEM119 (E4B9S) Mouse mAb #98778 (red) and ProLong® Gold Antifade Reagent with DAPI #8961 (blue).
    Phospho-Tau Family Antibody Sampler Kit: Image 4 Expand Image
    Western blot analysis of normal mouse brain and Tau KO (-/-) mouse brain with Phospho-Tau (Ser416) (D7U2P) Rabbit mAb (upper) and β-Actin (D6A8) Rabbit mAb #8457 (lower). Tau-KO mouse brain tissue was kindly provided by Dr. Dominic Walsh at Brigham and Women's Hospital and Harvard Medical School.
    Phospho-Tau Family Antibody Sampler Kit: Image 5 Expand Image
    Western blot analysis of normal mouse brain and Tau KO (-/-) mouse brain with Phospho-Tau (Ser404) (D2Z4G) Rabbit mAb (upper) and β-Actin (D6A8) Rabbit mAb #8457 (lower). Tau-KO mouse brain tissue was kindly provided by Dr. Dominic Walsh at Brigham and Women's Hospital and Harvard Medical School.
    Phospho-Tau Family Antibody Sampler Kit: Image 6 Expand Image
    Western blot analysis of normal mouse brain and Tau KO (-/-) mouse brain with Phospho-Tau (Ser404) (D2Z4G) Rabbit mAb (upper) and β-Actin (D6A8) Rabbit mAb #8457 (lower). Tau-KO mouse brain tissue was kindly provided by Dr. Dominic Walsh at Brigham and Women's Hospital and Harvard Medical School.
    Phospho-Tau Family Antibody Sampler Kit: Image 7 Expand Image
    Western blot analysis of normal mouse brain and Tau KO (-/-) mouse brain with Phospho-Tau (Ser202) (D4H7E) Rabbit mAb (upper) and β-Actin (D6A8) Rabbit mAb #8457 (lower). Tau-KO mouse brain tissue was kindly provided by Dr. Dominic Walsh at Brigham and Women's Hospital and Harvard Medical School.
    Phospho-Tau Family Antibody Sampler Kit: Image 8 Expand Image
    Western blot analysis of normal mouse brain and Tau KO (-/-) mouse brain with Tau (D1M9X) XP® Rabbit mAb (upper) and β-Actin (D6A8) Rabbit mAb #8457 (lower). Tau-KO mouse brain tissue was kindly provided by Dr. Dominic Walsh at Brigham and Women's Hospital and Harvard Medical School.
    Phospho-Tau Family Antibody Sampler Kit: Image 9 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.
    Phospho-Tau Family Antibody Sampler Kit: Image 10 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.
    Phospho-Tau Family Antibody Sampler Kit: Image 11 Expand Image
    Western blot analysis of extracts from mouse brain and CAD cells, using Phospho-Tau (Ser396) (PHF13) Mouse mAb. The phospho-specificity of the antibody was verified by peptide blocking using no peptide, phospho-peptide or nonphospho-peptide.
    Phospho-Tau Family Antibody Sampler Kit: Image 12 Expand Image
    Western blot analysis of mouse brain lysate, untreated (-) or λ phosphatase-treated (+), using Phospho-Tau (Ser416) (D7U2P) Rabbit mAb (upper) and β-Actin (D6A8) Rabbit mAb #8457 (lower).
    Phospho-Tau Family Antibody Sampler Kit: Image 13 Expand Image
    Western blot analysis of extracts from human cortex (lane 1), neonatal mouse brain, untreated (lane 2) or phosphatase-treated (lane 3), and fetal rat brain (lane 4), using Phospho-Tau (Ser404) (D2Z4G) Rabbit mAb (upper), Tau (Tau46) Mouse mAb #4019 (middle) and β-Actin (D6A8) Rabbit mAb #8457 (lower).
    Phospho-Tau Family Antibody Sampler Kit: Image 14 Expand Image
    Western blot analysis of extracts from human cortex (lane 1), untreated neonatal mouse brain (lane 2), phosphatase-treated neonatal mouse brain (lane 3), and fetal rat brain (lane 4), using Phospho-Tau (Ser404) (D2Z4G) Rabbit mAb (upper), Tau (Tau46) Mouse mAb #4019 (middle), and β-Actin (D6A8) Rabbit mAb #8457 (lower).
    Phospho-Tau Family Antibody Sampler Kit: Image 15 Expand Image
    Western blot analysis of extracts from mouse brain, untreated (-) or λ-phosphatase-treated (+), using Phospho-Tau (Ser202) (D4H7E) Rabbit mAb (upper) and β-Tubulin (D2N5G) Rabbit mAb #15115 (lower).
    Phospho-Tau Family Antibody Sampler Kit: Image 16 Expand Image
    Western blot analysis of extracts from various cell lines and tissues using Tau (D1M9X) XP® Rabbit mAb (upper) and β-Actin (D6A8) Rabbit mAb #8457 (lower).
    Phospho-Tau Family Antibody Sampler Kit: Image 17 Expand Image
    Western blot analysis of extracts from mouse and rat brain using Phospho-Tau (Ser416) (D7U2P) Rabbit mAb. The phospho-specificity of Phospho-Tau (Ser416) (D7U2P) Rabbit mAb was verified by peptide blocking using a phosphopeptide or non-phosphopeptide targeting residue Ser416.
    Phospho-Tau Family Antibody Sampler Kit: Image 18 Expand Image
    Western blot analysis of extracts from MEF cells (lane 1) and mouse brain (lane 2) using Phospho-Tau (Ser404) (D2Z4G) Rabbit mAb (upper), Tau (Tau46) Mouse mAb #4019 (middle) and β-Actin (D6A8) Rabbit mAb #8457 (lower).
    Phospho-Tau Family Antibody Sampler Kit: Image 19 Expand Image
    Immunohistochemical analysis of paraffin-embedded human Alzheimer's brain using Phospho-Tau (Ser404) (D2Z4G) Rabbit mAb.
    Phospho-Tau Family Antibody Sampler Kit: Image 20 Expand Image
    Western blot analysis of extracts from neonatal and adult mouse brain using Phospho-Tau (Ser202) (D4H7E) Rabbit mAb. The phospho-specificity of the antibody was verified by blocking with a phospho- or nonphosphopeptide.
    Phospho-Tau Family Antibody Sampler Kit: Image 21 Expand Image
    Immunohistochemical analysis of paraffin-embedded human Alzheimer's brain using Tau (D1M9X) XP® Rabbit mAb in the presence of control peptide (left) or antigen-specific peptide (right).
    Phospho-Tau Family Antibody Sampler Kit: Image 22 Expand Image
    Immunohistochemical analysis of paraffin-embedded human breast carcinoma using Phospho-Tau (Ser416) (D7U2P) Rabbit mAb.
    Phospho-Tau Family Antibody Sampler Kit: Image 23 Expand Image
    Confocal immunofluorescent analysis of Tg2576 mouse brain, untreated (left) or Lambda Protein Phosphatase-treated (right), using Phospho-Tau (Ser404) (D2Z4G) Rabbit mAb (red) and GFAP (GA5) Mouse mAb #3670 (green). Blue pseudocolor = DRAQ5® #4084 (fluorescent DNA dye).
    Phospho-Tau Family Antibody Sampler Kit: Image 24 Expand Image
    Immunohistochemical analysis of paraffin-embedded mouse colon, untreated (left) or λ phosphatase-treated (right), using Phospho-Tau (Ser404) (D2Z4G) Rabbit mAb.
    Phospho-Tau Family Antibody Sampler Kit: Image 25 Expand Image
    Immunohistochemical analysis of paraffin-embedded human normal appendix using Tau (D1M9X) XP® Rabbit mAb.
    Phospho-Tau Family Antibody Sampler Kit: Image 26 Expand Image
    Immunohistochemical analysis of paraffin-embedded mouse brain, untreated (left) or λ phosphatase-treated (right), using Phospho-Tau (Ser416) (D7U2P) Rabbit mAb.
    Phospho-Tau Family Antibody Sampler Kit: Image 27 Expand Image
    Confocal immunofluorescent analysis of mouse primary neurons using Phospho-Tau (Ser404) (D2Z4G) Rabbit mAb (green). Blue pseudocolor = Hoescht 33342 #4082 (fluorescent DNA dye).
    Phospho-Tau Family Antibody Sampler Kit: Image 28 Expand Image
    Immunohistochemical analysis of paraffin-embedded T-47D cell pellets (left, positive) and MDA-MB-231 cell pellets (right, negative) using Phospho-Tau (Ser404) (D2Z4G) Rabbit mAb.
    Phospho-Tau Family Antibody Sampler Kit: Image 29 Expand Image
    Immunohistochemical analysis of paraffin-embedded T-47D cell pellet (left, positive) or MDA-MB-231 cell pellet (right, negative) using Tau (D1M9X) XP® Rabbit mAb.
    Phospho-Tau Family Antibody Sampler Kit: Image 30 Expand Image
    Immunohistochemical analysis of paraffin-embedded mouse brain using Phospho-Tau (Ser404) (D2Z4G) Rabbit mAb in the presence of non-phosphorylated tau peptide (left) and antigen-specific phospho-tau (Ser404) peptide (right).
    Phospho-Tau Family Antibody Sampler Kit: Image 31 Expand Image
    Immunohistochemical analysis of paraffin-embedded mouse lung using Tau (D1M9X) XP® Rabbit mAb.
    Phospho-Tau Family Antibody Sampler Kit: Image 32 Expand Image
    Confocal immunofluorescent analysis of fixed frozen mouse striatum using Tau (D1M9X) XP® Rabbit mAb (green), TMEM119 (E4B9S) Mouse mAb #98778 (red) and ProLong® Gold Antifade Reagent with DAPI #8961 (blue).
    Phospho-Tau Family Antibody Sampler Kit: Image 33 Expand Image
    Confocal immunofluorescent analysis of T-47D (positive, left) or MDA-MB-231 (negative, right) cells using Tau (D1M9X) XP® Rabbit mAb (green). Blue pseudocolor = DRAQ5® #4084 (fluorescent DNA dye).