Revision 2
#74113
Store at -20C
Pathological Hallmarks of Alzheimer's Disease Antibody Sampler Kit
1 Kit
(9 x 20 microliters)
877-616-CELL (2355)
877-678-TECH (8324)
3 Trask Lane | Danvers | Massachusetts | 01923 | USA
For Research Use Only. Not for Use in Diagnostic Procedures.
| Product Includes | Product # | Quantity | Mol. Wt | Isotype/Source |
|---|---|---|---|---|
| β-Amyloid (D54D2) XP® Rabbit mAb | 8243 | 20 µl | 5 kDa | Rabbit IgG |
| β-Amyloid (1-42) (D9A3A) Rabbit mAb | 14974 | 20 µl | 4 kDa | Rabbit IgG |
| β-Amyloid (1-40) (D8Q7I) Rabbit mAb | 12990 | 20 µl | 4 kDa | Rabbit IgG |
| β-Amyloid (1-43) (E8C2D) Rabbit mAb | 32098 | 20 µl | 6 kDa | Rabbit IgG |
| β-Amyloid (pE3 Peptide) (D5N5H) Rabbit mAb | 14975 | 20 µl | 4 kDa | Rabbit IgG |
| Tau (D1M9X) XP® Rabbit mAb | 46687 | 20 µl | 50-80 kDa | Rabbit IgG |
| Phospho-Tau (Thr205) (E7D3E) Rabbit mAb | 49561 | 20 µl | 50-80 kDa | Rabbit IgG |
| Phospho-Tau (Ser404) (D2Z4G) Rabbit mAb | 20194 | 20 µl | 50-80 kDa | Rabbit IgG |
| Phospho-Tau (Thr181) (D9F4G) Rabbit mAb | 12885 | 20 µl | 50-80 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 Pathological Hallmarks of Alzheimer's Disease Antibody Sampler Kit provides an economical means of detecting the activation of Tau and APP family members using phospho-specific, and control antibodies for both proteins. The kit includes enough antibodies to perform two western blot experiments with each primary antibody.
Storage
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, 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; these tangles are bundles of paired helical filaments 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). The cerebrospinal fluid concentration of tau phosphorylated at Thr181 has been proposed to be a biomarker for the study of neurodegenerative disorders (4).
Amyloid β (Aβ) precursor protein (APP) is a 100-140 kDa transmembrane glycoprotein that exists as several isoforms (4). The amino acid sequence of APP contains the amyloid domain, which can be released by a two-step proteolytic cleavage (4). The extracellular deposition and accumulation of the released Aβ fragments form the main components of amyloid plaques in Alzheimer's disease (4). APP can be phosphorylated at several sites, which may affect the proteolytic processing and secretion of this protein (5-8). Aβ-43 has been suggested as a biomarker in early onset of Alzheimer's disease, where patients have lower levels of Aβ-43 in cerebrospinal fluid (8-10). Several studies have shown that Aβ toxicity of Aβ-43 is as high as Aβ-42 or Aβ-40 in different models of Alzheimer's disease, including mouse models and human disease (10).
Background References
- Johnson, G.V. and Stoothoff, W.H. (2004) J Cell Sci 117, 5721-9.
- Hanger, D.P. et al. (1998) J Neurochem 71, 2465-76.
- Bramblett, G.T. et al. (1993) Neuron 10, 1089-99.
- Mitchell, A.J. (2009) J Neurol Neurosurg Psychiatry 80, 966-75.
- Selkoe, D.J. (1996) J Biol Chem 271, 18295-8.
- Caporaso, G.L. et al. (1992) Proc Natl Acad Sci U S A 89, 3055-9.
- Hung, A.Y. and Selkoe, D.J. (1994) EMBO J 13, 534-42.
- Suzuki, T. et al. (1994) EMBO J 13, 1114-22.
- Ando, K. et al. (1999) J Neurosci 19, 4421-7.
- Iijima, K. et al. (2000) J Neurochem 75, 1085-91.
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
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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 2
Western blot analysis of normal mouse brain and Tau KO (-/-) mouse brain with Phospho-Tau (Thr181) (D9F4G) 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.
Western blot analysis of brain extracts from 13-month old wild-type and TG2576 mice using β-Amyloid (1-40) (D8Q7I) Rabbit mAb (upper) and β-Actin (D6A8) Rabbit mAb #8457 (lower).
Western blot analysis of human Aβ-37, Aβ-38, Aβ-39, Aβ-40, Aβ-42, and Aβ-43 peptides (2.5 ng) using β-Amyloid (1-42) (D9A3A) Rabbit mAb (upper) and β-Amyloid (D54D2) XP® Rabbit mAb #8243 (lower).</p>
Revision 2
β-Amyloid (pE3 Peptide) (D5N5H) Rabbit mAb specificity is demonstrated using peptide dot blot. β-Amyloid (pE3 Peptide) (D5N5H) Rabbit mAb binds to pre-coated β-amyloid (pE3) peptide but not to β-amyloid (E3) peptide or scrambled β-amyloid (pE3) peptide.
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.
Western blot analysis with the indicated amounts of human β-amyloid (1-43) protein using β-Amyloid (1-43) (E8C2D) Rabbit mAb.
Revision 2
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.
Western blot analysis of normal mouse brain and Tau KO (-/-) mouse brain with Phospho-Tau (Thr205) (E7D3E) 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.<
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.
Revision 2
Western blot analysis of human Aβ-42, Aβ-40, Aβ-39, Aβ-38, and Aβ-37 peptides (5 ng) using β-Amyloid (D54D2) XP® Rabbit mAb.
Western blot analysis of extracts from mouse and rat brain using Phospho-Tau (Thr181) (D9F4G) Rabbit mAb. The phospho-specificity of Phospho-Tau (Thr181) (D9F4G) Rabbit mAb was verified by peptide blocking using a phosphopeptide or non-phosphopeptide targeting residue Thr181.
Western blot analysis of human Aβ-37, Aβ-38, Aβ-39, Aβ-40, Aβ-42 and Aβ-43 peptides (10 ng) using β-Amyloid (1-40) (D8Q7I) Rabbit mAb (upper) and β-Amyloid (D54D2) XP® Rabbit mAb #8243 (lower).
Revision 2
Western blot analysis of brain extracts from 13-month old wild-type and Tg2576 mouse model of Alzheimer's brain using β-Amyloid (1-42) (D9A3A) Rabbit mAb (upper) and β-Actin (D6A8) Rabbit mAb #8457 (lower).
Western blot analysis of the indicated amounts of human Aβ (pE3-42) and Aβ (1-42) peptides using β-Amyloid (pE3 Peptide) (D5N5H) Rabbit mAb (upper) and β-Amyloid (1-42) (D3E10) Rabbit mAb #12843 (lower).
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).
Revision 2
Western blot analysis of human Aβ-37, Aβ-38, Aβ-39, Aβ-40, Aβ-42 and Aβ-43 peptides (10 ng) using β-Amyloid (1-43) (E8C2D) Rabbit mAb. Note the slight cross-reactivity with Aβ-42.
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).
Western blot analysis of extracts from mouse brain, untreated (-) or phosphatase-treated (+), and rat brain using Phospho-Tau (Thr205) (E7D3E) Rabbit mAb (upper) and Tau (D1M9X) Rabbit mAb #46687 (lower).
Revision 2
Western blot analysis of the indicated amounts of human Aβ-42 (left) and Aβ-40 (right) peptides using β-Amyloid (D54D2) XP® Rabbit mAb.
Confocal immunofluorescent analysis of hippocampus from wild-type (left) and TG2576 (right) mice using β-Amyloid (1-40) (D8Q7I) Rabbit mAb (green) and β3-Tubulin (TU-20) Mouse mAb #4466 (red). Blue pseudocolor = DRAQ5® #4084 (fluorescent DNA dye).
Immunohistochemical analysis of paraffin-embedded human Alzheimer's brain using β-Amyloid (pE3 Peptide) (D5N5H) Rabbit mAb.
Revision 2
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).
Confocal immunofluorescent analysis of brain from an amyloid mouse model of Alzheimer's disease using β-Amyloid (1-43) (E8C2D) Rabbit mAb (green). After blocking free secondary antibody binding sites with Rabbit (DA1E) mAb IgG XP® Isotype Control #3900, the tissue was then labeled using GFAP (GA5) Mouse mAb (Alexa Fluor® 555 Conjugate) #3656 (red pseudocolor) and β-Amyloid (D54D2) XP® Rabbit mAb (Alexa Fluor® 647 Conjugate) #42284 (yellow pseudocolor). Samples were mounted in ProLong® Gold Antifade Reagent with DAPI #8961 (blue).
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).
Revision 2
Immunoprecipitation of Phospho-Tau (Thr205) from mouse brain lysates. Lane 1 mouse brain lysate immunoprecipitation, lane 2 is Rabbit (DA1E) mAb IgG XP® Isotype Control #3900, lanes 3, 4, and 5 represent three dilutions of the immunoprecipitation of mouse brain lysates at 1:50, 1:100, 1:200 respectively, and lane 6 is mouse brain lysate. Western blot analysis was performed using Phospho-Tau (Thr205) (E7D3E) Rabbit mAb.
Western blot analysis of human Aβ-42 peptide (1 ng) and human cerebrospinal fluid (CSF) of an AD patient using β-Amyloid (D54D2) XP® Rabbit mAb.
Immunohistochemical analysis of paraffin-embedded human Alzheimer's brain using Phospho-Tau (Thr181) (D9F4G) Rabbit mAb.
Revision 2
Confocal immunofluorescent analysis of wild-type (left) and Tg2576 mouse model of Alzheimer's brain (right) using β-Amyloid (1-42) (D9A3A) Rabbit mAb (green) and β3-Tubulin (TU-20) Mouse mAb #4466 (red). Blue pseudocolor = DRAQ5® #4084 (fluorescent DNA dye).
Immunohistochemical analysis of paraffin-embedded Tg2576 mouse model of Alzheimer's brain using β-Amyloid (pE3 Peptide) (D5N5H) Rabbit mAb.
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).
Revision 2
Immunohistochemical analysis of paraffin-embedded human normal appendix using Tau (D1M9X) XP® Rabbit mAb.
Immunohistochemical analysis of paraffin-embedded human Alzheimer's disease brain using Phospho-Tau (Thr205) (E7D3E) Rabbit mAb in the presence of non-phospho-Tau (Thr205) peptide (left) or phospho-Tau (Thr205) peptide (right).
Immunohistochemical analysis of paraffin-embedded human breast carcinoma using Phospho-Tau (Thr181) (D9F4G) Rabbit mAb.
Revision 2
Confocal immunofluorescent analysis of wild-type (left) and Tg2576 mouse model of Alzheimer's brain (right) using β-Amyloid (pE3 Peptide) (D5N5H) Rabbit mAb (green) and β3-Tubulin (TU-20) Mouse mAb #4466 (red). Blue pseudocolor = DRAQ5® #4084 (fluorescent DNA dye).
Confocal immunofluorescent analysis of mouse primary neurons using Phospho-Tau (Ser404) (D2Z4G) Rabbit mAb (green). Blue pseudocolor = Hoescht 33342 #4082 (fluorescent DNA dye).
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.
Revision 2
Immunohistochemical analysis of paraffin-embedded mouse lung using Tau (D1M9X) XP® Rabbit mAb.
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).
Confocal immunofluorescent analysis of mouse medulla oblangata using Phospho-Tau (Thr205) (E7D3E) Rabbit mAb (green). Blue = DAPI #4083 (fluorescent DNA dye).
Revision 2
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).
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.
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).
Revision 2
Immunohistochemical analysis of paraffin-embedded mouse brain using Phospho-Tau (Thr205) (E7D3E) Rabbit mAb.
Confocal immunofluorescent analysis of mouse subicular cortex from an amyloid mouse model of Alzheimer's Disease using β-Amyloid (D54D2) XP® Rabbit mAb #8243 (green) and GFAP (GA5) Mouse mAb #3670 (red). Samples were mounted in ProLong® Gold Antifade Reagent with DAPI #8961 (blue).
Immunohistochemical analysis of paraffin-embedded mouse brain using Phospho-Tau (Thr181) (D9F4G) Rabbit mAb.
Revision 2
Immunohistochemical analysis of paraffin-embedded mouse small intestine using Phospho-Tau (Thr205) (E7D3E) Rabbit mAb.
Immunohistochemical analysis of paraffin-embedded mouse colon, control (left) or λ phosphatase-treated (right), using Phospho-Tau (Thr181) (D9F4G) Rabbit mAb.
Confocal immunofluorescent analysis of mouse brainstem using Phospho-Tau (Thr181) (D9F4G) Rabbit mAb #12885 (green) and S6 Ribosomal Protein (54D2) Mouse mAb #2317 (red). Antibody was pre-incubated with non-phospho-Tau (Thr181) peptide (left), a phospho-Tau (Thr181) peptide (middle), or without peptide (right) to confirm phospho-specificity. Samples were mounted in ProLong® Gold Antifade Reagent with DAPI #8961 (blue).
Revision 2
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).
Confocal immunofluorescent analysis of dentate gyrus in wild-type mouse brain using Phospho-Tau (Thr205) (E7D3E) Rabbit mAb (green). Antibody was pre-incubated with a non-phospho-Tau peptide (left), a phospho-Tau (Thr205) peptide (center), or without peptide (right) to confirm phospho-specificity. Samples were mounted in ProLong® Gold Antifade Reagent with DAPI #8961 (blue).
