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74113
Pathological Hallmarks of Alzheimer's Disease Antibody Sampler Kit
Primary Antibodies

Pathological Hallmarks of Alzheimer's Disease Antibody Sampler Kit #74113

Western Blotting Image 1

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.

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Western Blotting Image 2

Western blot analysis of human Aβ-37, Aβ-38, Aβ-39, Aβ-40, and Aβ-42 peptides (2.5 ng) using β-Amyloid (1-42 Preferred) (D9A3A) Rabbit mAb (upper) and β-Amyloid (D54D2) XP® Rabbit mAb #8243 (lower).

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Western Blotting Image 3

Western blot analysis of brain extracts from 13-month old wild-type and TG2576 mice using β-Amyloid (1-40 Specific) (D8Q7I) Rabbit mAb (upper) and β-Actin (D6A8) Rabbit mAb #8457 (lower).

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Western Blotting Image 4

Western blot analysis with the indicated amounts of human β-amyloid (1-43) protein using β-Amyloid (1-43 Preferred) (E8C2D) Rabbit mAb.

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Western Blotting Image 5

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.

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Western Blotting Image 6

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).

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Western Blotting Image 7

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).

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Western Blotting Image 8

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 (IF preferred) (upper), Tau (Tau46) Mouse mAb #4019 (middle) and β-Actin (D6A8) Rabbit mAb #8457 (lower).

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IHC-P (paraffin) Image 9

Immunohistochemical analysis of paraffin-embedded human Alzheimer's brain using Phospho-Tau (Thr181) (D9F4G) Rabbit mAb.

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Western Blotting Image 10

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.

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Western Blotting Image 11

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.

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Western Blotting Image 12

Western blot analysis of the indicated amounts of human Aβ-42 (left) and Aβ-40 (right) peptides using β-Amyloid (D54D2) XP® Rabbit mAb.

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Western Blotting Image 13

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 Preferred) (D9A3A) Rabbit mAb (upper) and β-Actin (D6A8) Rabbit mAb #8457 (lower).

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Western Blotting Image 14

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 Specific) (D8Q7I) Rabbit mAb (upper) and β-Amyloid (D54D2) XP® Rabbit mAb #8243 (lower).

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Western Blotting Image 15

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 Preferred) (E8C2D) Rabbit mAb. Note the slight cross-reactivity with Aβ-42.

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Western Blotting Image 16

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 Specific) (D3E10) Rabbit mAb #12843 (lower).

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IHC-P (paraffin) Image 17

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).

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IP Image 18

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.

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Western Blotting Image 19

Western blot analysis of extracts from MEF cells (lane 1) and mouse brain (lane 2) using Phospho-Tau (Ser404) (D2Z4G) Rabbit mAb (IF preferred) (upper), Tau (Tau46) Mouse mAb #4019 (middle) and β-Actin (D6A8) Rabbit mAb #8457 (lower).

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Western Blotting Image 20

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.

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IF-F Image 21

Confocal immunofluorescent analysis of wild-type (left) and Tg2576 mouse model of Alzheimer's brain (right) using β-Amyloid (1-42 Preferred) (D9A3A) Rabbit mAb (green) and β3-Tubulin (TU-20) Mouse mAb #4466 (red). Blue pseudocolor = DRAQ5® #4084 (fluorescent DNA dye).

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IF-F Image 22

Confocal immunofluorescent analysis of hippocampus from wild-type (left) and TG2576 (right) mice using β-Amyloid (1-40 Specific) (D8Q7I) Rabbit mAb (green) and β3-Tubulin (TU-20) Mouse mAb #4466 (red). Blue pseudocolor = DRAQ5® #4084 (fluorescent DNA dye).

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IF-F Image 23

Confocal immunofluorescent analysis of brain from the 5XFAD mouse model of Alzheimer's disease using β-Amyloid (1-43 Preferred) (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).

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IHC-P (paraffin) Image 24

Immunohistochemical analysis of paraffin-embedded human Alzheimer's brain using β-Amyloid (pE3 Peptide) (D5N5H) Rabbit mAb.

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IHC-P (paraffin) Image 25

Immunohistochemical analysis of paraffin-embedded human normal appendix using Tau (D1M9X) XP® Rabbit mAb.

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IHC-P (paraffin) Image 26

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).

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IF-IC Image 27

Confocal immunofluorescent analysis of mouse primary neurons using Phospho-Tau (Ser404) (D2Z4G) Rabbit mAb (IF preferred) (green). Blue pseudocolor = Hoescht 33342 #4082 (fluorescent DNA dye).

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IHC-P (paraffin) Image 28

Immunohistochemical analysis of paraffin-embedded human breast carcinoma using Phospho-Tau (Thr181) (D9F4G) Rabbit mAb.

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IF-P Image 29

Confocal immunofluorescent analysis of paraffin-embedded human Alzheimer's brain using β-Amyloid (D54D2) XP® Rabbit mAb (green) and Tau (Tau46) Mouse mAb #4019 (red). Blue pseudocolor = DRAQ5® #4084 (fluorescent DNA dye).

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IHC-P (paraffin) Image 30

Immunohistochemical analysis of paraffin-embedded Tg2576 mouse model of Alzheimer's brain using β-Amyloid (pE3 Peptide) (D5N5H) Rabbit mAb.

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IHC-P (paraffin) Image 31

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.

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IHC-P (paraffin) Image 32

Immunohistochemical analysis of paraffin-embedded mouse brain using Phospho-Tau (Thr205) (E7D3E) Rabbit mAb.

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IF-F Image 33

Confocal immunofluorescent analysis of Tg2576 mouse brain, untreated (left) or Lambda Protein Phosphatase-treated (right), using Phospho-Tau (Ser404) (D2Z4G) Rabbit mAb (IF preferred) (red) and GFAP (GA5) Mouse mAb #3670 (green). Blue pseudocolor = DRAQ5® #4084 (fluorescent DNA dye).

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IHC-P (paraffin) Image 34

Immunohistochemical analysis of paraffin-embedded mouse brain using Phospho-Tau (Thr181) (D9F4G) Rabbit mAb.

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IF-F Image 35

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).

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IHC-P (paraffin) Image 36

Immunohistochemical analysis of paraffin-embedded mouse lung using Tau (D1M9X) XP® Rabbit mAb.

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IHC-P (paraffin) Image 37

Immunohistochemical analysis of paraffin-embedded mouse small intestine using Phospho-Tau (Thr205) (E7D3E) Rabbit mAb.

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IHC-P (paraffin) Image 38

Immunohistochemical analysis of paraffin-embedded mouse colon, control (left) or λ phosphatase-treated (right), using Phospho-Tau (Thr181) (D9F4G) Rabbit mAb.

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IF-IC Image 39

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).

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IF-F Image 40

Confocal immunofluorescent analysis of mouse medulla oblangata using Phospho-Tau (Thr205) (E7D3E) Rabbit mAb (green). Blue = DAPI #4083 (fluorescent DNA dye).

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IF-F Image 41

Confocal immunofluorescent analysis of mouse striatum (left), hippocampus (middle), or midbrain (right) using Tau (D1M9X) XP® Rabbit mAb (green) and GFAP (GA5) Mouse mAb #3670 (red). Blue pseudocolor = DRAQ5® #4084 (fluorescent DNA dye).

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IF-F Image 42

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).

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Product Includes Quantity Applications Reactivity MW(kDa) Isotype
β-Amyloid (D54D2) XP® Rabbit mAb 8243 20 µl
  • WB
  • IP
  • IF
H 5 Rabbit IgG
β-Amyloid (1-42 Preferred) (D9A3A) Rabbit mAb 14974 20 µl
  • WB
  • IF
H 4 Rabbit IgG
β-Amyloid (1-40 Specific) (D8Q7I) Rabbit mAb 12990 20 µl
  • WB
  • IF
H 4 Rabbit IgG
β-Amyloid (1-43 Preferred) (E8C2D) Rabbit mAb 32098 20 µl
  • WB
  • IF
H 6 Rabbit IgG
β-Amyloid (pE3 Peptide) (D5N5H) Rabbit mAb 14975 20 µl
  • WB
  • IHC
  • IF
H 4 Rabbit IgG
Tau (D1M9X) XP® Rabbit mAb 46687 20 µl
  • WB
  • IHC
  • IF
H M R 50 to 80 Rabbit IgG
Phospho-Tau (Thr205) (E7D3E) Rabbit mAb 49561 20 µl
  • WB
  • IP
  • IHC
  • IF
H M R 50-80 Rabbit IgG
Phospho-Tau (Ser404) (D2Z4G) Rabbit mAb (IF preferred) 20194 20 µl
  • WB
  • IP
  • IF
H M R 50-80 Rabbit IgG
Phospho-Tau (Thr181) (D9F4G) Rabbit mAb 12885 20 µl
  • WB
  • IP
  • IHC
  • IF
H M R 50-80 Rabbit IgG
Anti-rabbit IgG, HRP-linked Antibody 7074 100 µl
  • WB
Goat 

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.

Pathological Hallmarks of Alzheimer's Disease Antibody Sampler Kit includes Tau (D1M9X) XP® Rabbit mAb that recognizes total tau, and antibodies that recognize various phosphorylated tau epitopes found in Alzheimer's disease, including Phospho-Tau (Thr205) (E7D3E), Phospho-Tau (Ser404) (D2Z4G) (IF preferred), and Phospho-Tau (Thr181) (D9F4G). Additionally, this kit includes antibodies that detect various β-Amyloid peptides of the APP human protein, including β-Amyloid (D54D2) XP® Rabbit mAb, which also detects transgenically expressed human APP in mouse. β-Amyloid (pE3 Peptide) (D5N5H) Rabbit mAb recognizes recombinant pE3 peptides but does not cross-react with the non-E3 peptides. β-Amyloid (1-40 Specific) (D8Q7I) Rabbit mAb does not cross-react with other β-amyloid peptides. β-Amyloid (1-43 Preferred) (E8C2D) Rabbit mAb and β-Amyloid (1-42 preferred) (D9A3A) Rabbit mAb both detect transgenically expressed human APP in mouse models.

Tau antibodies include peptides corresponding to residues surrounding Asp430, phosphorylated at Thr205, Ser400/Thr403/Ser404, and Thr181 of human tau. Monoclonal antibodies are produced by immunizing rabbits with synthetic Aβ peptides corresponding to residues near the carboxy terminus of human β-amyloid (1-42), (1-40), (1-43), (pE3) peptide, and several peptides of Aβ, such as Aβ-37, Aβ-38, Aβ-39, Aβ-40, and Aβ-42.

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).

  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. Mitchell, A.J. (2009) J Neurol Neurosurg Psychiatry 80, 966-75.
  5. Selkoe, D.J. (1996) J Biol Chem 271, 18295-8.
  6. Caporaso, G.L. et al. (1992) Proc Natl Acad Sci U S A 89, 3055-9.
  7. Hung, A.Y. and Selkoe, D.J. (1994) EMBO J 13, 534-42.
  8. Suzuki, T. et al. (1994) EMBO J 13, 1114-22.
  9. Ando, K. et al. (1999) J Neurosci 19, 4421-7.
  10. Iijima, K. et al. (2000) J Neurochem 75, 1085-91.
Entrez-Gene Id
351 , 4137
Swiss-Prot Acc.
P05067 , P10636-8
For Research Use Only. Not For Use In Diagnostic Procedures.

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