beta Amyloid [Pyro Glu3] Antibody

Immunocytochemistry/Immunofluorescence: beta Amyloid [Pyro Glu3] Antibody [NBP1-44048] - Laser scanning confocal microscopy was carried out using human tissue sections from frontal cortex. Figure A shows an example section that was immunostained for the presence of pyrE3Abeta (green). Figure B shows that the immunogen peptide completely blocked the NBP1-44048 antibody.

Western Blot: beta Amyloid [Pyro Glu3] Antibody [NBP1-44048] - Lane 1-10: beta Amyloid pyro E3 peptide. Load: 1 ug per lane 1-2, 0.2 ug per lane 3-4, 0.05 ug per lane 5-6, 0.01 ug per lane 7-8, 0.005 ug per lane 9-10. Primary antibody: Beta Amyloid pyro E3 antibody at 0.5 ug/mL for overnight at 4C. Secondary antibody: IRDye800 rabbit secondary antibody at 1:10,000 for 45 min at RT. Block: 5% BLOTTO overnight at 4C. Predicted/Observed size: 86.9 kDa for beta Amyloid pyro E3 peptide. Cross reactivity with Beta Amyloid 3 peptide is >1000 fold below pyrE3 reactivity and contains no reactivity towards the Beta Amyloid 42 A beta peptide (data not shown).

Immunocytochemistry/Immunofluorescence: beta Amyloid [Pyro Glu3] Antibody [NBP1-44048] - Tissue: human brain section. Fixation: 0.5% PFA. Antigen retrieval: not required. Primary antibody: beta Amyloid pyro E3 antibody at 5 ug/mL for 1 h at RT. Secondary antibody: Rabbit secondary antibody at 1:10,000 for 45 min at RT. Localization: beta Amyloid pyro E3 is nuclear and cytoplasmic. Staining: Top: Beta Amyloid pyro E3 as as green fluorescent signal, Beta Amyloid 3 as yellow signal; and Bottom: Beta Amyloid 3 as red signal with co-incubation of Beta Amyloid pyro E3 peptide.

Western Blot: beta Amyloid [Pyro Glu3] Antibody [NBP1-44048] - Lane 1: beta Amyloid 3 peptide Lane 2: beta Amyloid pyro E3 peptide. Load: 1 ug per lane. Primary antibody: Beta Amyloid pyro E3 antibody at 2ug/mL for overnight at 4C. Secondary antibody: IRDye800 rabbit secondary antibody at 1:10,000 for 45 min at RT. Block: 5% BLOTTO overnight at 4C. Predicted/Observed size: 86.9 kDa for beta Amyloid pyro E3 peptide.

• Reactivity: Hu, Mu, Ca, Dr
• Applications: WB, ELISA, IA, ICC/IF, IHC, IHC-Fr, IHC-P
• Clonality: Polyclonal
• Host: Rabbit
• Conjugate: Unconjugated

beta Amyloid [Pyro Glu3] Antibody Summary

• Immunogen: beta Amyloid [Pyro Glu3] Antibody was prepared from whole rabbit serum produced by repeated immunizations with a synthetic peptide corresponding to the N-terminus 3-pyro E start point of human beta Amyloid. (Uniprot: P05067)
• Modification: Pyro Glu3
• Specificity: This antibody contains no reactivity towards the 1-42 ABeta peptide.
• Isotype: IgG
• Clonality: Polyclonal
• Host: Rabbit
• Gene: APP
• Purity: Immunogen affinity purified

Applications/Dilutions

• Dilutions:
  • ELISA 1:20000-1:60000
  • Immunoassay
  • Immunocytochemistry/Immunofluorescence 1:100-1:500
  • Immunohistochemistry 1:100-1:500
  • Immunohistochemistry-Frozen
  • Immunohistochemistry-Paraffin 1:500-1:1000
  • Western Blot 1 ug/ml
• Application Notes: This product has been tested in dot blot, ELISA, Western Blot, and Immunostaining. Specific conditions for reactivity should be optimized by the end user. Expect a band approximately ~86.9kDa corresponding to the appropriate cell lysate or extract.
  
  Use in Immunohistochemistry-Frozen reported in scientific literature (PMID: 27776266).
  
  Use in Immunoassay reported in scientific literature (PMID: 26208119).

Reactivity Notes

A BLAST analysis was used to suggest cross-reactivity with Primate, Bovine and Sheep based on 100% sequence homology.
Mouse reactivity reported in scientific literature (PMID: 31434879).

Packaging, Storage & Formulations

• Storage: Store at -20C short term. Aliquot and store at -80C long term. Avoid freeze-thaw cycles.
• Buffer: 0.02 M Potassium Phosphate, 0.15 M Sodium Chloride, pH 7.2
• Preservative: 0.01% Sodium Azide
• Purity: Immunogen affinity purified

Notes

This antibody was affinity purified from monospecific antiserum by immunoaffinity chromatography

Store vial at -20C prior to opening. Aliquot contents and freeze at -20C or below for extended storage. Avoid cycles of freezing and thawing. Centrifuge product if not completely clear after standing at room temperature. This product is stable for several weeks at 4C as an undiluted liquid. Dilute only prior to immediate use.

Alternate Names for beta Amyloid [Pyro Glu3] Antibody

• Abeta
• Amyloid beta
• beta-Amyloid Peptide

Background

Beta amyloid, also known as Abeta, is a peptide that varies in length from 39 to 43 amino acids and is processed from the transmembrane protein amyloid precursor protein (APP) by beta-secretase (BACE-1) and gamma-secretase. Beta amyloid [1-40], beta amyloid [1-42], and beta amyloid [1-43] peptides result from cleavage of APP after residues 40, 42, and 43, respectively. The aggregation of beta amyloid monomers produces insoluble oligomers and protofibrils. The longer "stickier" forms of beta amyloid, particularly ABeta42, are known as the main component of amyloid plaques, which are deposits (senile plaques or neuritic plaques) found in the brains of patients with Alzheimer's disease (AD) (1,2). Research indicates that intraneuronal beta amyloid accumulation may be an important proximal neurotoxic event in the pathogenesis of Alzheimer's disease. Abeta deposits have also been reported in the hearts of AD patients (3).

Pyroglutamate amyloid beta peptides (pGlu-Abeta) are N-terminal truncations in which the N-terminal glutamate is cyclized to pyroglutamate resulting in pGlu-Abeta (3-40/42 and 11-40/42) (4). This pyrE modification may have a greater propensity to aggregate under physiological conditions and has been implicated as the molecular species responsible for seeding larger oligomers of amyloid beta.

References

1. Chen GF, Xu TH, Yan Y, Zhou YR, Jiang Y, Melcher K, Xu HE. (2017) Amyloid beta: structure, biology and structure-based therapeutic development. Acta Pharmacol Sin. 38(9):1205-1235. PMID: 28713158

2. De-Paula VJ1, Radanovic M, Diniz BS, Forlenza OV. (2012) Alzheimer's disease. Subcell Biochem. 65:329-52. PMID: 23225010

3. Schaich CL, Maurer MS, Nadkarni NK. (2019) Amyloidosis of the Brain and Heart: Two Sides of the Same Coin? JACC Heart Fail. 7(2):129-131. PMID: 30704604

4. He W, Barrow CJ. (1999) The A beta 3-pyroglutamyl and 11-pyroglutamyl peptides found in senile plaque have greater beta-sheet forming and aggregation propensities in vitro than full-length A beta. Biochemistry. 38(33):10871-7. PMID: 10451383

Limitations

This product is for research use only and is not approved for use in humans or in clinical diagnosis. Primary Antibodies are guaranteed for 1 year from date of receipt.

Publications for beta Amyloid Antibody (NBP1-44048)(5)

Publications using NBP1-44048

• Spangenberg E, Severson PL, Hohsfield LA et al. Sustained microglial depletion with CSF1R inhibitor impairs parenchymal plaque development in an Alzheimer's disease model Nat Commun 2019-08-21 [PMID: 31434879] (IHC, Mouse)
• Davis PR, Giannini G, Rudolph K et al. AB vaccination in combination with behavioral enrichment in aged beagles: effects on cognition, AB, and microhemorrhages. Neurobiol Aging. [PMID: 27776266] (IHC-Fr, Canine)
• Jonson M, Pokrzywa M, Starkenberg A et al. Systematic Ab Analysis in Drosophila Reveals High Toxicity for the 1-42, 3-42 and 11-42 Peptides, and Emphasizes N- and C-Terminal Residues PLoS ONE. 2015-07-25 [PMID: 26208119] (IA, Drosophila)
• Schilling S, Lauber T, Schaupp M, Manhart S, Scheel E, Bohm G, Demuth HU. On the seeding and oligomerization of pGlu-amyloid peptides (in vitro). Biochemistry;45(41):12393-9. 2006-10-17 [PMID: 17029395]
• He W, Barrow CJ. The A beta 3-pyroglutamyl and 11-pyroglutamyl peptides found in senile plaque have greater beta-sheet forming and aggregation propensities in vitro than full-length A beta. Biochemistry;38(33):10871-7. 1999-08-17 [PMID: 10451383]

Bioinformatics

• Gene Symbol: APP
• Entrez:
  • 351(Human)
• Uniprot:
  • P05067()
  • P05067()
  • P05067()
  • P05067()
  • P05067()