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  • Immunohistochemical characterization of novel monoclonal antibodies against the N-terminus of amyloid β-peptide.

Immunohistochemical characterization of novel monoclonal antibodies against the N-terminus of amyloid β-peptide.

Amyloid : the international journal of experimental and clinical investigation : the official journal of the International Society of Amyloidosis (2013-07-09)
Nicolaas A Verwey, Jeroen J M Hoozemans, Carsten Korth, Marloes R van Royen, Ingrid Prikulis, Dorine Wouters, Harry A M Twaalfhoven, Elise S van Haastert, Dale Schenk, Philip Scheltens, Annemieke J M Rozemuller, Marinus A Blankenstein, Robert Veerhuis
ABSTRACT

Abstract Amyloid β-peptide (Aβ) is a key molecule in Alzheimer's disease (AD). Reliable immunohistochemical (IHC) methods to detect Aβ and Aβ-associated factors (AAF) in brain specimens are needed to determine their role in AD pathophysiology. Formic acid (FA) pre-treatment, which is generally used to enable efficient detection of Aβ with IHC, induces structural modifications within the Aβ, as well as in AAF. Consequently, interpretation of double IHC stainings becomes difficult. Therefore, serial stainings of two newly produced monoclonal antibodies (mAbs) VU-17 and IC16 and two other mAbs (6E10 and 3D6) were performed with four different pre-treatments (no pre-treatment, Tris/EDTA, citrate and FA) and additionally six IHC characteristics were scored: diffuse/compact/classic plaques, arteries with cerebral Aβ angiopathy, dyshoric angiopathy, capillaries with dyshoric angiopathy. Subsequently, these stainings were compared with IHC procedures, which are frequently used in a diagnostic setting, employing mAbs 4G8 and 6F/3D with FA pre-treatment. IHC Aβ patterns obtained with VU-17 and, IC16 and 3D6 without the use of FA pre-treatment were comparable to those obtained with 4G8 and 6F/3D upon FA pre-treatment. Omission of FA pre-treatment gives the advantage to allow double IHC stainings, detecting both Aβ and AAF that otherwise would have been structural modificated upon FA pre-treatment.

MATERIALS
Product Number
Brand
Product Description

Supelco
Calcium formate, Standard for quantitative NMR, TraceCERT®, Manufactured by: Sigma-Aldrich Production GmbH, Switzerland
Sigma-Aldrich
Ammonium formate, BioUltra, ≥99.0% (calc. based on dry substance, NT)
Supelco
Ammonium formate, eluent additive for LC-MS, LiChropur, ≥99.0%
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Thallium(I) formate, 97%
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Ammonium formate, ≥99.995% trace metals basis
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Cesium formate, 98%
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Calcium formate, BioUltra, ≥99.0% (T)
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Ammonium formate, reagent grade, 97%
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Sodium formate-13C, 99 atom % 13C
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Sodium formate, 99.998% trace metals basis
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Formic acid, ≥95%, FCC, FG
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Formic acid solution, BioUltra, 1.0 M in H2O
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Ammonium formate solution, BioUltra, 10 M in H2O
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Potassium formate, ReagentPlus®, 99%
Sigma-Aldrich
Potassium formate, BioUltra, ≥99.0% (NT)
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Sodium formate, BioUltra, ≥99.0% (NT)
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Sodium formate, reagent grade, 97%
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Formic acid, ACS reagent, ≥96%
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Formic acid, puriss., meets analytical specifications of DAC, FCC, 98.0-100%
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Sodium formate, ACS reagent, ≥99.0%
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Formic acid, reagent grade, ≥95%
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Formic acid, puriss. p.a., ACS reagent, reag. Ph. Eur., ≥98%
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Formic acid, ACS reagent, ≥88%