IDE Antikörper (AA 491-590)

Produktnummer ABIN723680

Produktdetails anti-IDE Antikörper

Target: IDE (Insulin-Degrading Enzyme (IDE))

Bindungsspezifität: AA 491-590

Reaktivität: Human, Maus, Ratte

Wirt: Kaninchen

Klonalität: Polyklonal

Konjugat: Dieser IDE Antikörper ist unkonjugiert

Applikation: Western Blotting (WB), ELISA, Immunohistochemistry (Paraffin-embedded Sections) (IHC (p)), Immunofluorescence (Cultured Cells) (IF (cc)), Immunofluorescence (Paraffin-embedded Sections) (IF (p)), Immunohistochemistry (Frozen Sections) (IHC (fro))

Kreuzreaktivität: Human, Maus, Ratte

Homologie: Cow,Pig,Chicken

Aufreinigung: Purified by Protein A.

Immunogen: KLH conjugated synthetic peptide derived from human IDE

Isotyp: IgG

Anwendungsinformationen

Applikationshinweise: WB 1:300-5000
ELISA 1:500-1000
IHC-P 1:200-400
IHC-F 1:100-500
IF(IHC-P) 1:50-200
IF(IHC-F) 1:50-200
IF(ICC) 1:50-200

Beschränkungen: Nur für Forschungszwecke einsetzbar

Independent Validation (IHC)

Validierung #104435 (Immunohistochemistry)

by: Prof. Merighi, Laboratory of Neurobiology, Department of Veterinary Sciences, University of Turin

No.: #104435

Datum: 15.03.2023

Antigen: IDE

Chargennummer: 9C07M588

Validierte Anwendung: Immunohistochemistry

Positivkontrolle:

Adult mouse liver fixed in 4% paraformaldehyde

Negativkontrolle:

One control slice for each experimental procedure processed omitting the primary antibody; overnight incubation in diluent solution only.

Bewertung:

Passed. The IDE antibody (AA 491-590) ABIN723680 works in IHC-P at 1:100 concentrations with Tyramide amplification.

Validierungsbilder

Staining of IDE positive cells in the adult mouse liver with ABIN723680.

Western blot detection of IDE (110 kDa) in adult mouse brain (1), cerebellum (2), and liver (3) tissue homogenates with ABIN723680.

Protokoll

Primärantikörper: ABIN723680

Sekundärantikörper: poly-HRP conjugated goat anti-rabbit antibody

Full Protocol:

• Perfuse mice with paraformaldehyde 4% in 0.1 M phosphate buffer pH 7.4 and post-fix in the same fixative for an additional 2 h at RT.
• Wash, dehydrate, and embed samples in paraffin wax.
• Wash several times with 0.01 M PBS.
• Cut liver with a microtome into 20 µm sections and mount on glass slides.
• After paraffin removal, incubate sections for 1 h at RT in PBS containing 1% albumin from chicken egg white (Sigma, A5378) and 0.3% Triton-X-100 (BioRad, 161-0407, lot 00583) to block non-specific binding sites.
• Incubate sections with primary rabbit anti-IDE (antibodies-online, ABIN723680, lot 9C07M588) diluted 1:50, 1:100, 1:200, and 1:300 in PBS-BSA-PLL ON at RT in a humid chamber.
• Wash sections 3x 5 min with 0.01 M PBS.
• Incubate sections with secondary poly-HRP conjugated goat anti-rabbit antibody from Alexa Fluor 488 Tyramide SuperBoost Kit, goat anti-rabbit IgG (Thermo Fisher Scientific, B40922, lot 2465062) for 1 h at RT.
• Wash sections 3x 5 min with 0.01 M PBS.
• Incubate sections with Tyramide working solution containing 100X Tyramide stock solution (Alexa 488), 100X H₂O₂ solution and 1X Reaction buffer for 10 min.
• Stop the reaction with the Reaction Stop Reagent working solution.
• Wash sections 3x 5 min with 0.01M PBS.
• Mount specimens in Fluoroshield (Sigma, F6182, lot MKCB0153V).
• Acquire images with a fluorescence microscope and appropriate filter settings for AF488, e.g. Leica DM 6000B fluorescence microscope equipped with a digital camera at 40x magnification.

Independent Validation (WB)

Validierung #104497 (Western Blotting)

by: Prof. Merighi, Laboratory of Neurobiology, Department of Veterinary Sciences, University of Turin

No.: #104497

Datum: 15.03.2023

Antigen: IDE

Chargennummer: 9C07M588

Validierte Anwendung: Western Blotting

Positivkontrolle:

Adult mouse brain, cerebellum, and liver

Bewertung:

Passed. The IDE antibody (AA 491-590) ABIN723680 works in WB at 1:1000 concentrations with sensitive ECL substrate.

Validierungsbilder

Staining of IDE positive cells in the adult mouse liver with ABIN723680.

Western blot detection of IDE (110 kDa) in adult mouse brain (1), cerebellum (2), and liver (3) tissue homogenates with ABIN723680.

Protokoll

Primärantikörper: ABIN723680

Sekundärantikörper: HRP-conjugated mouse anti-rabbit

Full Protocol:

• Homogenize tissues with cold lysis buffer containing 50 mM Tris HCl, 150 mM NaCl, 1% Triton X-100, 1 mM EDTA, and 1% protease inhibitor (Sigma P8340) using an ultrasonic homogenizer (MSE, SoniPrep 150) with 16 amplitude, 20 s on, 10 s off pulse for 60 s.
• Centrifuge tissue homogentates at 13,000 rpm for 20 min at 4 °C.
• Collect supernatants and Determine total protein content using a Bradford assay.
• Denature 50 µg of total protein for 5 min at 90 °C and subsequently separate them on a denaturing 12% PAGE-SDS gel alongside a Precision Plus Protein Dual Color Standard (Bio-Rad, 160374).
• Electro-transfer proteins onto nitrocellulose membrane (Amerscham Biosciences, RPN203D) ON in the cold room.
• Wash membrane 3x for 10 mon with 0.01 M PBS containing 0.1% Tween-20 (PBST).
• Block membrane with PBST containing 2% bovine serum albumin for 1 h at RT.
• Incubate membrane with primary rabbit anti-IDE antibody (antibodies-online, ABIN723680, lot 9C07M588) diluted 1:1,000 in PBST ON at 4 °C.
• Wash membrane 3x 10 min with PBST.
• Incubate membrane with secondary HRP-conjugated mouse anti-rabbit IgG (Sigma, A1949) diluted 1:4,000 in PBST for 1 h at RT.
• Wash membrane 3x 10 min with PBST.
• Visualize proteins with WesternBright Sirius HRP substrate (Advansta, K-12043) using a ChemiDoc Imaging System.

Handhabung

Format: Liquid

Konzentration: 1 μg/μL

Buffer: 0.01M TBS( pH 7.4) with 1 % BSA, 0.02 % Proclin300 and 50 % Glycerol.

Konservierungsmittel: ProClin

Vorsichtsmaßnahmen: This product contains ProClin: a POISONOUS AND HAZARDOUS SUBSTANCE, which should be handled by trained staff only.

Lagerung: 4 °C,-20 °C

Informationen zur Lagerung: Shipped at 4°C. Store at -20°C for one year. Avoid repeated freeze/thaw cycles.

Haltbarkeit: 12 months

Details zu IDE

Target: IDE (Insulin-Degrading Enzyme (IDE))

Andere Bezeichnung: IDE (IDE Produkte)

Synonyme: zgc:162603 antikoerper, IDE antikoerper, 1300012G03Rik antikoerper, 4833415K22Rik antikoerper, AA675336 antikoerper, AI507533 antikoerper, INSDEGM antikoerper, INSULYSIN antikoerper, insulin degrading enzyme antikoerper, insulin-degrading enzyme antikoerper, metallopeptidase antikoerper, IDE antikoerper, ide antikoerper, LOC591315 antikoerper, CC1G_01115 antikoerper, Bm1_26615 antikoerper, PTRG_10875 antikoerper, SJAG_01430 antikoerper, EBI_21700 antikoerper, VDBG_06702 antikoerper, LOAG_08082 antikoerper, PGTG_04982 antikoerper, Tsp_00424 antikoerper, Ide antikoerper

Hintergrund:

Synonyms: INSULYSIN, Insulin-degrading enzyme, Abeta-degrading protease, Insulin protease, Insulinase, IDE

Background: Plays a role in the cellular breakdown of insulin, IAPP, glucagon, bradykinin, kallidin and other peptides, and thereby plays a role in intercellular peptide signaling. Degrades amyloid formed by APP and IAPP. May play a role in the degradation and clearance of naturally secreted amyloid beta-protein by neurons and microglia.

Gen-ID: 3416

UniProt: P14735

Pathways: SARS-CoV-2 Protein Interaktom