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.

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

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