UCP1 anticorps (AA 179-296)

N° du produit ABIN2905048

Détail du produit anti-UCP1 anticorps

Antigène: UCP1 (Uncoupling Protein 1 (Mitochondrial, Proton Carrier) (UCP1))

Épitope: AA 179-296

Reactivité: Souris

Hôte: Lapin

Clonalité: Polyclonal

Conjugué: Cet anticorp UCP1 est non-conjugé

Application: Western Blotting (WB), Immunohistochemistry (IHC), Immunoprecipitation (IP), Immunofluorescence (fixed cells) (IF/ICC)

Purification: Antigen-specific affinity chromatography followed by Protein A affinity chromatography

Immunogène: UCP1 (Pro179-Leu296)

Information d'application

Indications d'application: Optimal working dilution should be determined by the investigator.

Restrictions: For Research Use only

Independent Validation (WB)

Validation #103357 (Western Blotting)

by: Mivechi lab, Georgia Cancer Center, Augusta University

No.: #103357

Date: 03.10.2018

Antigène: UCP1

Numéro du lot: AB1808U769

Application validée: Western Blotting

Contrôle positif:

Cold exposed mouse brown adipose tissue (BAT)

Contrôle négative:

Mouse liver tissue

Conclusion:

Passed. ABIN3056733 can detect mouse UCP1 protein in immunoblotting analysis.

Validation Images

Lysates from mitochondrial fraction isolated from mouse liver tissue (negative control) or brown adipose tissue (BAT) without (22°C) or with (4°C) cold exposure for 6h were resolved by SDS-PAGE gel electrophoresis followed by Western blotting analysis using UCP1 primary antibody ABIN3056733.

Protocole

Anticorps primaire: ABIN3056733

Anticorps secondaire: donkey anti-rabbit HRP-conjugated antibody (Jackson Immuno Research, 711-035-152)

Full Protocol:

• Homogenize mouse brown fat tissue in 2ml mitochondria isolation buffer (70mM sucrose, 210mM mannitol, 5mM HEPES, 1mM EGTA, pH7.2, and 0.5% BSA).
• Centrifuge the homogenates for 10min at 800g at 4°C.
• Centrifuge the supernatant again at 8000g for 10min at 4°C.
• Lyse the resulting mitochondria fraction in RIPA buffer containing protease inhibitor cocktail (Pierce Protease Inhibitor, A32965, lot TA2492391).
• Determined protein concentration with Pierce BCA Protein Assay Kit (Pierce, 23225, lot OA180049).
• Separate 50µg of total protein per lane on a denaturing 12% SDS-PAGE gel (Bio-Rad, 1610156, lot 64144809) in a Mini Protean Tetra Cell for 2h at 80V.
• Transfer proteins onto Immobilon-P PVDF Membrane (Millipore, IPVH00010) using Mini Trans Blot Cell system (Bio-Rad, 1703930).
• Block membrane with 5% milk in PBS-T for 1h at RT.
• Incubate membrane with primary rabbit anti-UCP1 antibody (antibodies-online, ABIN3056733, lot AB1808U769) diluted to 1µg/ml ON at 4°C.
• Wash membrane 3x for 10min with PBS-T.
• Incubate membrane with secondary donkey anti-rabbit HRP-conjugated antibody (Jackson Immuno Research, 711-035-152) for 1h at RT.
• Wash membrane 3x for 10min with PBS-T.
• Incubate membrane Pierce ECL Western Blotting Substrate working solution (Thermo Scientific, 1859701, lot TB263310) for 1min at RT. Expose membrane to x-ray film for different amount of time.

Notes:

The UCP1 antibody ABIN3056733 reveals a specific protein band in positive control samples (cold exposed mouse brown fat tissue), but not in negative control samples (liver tissue).

Stockage

Format: Liquid

Concentration: 500 μg/mL

Buffer: 0.01M PBS,  pH 7.4, containing 0.05 % Proclin-300, 50 % glycerol.

Agent conservateur: ProClin

Précaution d'utilisation: This product contains ProClin: a POISONOUS AND HAZARDOUS SUBSTANCE which should be handled by trained staff only.

Stock: 4 °C,-20 °C

Stockage commentaire: Store at 4 °C for frequent use. Aliquot and store at -20 °C for one year. Avoid repeated freeze/thaw cycles.

Détails sur UCP1

Antigène: UCP1 (Uncoupling Protein 1 (Mitochondrial, Proton Carrier) (UCP1))

Autre désignation: Uncoupling Protein 1, Mitochondrial (UCP1 Produits)

Synonymes: anticorps ARABIDOPSIS THALIANA PLANT UNCOUPLING MITOCHONDRIAL PROTEIN 1, anticorps ARABIDOPSIS THALIANA UNCOUPLING PROTEIN 1, anticorps ATPUMP1, anticorps ATUCP1, anticorps UCP, anticorps UCP1, anticorps UNCOUPLING PROTEIN 1, anticorps plant uncoupling mitochondrial protein 1, anticorps ucp, anticorps slc25a7, anticorps SLC25A7, anticorps AI385626, anticorps Slc25a7, anticorps Ucp, anticorps Ucpa, anticorps Uncp, anticorps plant uncoupling mitochondrial protein 1, anticorps uncoupling protein 1, anticorps uncoupling protein 1 L homeolog, anticorps uncoupling protein 1 (mitochondrial, proton carrier), anticorps PUMP1, anticorps UCP1, anticorps ucp1.L, anticorps ucp1, anticorps Ucp1

UniProt: P12242

Pathways: Proton Transport, Brown Fat Cell Differentiation