AQP3 Antikörper (C-Term)

Produktnummer ABIN863208

Dieses Anti-AQP3-Antikörper ist ein Kaninchen Polyklonal-Antikörper zur Detektion von AQP3 in WB, IHC, IF und ICC. Geeignet für Ratte. Unabhängig validiert für die Verwendung in Immunofluorescence.

Kurzübersicht für AQP3 Antikörper (C-Term) (ABIN863208)

Target: AQP3 (Aquaporin 3 (Gill Blood Group) (AQP3))

Reaktivität: Ratte

Wirt: Kaninchen

Klonalität: Polyklonal

Konjugat: Dieser AQP3 Antikörper ist unkonjugiert

Applikation: Western Blotting (WB), Immunohistochemistry (IHC), Immunofluorescence (IF), Immunocytochemistry (ICC)

Produktdetails anti-AQP3 Antikörper

Bindungsspezifität: C-Term

Spezifität: Detects ~31.5 kDa. May detect larger glycosylated bands ~35-50 kDa.

Kreuzreaktivität: Human, Maus, Ratte

Aufreinigung: Protein A Purified

Immunogen: Produced against the C-terminal peptide (Sequence N-CHLEQPPPSTEAENVKLAHMKHKEQI) of rat aquaporin 3

Anwendungsinformationen

Applikationshinweise:

• WB (1:2000)
• IHC (1:200)
• ICC/IF (1:400)
• optimal dilutions for assays should be determined by the user.

Kommentare:

0.5 μg/ml of ABIN863208 was sufficient for detection of aquaporin 3 in 10 μg of rat kidney tissue lysate by colorimetric immunoblot analysis using Goat anti-rabbit IgG:HRP as the secondary antibody.

Beschränkungen: Nur für Forschungszwecke einsetzbar

Independent Validation (IF)

Validierung #102635 (Immunofluorescence)

by: Centre de Recherche en Transplantation et Immunologie, UMR1064, Université de Nantes

No.: #102635

Datum: 14.12.2018

Antigen: AQP3

Chargennummer: 160308

Validierte Anwendung: Immunofluorescence

Positivkontrolle:

Membrane staining in compacted morula; membrane staining apical side, on early blastocysts

Negativkontrolle:

Cells not expressing AQP3 mRNA within human embryos (inner cell mass)

No primary control

Bewertung:

Passed. ABIN863208 specifically labels the targeted antigen in embryos in IF. No signal was detected in sample negative control tissue and the secondary antibody only control.

Validierungsbilder

Immunoflurescence analysis of human embryo using ABIN863208. AQP3 (right) with nuclear counterstaining (left) at indicated stages. In the top panel (morula), the staining is located at the membrane. In the bottom panel (early blastocyst), the staining is located at the membrane, in trophectoderm cells. Scale bar = 47µm.

Protokoll

Primärantikörper: ABIN863208

Sekundärantikörper: donkey anti-rabbit antibody (Life Technologies, A21206)

Full Protocol:

• Fixation
• • Fix embryos in one drop of 30µl of 4% paraformaldehyde for 5min at RT.
  • Remove embryos and wash quickly three times with PBS containing 0.1% BSA (in three drops of 30µl).
• Staining:
• • Permeabilize embryo in 300µl IF buffer for 60min at RT.
  • Incubate embryo with the primary rabbit anti-AQP3 antibody (antibodies-online, ABIN863208, 160308) diluted 1:400 in buffer ON at 4°C.
  • Rinse embryo in IF buffer quickly three times.
  • Incubate embryo with secondary donkey anti-rabbit antibody (Life Technologies, A21206) diluted 1:1000 in buffer for 2h at RT.
  • Rinse embryo in PBS-BSA.
  • At this point, the embryos can either be kept in PBS-BSA ON at 4°C or used for imaging directly on ibido µ-Slide.
  • Image acquisition on a microscope using appropriate filters, 20x oil-immersion objective.

Anmerkungen:

Labeling of human embryo with the aquaporin 3 antibody ABIN863208 shows the expected staining pattern membrane-associated outside of the blastocyst. The background staining in our conditions is a bit high and might be improved with more stringent washes or different fixation-permeabilization protocols.

Handhabung

Format: Liquid

Konzentration: 1 mg/mL

Buffer: PBS, 50 % glycerol, 0.09 % sodium azide, Storage buffer may change when conjugated

Konservierungsmittel: Sodium azide

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

Lagerung: -20 °C

Informationen zur Lagerung: -20°C

Details zu AQP3

Target: AQP3 (Aquaporin 3 (Gill Blood Group) (AQP3))

Andere Bezeichnung: Aquaporin 3

Hintergrund: Aquaporins selectively conduct water molecules in and out of the cell, while preventing the passage of ions and other solutes. Known as water channels, they are integral membrane pore proteins (1, 2). Aquaporin 3 is found in the basolateral cell membrane of principal collecting duct cells and provide a pathway for water to exit these cells (3). AQP3 gene expression is not regulated by vasopressin (4).

Gen-ID: 65133

NCBI Accession: NP_113891

UniProt: P47862