FNDC5 Antikörper (Fibronectin Type III Domain Containing 5)

Details for Product anti-FNDC5 Antibody No. ABIN1888571, Anbieter: Anmelden zum Anzeigen
Antigen
  • FRCP2
  • irisin
  • 1500001L03Rik
  • AI836596
  • C87088
  • PeP
  • Pxp
  • fndc5
  • si:dkey-264p5.1
  • zgc:158684
  • fibronectin type III domain containing 5
  • fibronectin type III domain containing 5b
  • FNDC5
  • Fndc5
  • fndc5b
Reaktivität
Human, Affe, Maus, Ratte (Rattus)
67
56
35
7
2
Wirt
Kaninchen
68
1
Klonalität
Polyklonal
Konjugat
Dieser FNDC5 Antikörper ist unkonjugiert
9
4
2
2
2
2
1
1
1
1
1
1
1
1
1
1
1
1
1
1
1
1
1
1
1
Applikation
Western Blotting (WB)
52
34
21
14
14
13
10
5
3
2
1
1
Optionen
Hersteller
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Hersteller Produkt- Nr.
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'Independent Validation' Siegel
Antigen FNDC5
Chargennummer ZA 128635
Validierte Anwendung Western Blotting
Positivkontrolle Recombinant irisin produced in E. coli, recombinant glycosylated irisin produced in HEK293 cells (described in Albrecht et al. (2015))
Negativkontrolle Blocking of specific antibody binding with recombinant protein prior to incubation
Bewertung ABIN1888571 recognizes recombinant protein but no antigen in the tested samples. This may be because of the low target protein abundance in the samples.
Primärantikörper ABIN1888571
Sekundärantikörper mouse anti-rabbit IgG TrueBlot HRP conjugated antibody (Rockland, 18-8816-33, lot 37203)
Protokoll
  • Lyse muscle tissue from human, mouse, and bovine in CellLytic lysis buffer (Sigma, C3228, lot 106M4000V). Plasma and serum samples were albumin and IgG depleted.
  • Determine total protein content of the lysates using NanoDrop ND-1000, BSA mode (Peqlab).
  • Denature 5ng of recombinant irisin, 15ng of recombinant glycosylated irisin, and lysate corresponding to 20µg of total protein for 5min at 95°C in an equal volume of 2x Laemmli SDS sample buffer and subsequently separate them on a freshly cast 12% denaturing polyacrylamide gel in a PerfectBlue mini M (Peqlab)electrophoresis chamber for 80min at 125V.
  • Transfer proteins onto Roti-PVDF 0.2 membrane (Roth, 8989.1) with a TransBlot Turbo Western blotting system (BioRad) for 10min at 25V and 1A.
  • Block the membrane with TBS with 10% RotiBlock (Roth, A151.4, lot no 027253792) for 1h at RT.
  • Incubation with primary rabbit-anti irisin antibody (antibodies-online, ABIN1888571, ZA 128635) diluted 1:3000 in TBST containing 10% RotiBlock for ON at 4°C.
  • Wash membrane 4x for 10min with TBST-buffer.
  • Incubation with secondary mouse anti-rabbit IgG TrueBlot HRP conjugated antibody (Rockland, 18-8816-33, lot 37203) diluted 1:25000 in TBST for 90min at RT.
  • Wash membrane 3x for 10min with TBST-buffer, 1x 5min with TBS, and 1x 5min with ddH2O.
  • Reveal protein bands using Super Signal West Femto (Thermo Scientific, 34095, lot no RF235326A) on a Chemocam HR-16 imager (INTAS).
Anmerkungen
  • The irisin antibody ABIN1888571 is able to detect the recombinant protein (described in Albrecht et al. (2015)) in its non-glycosylated and glycosylated form at the expected molecular weight (approximately 12.5kDa for the non-glycosylated form).

  • Antigen binding can be blocked if the ABIN1888571 was preincubated with the 3-fold amount of recombinant irisin. No comparable protein band was detected in murine, human, and bovine tissue, plasma, or serum.

  • A protein band of much larger size in muscle lysates seems to be unspecific, as it appears also after blocking. Increased unspecific binding was observed after blocking of the antibody with recombinant protein, as expected.

Validierungsbilder
Western Blotting validation image for anti-Fibronectin Type III Domain Containing 5 (FNDC5) antibody (ABIN1888571) Western blot for detection of irisin. (A) Quick Coomassie stained blot for demonstrat...
Verwendungszweck Polyclonal Antibody to detect Irisin in human, mouse, monkey and rat samples
Immunogen Recombinant Irisin
Isotyp IgG
Produktmerkmale Polyclonal Antibody to detect Irisin in human, mouse, monkey and rat samples
Reinigung Epitope affinity purified
Andere Bezeichnung Irisin (FNDC5 Antibody Abstract)
Hintergrund Irisin is a recently described exercise-induced hormone secreted by skeletal muscle in mice and humans. Irisin activates beige fat cells (beige cells have a gene expression pattern distinct from either white or brown fat and are preferentially sensitive to the polypeptide hormone Irisin). Irisin is cleaved from the type I membrane protein FNDC5 and improves systemic metabolism by increasing energy expenditure.
Synonyms: Fibronectin Type III Domain-containing Protein 5 (cleaved), Fibronectin Type III Repeat-containing Protein 2 (cleaved), FNDC5 (cleaved)
Gen-ID 252995
UniProt Q8NAU1
Pathways Hormone Activity, Brown Fat Cell Differentiation, Positive Regulation of fat Cell Differentiation
Applikations-hinweise Western Blot - 1: 1000 dilution
Beschränkungen Nur für Forschungszwecke einsetzbar
Format Liquid
Konzentration 1 mg/mL
Buffer 1 mg/mL in PBS containing 10 % glycerol and 0.02 % sodium azide.
Konservierungs-mittel Sodium azide
Vorsichtsmaßnahmen This product contains Sodium azide: a POISONOUS AND HAZARDOUS SUBSTANCE which should be handled by trained staff only.
Handhabung The antibody solution should be gently mixed before use.
Lagerung -20 °C
Haltbarkeit 12 months
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