CITED2 Antikörper

Produktnummer ABIN6291914

Produktdetails anti-CITED2 Antikörper

Target: CITED2 (Cbp/p300-Interacting Transactivator, with Glu/Asp-Rich Carboxy-terminal Domain, 2 (CITED2))

Reaktivität: Human, Maus, Ratte

Wirt: Kaninchen

Klonalität: Polyklonal

Konjugat: Dieser CITED2 Antikörper ist unkonjugiert

Applikation: Western Blotting (WB)

Aufreinigung: Affinity purification

Immunogen: Recombinant protein of human CITED2

Isotyp: IgG

Anwendungsinformationen

Applikationshinweise: WB 1:500 - 1:2000

Beschränkungen: Nur für Forschungszwecke einsetzbar

Independent Validation (WB)

Validierung #104257 (Western Blotting)

by: Dr. Pengpeng Bi Lab, Department of Genetics, Franklin College of Arts and Sciences, University of Georgia

No.: #104257

Datum: 10.02.2020

Antigen: CITED2

Chargennummer: 469797550101

Validierte Anwendung: Western Blotting

Positivkontrolle:

HEK293 cells overexpressing murine Cited2 (NCBI protein ID: NP_034958, 269 amino acids) from a transiently transfected expression plasmid

Negativkontrolle:

Transient transfection with empty vector

Bewertung:

Passed. ABIN6291914 specifically recognizes Cited2 in human HEK293 cell lysates.

Validierungsbilder

Western blot analysis of cell lysates from HEK293 cells transfected with an empty vector (EV) or a murine Cited2 expression plasmid (mCited2). Staining was performed using ABIN6291914 (top), or an alpha-tubulin loading control (bottom) antibody.

Protokoll

Primärantikörper: ABIN6291914

Sekundärantikörper: Goat anti-Rabbit IgG (H+L), Superclonal Recombinant Secondary Antibody, HRP (Invitrogen, A27036, lot 20310171)

Full Protocol:

• Grow human HEK293 cells (Cell Biolabs, RV-101) in DMEM medium(Sigma, D5796) supplemented with FBS (Gemini, A59G00J) and Pen Strep (Gibco, 15140-122, lot 1999384), at 37°C and 5% CO₂ in 2 mL medium on a 6 well plate to 80% confluency.
• Transfect cells with 2ug per well pMXs-Puro murine Cited2 expression plasmid using PEI (Polysciences, 23966-1) following the manufacturer´s instructions.
• Lyse 1x10⁶ cells in 100µl per well cold RIPA (Sigma, R0278).
• Determine total protein content of the lysates using Pierce BCA Protein Assay Kit(Thermo Fisher, 23225).
• Mix 40µg of total protein with in 10µL 4x Laemmli Sample Buffer (Biorad, 1610747) and subsequently separate them on a denaturing 12% PAGE gel (Biorad, 1610156) in a Biorad electrophoresis chamber for 1h at 100V.
• Transfer proteins onto Immobilon-PSQ PVDF Membrane (Sigma, ISEQ00010, lot R8CA8256B) with a Western blotting system for 1h at 110V.
• Block the membrane with blocking buffer (5% non-fat milk in TBST) for 1h at RT.
• Incubation with primary rabbit anti-CITED2 antibody (antibodies-online, ABIN6291914, lot 469797550101) diluted 1:300 in blocking buffer ON at 4°C.
• Wash membrane 3x for 15min with TBST.
• Incubation with Goat anti-Rabbit IgG (H+L), Superclonal Recombinant Secondary Antibody, HRP (Invitrogen, A27036, lot 20310171) diluted 1:2000 in blocking buffer for 1h at RT.
• Wash membrane 5x for 15min with TBST.
• Reveal protein bands using SuperSignal West Dura (Thermo Scientific, 34075, TG268239) on a UVP ChemStudio touch (Analytik Jena, 84997092802).

Anmerkungen:

The Cited2 antibody ABIN6291914 reveals a protein of the expected molecular weight of antigen in lysates of human HEK293 cells expressing murine Cited2. The protein band is only visible in the positive but not the negative control.

Handhabung

Buffer: PBS with 0.02 % sodium azide, 50 % glycerol,  pH 7.3.

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: Store at -20C. Avoid freeze / thaw cycles.

Details zu CITED2

Target: CITED2 (Cbp/p300-Interacting Transactivator, with Glu/Asp-Rich Carboxy-terminal Domain, 2 (CITED2))

Andere Bezeichnung: CITED2 (CITED2 Produkte)

Synonyme: mrg1 antikoerper, p35srj antikoerper, zgc:103418 antikoerper, CITED2 antikoerper, cite2 antikoerper, ASD8 antikoerper, MRG-1 antikoerper, MRG1 antikoerper, P35SRJ antikoerper, VSD2 antikoerper, AI835299 antikoerper, ER154-like antikoerper, Mrg1 antikoerper, Msg2 antikoerper, Cbp/p300-interacting transactivator, with Glu/Asp rich carboxy-terminal domain, 2 antikoerper, Cbp/p300-interacting transactivator, with Glu/Asp-rich carboxy-terminal domain, 2 antikoerper, Cbp/p300 interacting transactivator with Glu/Asp rich carboxy-terminal domain 2 antikoerper, Cbp/p300-interacting transactivator 2 antikoerper, Cbp/p300-interacting transactivator, with Glu/Asp rich carboxy-terminal domain, 2 L homeolog antikoerper, cited2 antikoerper, CITED2 antikoerper, cite2 antikoerper, Cited2 antikoerper, cited2.L antikoerper

Hintergrund: The protein encoded by this gene inhibits transactivation of HIF1A-induced genes by competing with binding of hypoxia-inducible factor 1-alpha to p300-CH1. Mutations in this gene are a cause of cardiac septal defects. Alternatively spliced transcript variants encoding multiple isoforms have been observed for this gene.

Molekulargewicht: 28.497 kDa

Gen-ID: 10370

UniProt: Q99967

Pathways: Tube Formation