CITED2 anticorps

N° du produit ABIN6291914

Détail du produit anti-CITED2 anticorps

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

Reactivité: Humain, Souris, Rat

Hôte: Lapin

Clonalité: Polyclonal

Conjugué: Cet anticorp CITED2 est non-conjugé

Application: Western Blotting (WB)

Purification: Affinity purification

Immunogène: Recombinant protein of human CITED2

Isotype: IgG

Information d'application

Indications d'application: WB 1:500 - 1:2000

Restrictions: For Research Use only

Independent Validation (WB)

Validation #104257 (Western Blotting)

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

No.: #104257

Date: 10.02.2020

Antigène: CITED2

Numéro du lot: 469797550101

Application validée: Western Blotting

Contrôle positif:

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

Contrôle négative:

Transient transfection with empty vector

Conclusion:

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

Validation Images

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.

Protocole

Anticorps primaire: ABIN6291914

Anticorps secondaire: 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).

Notes:

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.

Stockage

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

Agent conservateur: Sodium azide

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

Stock: -20 °C

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

Détails sur CITED2

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

Autre désignation: CITED2 (CITED2 Produits)

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

Sujet: 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.

Poids moléculaire: 28.497 kDa

ID gène: 10370

UniProt: Q99967

Pathways: Tube Formation