CITED2 Antikörper (N-Term)

Produktnummer ABIN2776451

Produktdetails anti-CITED2 Antikörper

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

Bindungsspezifität: N-Term

Reaktivität: Human, Maus, Ratte, Rind (Kuh), Hund, Meerschweinchen, Schaf

Wirt: Kaninchen

Klonalität: Polyklonal

Konjugat: Dieser CITED2 Antikörper ist unkonjugiert

Applikation: Western Blotting (WB), Immunohistochemistry (IHC)

Sequenz: HIHYGAGNMN ATSGIRHAMG PGTVNGGHPP SALAPAARFN NSQFMGPPVA

Homologie: Cow: 100%, Dog: 100%, Guinea Pig: 100%, Human: 100%, Mouse: 100%, Rat: 100%, Sheep: 100%

Produktmerkmale: This is a rabbit polyclonal antibody against CITED2. It was validated on Western Blot using a cell lysate as a positive control.

Aufreinigung: Affinity Purified

Immunogen: The immunogen is a synthetic peptide directed towards the N terminal region of human CITED2

Anwendungsinformationen

Applikationshinweise: Optimal working dilutions should be determined experimentally by the investigator.

Kommentare:

Antigen size: 270 AA

Beschränkungen: Nur für Forschungszwecke einsetzbar

Independent Validation (WB)

Validierung #104256 (Western Blotting)

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

No.: #104256

Datum: 10.02.2020

Antigen: CITED2

Chargennummer: QC11500

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. ABIN2776451 specifically recognizes Cited2 in human HEK293 cell lysates.

Validierungsbilder

Western blot analysis using ABIN2776451 of cell lysates from HEK293 cells transfected with an empty vector (EV) or a murine Cited2 expression plasmid (mCited2).

Protokoll

Primärantikörper: ABIN2776451

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 2mL medium on a 6 well plate to 80% confluency.
• Transfect cells with 2µg 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 40 µg of total protein with in 10 µL 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, ABIN2776451, lot QC11500) diluted 1:200 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 ABIN2776451 reveals a protein of the expected molecular weight of the antigen in lysates of human HEK293 cells expressing murine Cited2. The protein band is only visible in the positive but not the negative controls.

Handhabung

Format: Liquid

Konzentration: Lot specific

Buffer: Liquid. Purified antibody supplied in 1x PBS buffer with 0.09 % (w/v) sodium azide and 2 % sucrose.

Konservierungsmittel: Sodium azide

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

Handhabung: Avoid repeated freeze-thaw cycles.

Lagerung: -20 °C

Informationen zur Lagerung: For short term use, store at 2-8°C up to 1 week. For long term storage, store at -20°C in small aliquots to prevent 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: CITED2 belongs to the CITED family. It interferes with the binding of transcription factors HIF-1a and STAT2 to p300/CBP.
Alias Symbols: MRG1, P35SRJ, ASD8, VSD2
Protein Interaction Partner: UBC, ELAVL1, EP300, SMAD3, SMAD2, LHX2, HIF1A, TFAP2C, LHX3, CREBBP, TFAP2A, TBP, TFAP2B,
Protein Size: 270

Molekulargewicht: 28 kDa

Gen-ID: 10370

NCBI Accession: NM_006079, NP_006070

UniProt: Q99967

Pathways: Tube Formation