CITED2 antibody (N-Term)

Catalog No. ABIN2776451

This independently validated Rabbit Polyclonal antibody specifically detects CITED2 in WB and IHC. It exhibits reactivity toward Human, Mouse, Rat, Cow, Guinea Pig, Dog and Sheep.

Quick Overview for CITED2 antibody (N-Term) (ABIN2776451)

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

Reactivity: Human, Mouse, Rat, Cow, Guinea Pig, Dog, Sheep

Host: Rabbit

Clonality: Polyclonal

Conjugate: This CITED2 antibody is un-conjugated

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

Product Details anti-CITED2 Antibody

Binding Specificity: N-Term

Sequence: HIHYGAGNMN ATSGIRHAMG PGTVNGGHPP SALAPAARFN NSQFMGPPVA

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

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

Purification: Affinity Purified

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

Application Details

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

Comment:

Antigen size: 270 AA

Restrictions: For Research Use only

Independent Validation (WB)

Validation #104256 (Western Blotting)

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

No.: #104256

Date: 02/10/2020

Antigen: CITED2

Lot Number: QC11500

Method validated: Western Blotting

Positive Control:

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

Negative Control:

Transient transfection with empty vector

Notes:

Passed. ABIN2776451 specifically recognizes Cited2 in human HEK293 cell lysates.

Validation Images

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

Full Methods

Primary Antibody: ABIN2776451

Secondary Antibody: 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).

Experimental Notes:

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.

Handling

Format: Liquid

Concentration: Lot specific

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

Preservative: Sodium azide

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

Handling Advice: Avoid repeated freeze-thaw cycles.

Storage: -20 °C

Storage Comment: 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.

Target Details for CITED2

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

Alternative Name: CITED2

Background: 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

Molecular Weight: 28 kDa

Gene ID: 10370

NCBI Accession: NM_006079, NP_006070

UniProt: Q99967

Pathways: Tube Formation