CITED2 antibody (Cbp/p300-Interacting Transactivator, with Glu/Asp-Rich Carboxy-terminal Domain, 2) (N-Term)

Details for Product anti-CITED2 Antibody No. ABIN2776451
Target
Binding Specificity
N-Term
10
8
8
7
7
2
1
1
Reactivity
Cow, Dog, Guinea Pig, Human, Mouse, Rat, Sheep
49
27
15
4
4
2
2
2
1
1
Host
Rabbit
38
7
4
1
1
Clonality
Polyclonal
Conjugate
This CITED2 antibody is un-conjugated
4
3
3
3
3
3
1
1
1
1
1
1
Application
Immunohistochemistry (IHC), Western Blotting (WB)
37
28
14
6
3
2
1
1
1
1
Options
Immunogen The immunogen is a synthetic peptide directed towards the N terminal region of human CITED2
Blocking Peptide Blocking peptide for this product available: ABIN974999
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
Target
Alternative Name CITED2 (CITED2 Antibody Abstract)
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
Application Notes Optimal working dilutions should be determined experimentally by the investigator.
Comment

Antigen size: 270 AA

Restrictions For Research Use only
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.
Images
Image no. 1 for anti-Cbp/p300-Interacting Transactivator, with Glu/Asp-Rich Carboxy-terminal Domain, 2 (CITED2) (N-Term) antibody (ABIN2776451) WB Suggested Anti-CITED2 Antibody Titration: 0.2-1 ug/ml Positive Control: Human b...
'Independent Validation' Badge
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.

Primary Antibody ABIN2776451
Secondary Antibody Goat anti-Rabbit IgG (H+L), Superclonal Recombinant Secondary Antibody, HRP (Invitrogen, A27036, lot 20310171)
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.

Validation Images
Western Blotting validation image for anti-Cbp/p300-Interacting Transactivator, with Glu/Asp-Rich Carboxy-terminal Domain, 2 (CITED2) (N-Term) antibody (ABIN2776451) Western blot analysis using ABIN2776451 of cell lysates from HEK293 cells transfected...