CITED2 antibody
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- Target See all CITED2 Antibodies
- CITED2 (Cbp/p300-Interacting Transactivator, with Glu/Asp-Rich Carboxy-terminal Domain, 2 (CITED2))
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Reactivity
- Human, Mouse, Rat
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Host
- Rabbit
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Clonality
- Polyclonal
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Conjugate
- This CITED2 antibody is un-conjugated
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Application
- Western Blotting (WB)
- Purification
- Affinity purification
- Immunogen
- Recombinant protein of human CITED2
- Isotype
- IgG
- Top Product
- Discover our top product CITED2 Primary Antibody
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- Application Notes
- WB 1:500 - 1:2000
- Restrictions
- For Research Use only
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- by
- Dr. Pengpeng Bi Lab, Department of Genetics, Franklin College of Arts and Sciences, University of Georgia
- No.
- #104257
- Date
- 02/10/2020
- Antigen
- CITED2
- Lot Number
- 469797550101
- 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. ABIN6291914 specifically recognizes Cited2 in human HEK293 cell lysates.
- Primary Antibody
- ABIN6291914
- 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 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).
- Experimental 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.
Validation #104257 (Western Blotting)Validation ImagesFull Methods -
- Buffer
- PBS with 0.02 % sodium azide, 50 % glycerol, pH 7.3.
- Preservative
- Sodium azide
- Precaution of Use
- This product contains Sodium azide: a POISONOUS AND HAZARDOUS SUBSTANCE which should be handled by trained staff only.
- Storage
- -20 °C
- Storage Comment
- Store at -20C. Avoid freeze / thaw cycles.
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- Target
- CITED2 (Cbp/p300-Interacting Transactivator, with Glu/Asp-Rich Carboxy-terminal Domain, 2 (CITED2))
- Alternative Name
- CITED2 (CITED2 Products)
- Synonyms
- mrg1 antibody, p35srj antibody, zgc:103418 antibody, CITED2 antibody, cite2 antibody, ASD8 antibody, MRG-1 antibody, MRG1 antibody, P35SRJ antibody, VSD2 antibody, AI835299 antibody, ER154-like antibody, Mrg1 antibody, Msg2 antibody, Cbp/p300-interacting transactivator, with Glu/Asp rich carboxy-terminal domain, 2 antibody, Cbp/p300-interacting transactivator, with Glu/Asp-rich carboxy-terminal domain, 2 antibody, Cbp/p300 interacting transactivator with Glu/Asp rich carboxy-terminal domain 2 antibody, Cbp/p300-interacting transactivator 2 antibody, Cbp/p300-interacting transactivator, with Glu/Asp rich carboxy-terminal domain, 2 L homeolog antibody, cited2 antibody, CITED2 antibody, cite2 antibody, Cited2 antibody, cited2.L antibody
- Background
- 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.
- Molecular Weight
- 28.497 kDa
- Gene ID
- 10370
- UniProt
- Q99967
- Pathways
- Tube Formation
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