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C-MYB PT486 Antibody

Rabbit Polyclonal
NCI Collaboration
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Western Blot of Rabbit c-Myb pT486 antibody.  
Lane 1:  Cos7 cells were either null for c-Myb.  
Lane 2:  Cos7 cell lysates transfected with c-myb, no treatment.  
Lane 3:  Cos7 cell lysates transfected with c-Myb andtreated with okadaic acid to incude phosphorylation of c-Myb.  
Load:  35 µg per lane.
Primary antibody:  The left blot was probed with anti-c-Myb antibody, the right blot was probed with anti-c-Myb pT486 antibody at 1:2500 for overnight at 4°C.
Secondary antibody:  IRDye800™ rabbit secondary antibody at 1:10,000 for 45 min at RT.
Block:  PBS-Tween (0.05%) overnight at 4°C.
Predicted/Observed size:  75, 98, and 125kDa corresponding to overexpressed c-Myb (arrow): wild-type, with one attached SUMO molecule, and with two attached SUMO molecules, respectively.
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Number:
600-401-C47
Size:
100 µg
Price:
$309.00
Shipping:
$20.00
Handling:
$20.00
Availability:
Ships next business day.

C-MYB PT486 Antibody Properties

Description
Anti-c-Myb pT486 (RABBIT) Antibody - 600-401-C47
Host
Rabbit
Target Species
Mouse
Known Cross Reactivity
Mouse
Clonality
Polyclonal
Application
ELISA : 1:5,000-1:16,000
Western Blot : 1:2,500
Other Dilution: User Optimized
Physical State
Liquid (sterile filtered)
Shipping Condition
Dry Ice
Format
IgG
Label
Concentration
1.51 by UV absorbance at 280 nm
Buffer
0.02 M Potassium Phosphate, 0.15 M Sodium Chloride, pH 7.2
Preservative
0.01% (w/v) Sodium Azide
Stabilizer
None
GeneName
MYB
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C-MYB PT486 Antibody Description

Background
This antibody is designed, produced, and validated as part of a collaboration between Rockland and the National Cancer Institute (NCI) and is suitable for Cancer, Immunology and Nuclear Signaling research. The transcription factor c-Myb plays an important role in the development of all hematopoietic lineages during definitive hematopoiesis. Its critical role in hematopoiesis has been first demonstrated in experiments with targeted disruption of the c-Myb gene where deletion of both alleles of the c-Myb gene was embryonally lethal due to severe anemia. c-Myb is a short-lived sequence specific DNA-binding protein that regulates transcription of several important genes involved directly in cellular processes such as proliferation, differentiation, and apoptosis. Post-translational modifications such as phosphorylation, acetylation, ubiquitination, and conjugation of SUMO proteins are crucial for modulation of the transactivation activity of c-Myb. Several phosphorylation sites serve to regulate c-Myb activity.
Synonyms
Avian myeloblastosis viral (v-myb) oncogene homolog antibody, C myb antibody, MYB antibody, v-myb avian myeloblastosis viral oncogene homolog antibody
Immunogen
This affinity purified antibody was prepared from whole rabbit serum produced by repeated immunizations with a synthetic peptide corresponding to an internal region of mouse c-Myb protein surrounding amino acid residue 486.
Immunogen Type
Peptide
Post Translational Modification
Phosphorylation
Storage Condition
Store vial at -20° C prior to opening. Aliquot contents and freeze at -20° C or below for extended storage. Avoid cycles of freezing and thawing. Centrifuge product if not completely clear after standing at room temperature. This product is stable for several weeks at 4° C as an undiluted liquid. Dilute only prior to immediate use.
Application Note
This affinity purified antibody has been tested for use in ELISA and western blotting using M1 cells which express endogenous wild-type c-Myb protein and in COS7 cells transfected with c-Myb.  Specific conditions for reactivity and detection of phosphorylated c-Myb should be optimized by the end user.  Expect a band approximately ~75 kDa in size corresponding to phosphorylated c-Myb by western blotting in the appropriate cell lysate or extract.
Purity/Specifity
This affinity-purified antibody is directed against the phosphorylated form of mouse c-Myb protein at the pT486 residue. Antiserum was first purified against the phosphorylated form of the immunizing peptide.  The resultant affinity purified antibody was then cross adsorbed against the non-phosphorylated form of the immunizing peptide.  Reactivity occurs against mouse c-Myb pT486 protein and the antibody is specific for the phosphorylated form of the protein.  This antibody reacts with endogenous c-Myb protein and also SUMOylated overexpressed c-Myb protein in transfected Cos7 cells. Reactivity with non-phosphorylated mouse c-Myb is minimal by ELISA and western blot. A BLAST analysis was used to suggest reactivity with c-Myb from mouse based on a 100% homology with the immunizing sequence.  Expect reactivity with c-Myb from rat, pig, human, horse, platypus, opossum, orangutan, spider monkey, bonobo, gorilla, and chimpanzee; with a v-myb myeloblastosis viral oncogene homolog, all isoforms (also avian version) from human; and with a protooncogene c-myb, and a v-myb myeloblastosis viral oncogene homolog (avian), both from cattle, all based on a 93% homology with the immunizing sequence.  Cross-reactivity with c-Myb from other sources has not been determined.
Disclaimer Note-General
This product is for research use only and is not intended for therapeutic or diagnostic applications. Please contact a technical service representative for more information. All products of animal origin manufactured by Rockland Immunochemicals are derived from starting materials of North American origin. Collection was performed in United States Department of Agriculture (USDA) inspected facilities and all materials have been inspected and certified to be free of disease and suitable for exportation. All properties listed are typical characteristics and are not specifications. All suggestions and data are offered in good faith but without guarantee as conditions and methods of use of our products are beyond our control. All claims must be made within 30 days following the date of delivery. The prospective user must determine the suitability of our materials before adopting them on a commercial scale. Suggested uses of our products are not recommendations to use our products in violation of any patent or as a license under any patent of Rockland Immunochemicals, Inc. If you require a commercial license to use this material and do not have one, then return this material, unopened to: Rockland Inc., P.O. BOX 326, Gilbertsville, Pennsylvania, USA.
General Reference
M.Sramko, J.Markus, J.Kabat, L.Wolff, and J.Bies. (2006) Stress-induced Inactivation of the c-Myb Transcription Factor through Conjugation of SUMO-2/3 Proteins. J Biol Chem 281: 40065. J. Bies, J.Markus, and L.Wolff, (2002) Covalent Attachment of the SUMO-1 Protein to the Negative Regulatory Domain of the c-Myb Transcription Factor Modifies Its Stability and Transactivation Capacity. J Biol Chem 277: 8999.
Specific Reference
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Product Type
GFP and Epitopes;
Applications
ELISA, WB, IHC, IP
Reacts With
C-terminal, N-terminal, and internal tagged GST fusion proteins
Catalog Number
200-301-200

Product Type
Primary Antibodies;
Applications
ELISA, WB, IHC, FC
Reacts With
human, mouse, rat, chimpanzee
Catalog Number
200-301-401

Product Type
Primary Antibodies;
Applications
ELISA, WB
Reacts With
Mouse
Catalog Number
600-401-C47S

Product Type
Secondary Antibodies;
Applications
ELISA, WB, IHC
Catalog Number
611-1302

Product Label

 

Description
Unconjugated
Color
Conjugation Reference
Molecular Weight
Excitation Wavelength
Conjugation Chemistry