Product Name:
C-EBPa-pT226+pT230
Product Number:
ab-pn575
Target Full Name: CCAAT/enhancer-binding protein alpha
Product Type Specific: C-EBP-alpha phosphosite-specific antibody
Antibody Code: PN575
Antibody Target Type: Phosphosite-specific
Antibody Phosphosite: T226+T230
Protein UniProt: P49715
Protein SigNET: C-EBP
Antibody Type: Polyclonal
Antibody Host Species: Rabbit
Antibody Immunogen Source: Synthetic phosphopeptide patterned after human C-EBPa
Antibody Immunogen Sequence: CGHP(pT)PPP(pT)PVP
Antibody Immunogen Description: Corresponds to amino acid residues C223 to P233. The effects of T226 and T230 phosphorylations are unclear. Their phosphorylation has been reported to both inhibit and activate transcriptional activity. Neither of these two phosphorylation sites have apparently been reported from high throughput mass spectrometry, although there are several literature reports for their phosphorylation. C-EBP-alpha is known to be phosphorylated in vitro at T226 by BARK2 (GRK3, ADRBK2), GSK3a, and GSK3b, and at T230 by GSK3a.
Production Method: The immunizing peptide was produced by solid phase synthesis on a multipep peptide synthesizer and purified by reverse-phase hplc chromatography. Purity was assessed by analytical hplc and the amino acid sequence confirmed by mass spectrometry analysis. This peptide was coupled to KLH prior to immunization into rabbits. New Zealand White rabbits were subcutaneously injected with KLH-coupled immunizing peptide every 4 weeks for 4 months. The sera from these animals was applied onto an agarose column to which the immunogen peptide was thio-linked. Antibody was eluted from the column with 0.1 M glycine, pH 2.5. Subsequently, the antibody solution was neutralized to pH 7.0 with saturated Tris.
Antibody Modification: Unconjugated. Contact KInexus if you are interest in having the antibody biotinylated or coupled with fluorescent dyes.
Antibody Concentration: 1 mg/ml
Storage Buffer: Phosphate buffered saline pH 7.4, 0.05% Thimerasol
Storage Conditions: For long term storage, keep frozen at -40°C or lower. Stock solution can be kept at +4°C for more than 3 months. Avoid repeated freeze-thaw cycles.
Product Use: Western blotting | Antibody microarray
Antibody Dilution Recommended: 2 µg/ml for immunoblotting
Antibody Species Reactivity: This antibody detects the target phosphoprotein in the following species due to conservation of amino acid sequence: Human | Chimpanzee | Rhesus macaque | Rat | Mouse | Frog.
Scientific Background: C/EBP-alpha (CCAAT/enhancer-binding protein alpha) is a bZIP transcription factor that coordinates proliferation arrest and the differentiation of myeloid progenitors, adipocytes, hepatocytes, and cells of the lung and the placenta. C/EBP-alpha and -beta play essential and redundant functions during early embryogenesis. It forms homodimers or heterodimers with C/EBP-beta and -gamma. It is required for postnatal maintenance of systemic energy homeostasis and lipid storage. It downregulates the expression of genes that maintain cells in an undifferentiated and proliferative state through E2F1 repression, which is critical for its ability to induce adipocyte and granulocyte terminal differentiation. Proliferation arrest also depends on a functional binding to SWI/SNF complex. In liver, C/EBP-alpha regulates gluconeogenesis and lipogenesis through different mechanisms. To regulate gluconeogenesis, it functionally cooperates with FOXO1 binding to IRE-controlled promoters and regulates the expression of target genes such as PCK1 or G6PC. To modulate lipogenesis, it interacts and transcriptionally synergizes with SREBF1 in promoter activation of specific lipogenic target genes such as ACSS2. In adipose tissue, it seems to act as FOXO1 coactivator accessing to adiponectin promoter through FOXO1 binding sites. It is mutated in 9% of patients with acute myeloid leukemia (AML). Human isoform 3 (p30) promotes the maintenance of an undifferentiated cellular state. Mutations that favor expression of isoform 3 are the most common type of C/EBPa mutation in AML. Isoform 3 preferentially interacts with Wdr5, inhibiting differentiation and promoting the development of SET/MLL (SET-domain/mixed-lineage leukemia).Human isoform 4 directly and specifically enhances ribosomal DNA transcription interacting with RNA polymerase I-specific cofactors and inducing histone acetylation. This description may include information annotated by UniProt and/or Google AI.