Notch 2 (NOTCH2) (Internal Domain) antibody

Antigen: Notch 2 (NOTCH2)

Synonyms: hN2, AGS2, N2, AI853703, NOTCH2

Epitope: Internal Domain

Clonality: Polyclonal

Host: Rabbit

Reactivity: Mouse (Murine), Human, Rat (Rattus)

Application: ELISA, Western Blotting (WB), Immunoprecipitation (IP)

Catalog no.: ABIN104900

Additional Information

Alternative name: NOTCH 2 (Internal Domain)

Gene ID: NP_077719

Immunogen: This whole rabbit serum was prepared by repeated immunizations with a synthetic peptide corresponding toamino acid residues 2396-2409 of human Notch 2 (the total protein i Protein Sequence: Human Notch 2 (preprotein), 2471 aa, predicted MW 110.0 kDa 1 mpalrpallw allalwlcca apahalqcrd gyepcvnegm cvtyhngtgy ckcpegflge 61 ycqhrdpcek nrcqnggtcv aqamlgkatc rcasgftged cqystshpcf vsrpclnggt 121 chmlsrdtye ctcqvgftgk ecqwtdacls hpcangstct tvanqfsckc ltgftgqkce 181 tdvnecdipg hcqhggtcln lpgsyqcqcp qgftgqycds lyvpcapspc vnggtcrqtg 241 dftfecnclp gfegstcern iddcpnhrcq nggvcvdgvn tyncrcppqw tgqfctedvd 301 ecllqpnacq nggtcanrng gygcvcvngw sgddcsenid dcafasctpg stcidrvasf … 2101 kksrrpsaks tmptslpnla keakdakgsr rkkslsekvq lsessvtlsp vdslesphty 2161 vsdttsspmi tspgilqasp npmlataapp apvhaqhals fsnlhemqpl ahgastvlps 2221 vsqllshhhi vspgsgsags lsrlhpvpvp adwmnrmevn etqynemfgm vlapaegthp 2281 giapqsrppe gkhittprep lppivtfqli pkgsiaqpag apqpqstcpp avagplptmy 2341 qipemarlps vafptammpq qdgqvaqtil payhpfpasv gkyptppsqh syassnaaer 2401 tpshsghlqg ehpyltpspe spdqwssssp hsasdwsdvt tsptpggagg gqrgpgthms 2461 epphnnmqvy as 2471 aa). A residue of cysteine was added to the aminoterminal end to facilitate coupling.

Format: Liquid (sterile filtered)

Description: Notch is synthesized in the endoplasmic reticulum as an inactive form which is proteolyticallycleaved by a furin-like convertase (S1 cleavage) in the trans-golgi network before it reaches the plasma membrane to yield anactive, ligand-accessible form. Cleavage results in a C-terminal fragment N(TM) and a N-terminal fragment N(EC). Followingligand binding, it is cleaved (S2 cleavage) by TNF-alpha converting enzyme (TACE) to yield a membrane-associatedintermediate fragment called Notch extracellular truncation (NEXT). This fragment is then cleaved by presenilin-dependentgamma-secretase (S3 cleavage) to release the intracellular domain (NICD) from the membrane. Notch functions as a receptorfor membrane-bound ligands Jagged1, Jagged2 and Delta1 to regulate cell-fate determination. Upon ligand activation throughthe released notch intracellular domain (NICD) it forms a transcriptional activator complex with RBP-J kappa and activatesgenes of the enhancer of split locus. Affects the implementation of differentiation, proliferation and apoptotic programs.

Specificity: This antiserum is directed against human NOTCH 2. The peptide sequence shows 100% alignmentwith human, dog and chimpanzee sequence. Only one (1) amino acid difference is found in mouse and this change is non-conservative. Based on the sequence we expect this antibody to react as well with rat and mouse NOTCH 2. No specificinformation is available for other reactivities.

Application Details

Application Notes: This antibody has been tested for use in ELISA and western blot. Specific conditions for reactivityshould be optimized by the end user. For western blot experiments, one can expect a band of approx 110 kDa in size corresponding to active Notch 2 in the appropriate cell lysate.

Concentration: 85 mg/ml (by Refractometry)

Buffer: Stabilizer: None. Preservative: 0.1% (w/v) Sodium Azide. Buffer: 0.02 M Potassium Phosphate, 0.15 M Sodium Chloride, pH 7.2.

Storage: Store vial at -20° C prior to opening. Dilute only prior to immediate use. For extended storage aliquotcontents and freeze at -20° C or below. Avoid cycles of freezing and thawing. Expiration date is one (1) year from date ofopening if stored frozen.

Research Area: Receptors, Signaling, Inflammation, Immunology

Restrictions: For Research Use only

Images

Western blot using antibodies-online's Anti-Notch 2 (intra) antibody shows detection of a band at ~110 kDa corresponding to active Notch 2 protein (arrowhead). Western Blot analysis was performed for Notch 2 expression using 100ug of total protein lysate obtained from human mesothelial SV40 cells transfected with a plasmid encoding a constitutively active Notch 2 (intra cellular Notch 2). Lanes 1-3 contain lysate 24 h (1), 48 h (2), and 72 h (3) post transfection. Lanes 4-6 are the corresponding control cells (untransfected) taken at similar time points. The band at about 110kD represents active Notch 2. This band is not seen in the control cell. The intracellular domain of Notch 2 has a predicted band size of 110kD, corresponding to this band. Protein cell lysates were run on a 10% SDS-page gel, blotted onto Hybond C membrane, blocked overnight in PBS-Tween 20 supplemented with 5% Non-fat Milk and probed with anti-Notch 2 at a 1:400 dilution. ECL was used as visualization method.