Anti-p-Ubiquitin (pSer65) Antibody, clone 2N21 ZooMAb^® Rabbit Monoclonal

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ZooMAb antibodies represent an entirely new generation of recombinant monoclonal antibodies.

Each ZooMAb antibody is manufactured using our proprietary recombinant expression system, purified to homogeneity, and precisely dispensed to produce robust and highly reproducible lot-to-lot consistency. Only top-performing clones are released for use by researchers. Each antibody is validated for high specificity and affinity across multiple applications, including its most commonly used application. ZooMAb antibodies are reliably available and ready to ship when you need them.

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Clone 2N21 is a ZooMAb rabbit recombinant monoclonal antibody that specifically detects ubiquitin phosphorylated on serine 65.

KLH-conjugated linear peptide corresponding to 13 amino acids from the ubiquitin-like 1 domain of human polyubiquitin phosphorylated on serine 65.

Anti-p-Ubiquitin (pSer65), clone 2N21 ZooMAb, Cat. No. ZRB1494, is a recombinant Rabbit monoclonal antibody to detect ubiquitin (pSer65) in Affinity Binding Assay, Flow Cytometry, Immunocytochemistry, Peptide Inhibition Assay, and Western Blotting.

Western Blotting Analysis: A 1:1,000 dilution from a representative lot detected p-Ubiquitin (pSer65) in lysate from HeLa cells transfected with PINK and treated with CCCP.

Flow Cytometry Analysis: 0.1 μg from a representative lot detected p-Ubiquitin (pSer65) in one million PC-3 cells treated with CCCP.

Peptide Inhibition Assay: Target band detection in lysate from HeLa cells transfected with PINK and treated with CCCP (15 mM, 5 h) was prevented by preblocking of a representative lot with the phosphorylated Ser 65 ubiquitin peptide, but not by corresponding non-phosphopeptide.

Immunocytochemistry Analysis: A 1:100 dilution from a representative lot detected p-Ubiquitin (pSer65) in HeLa cells transfected with PINK and treated wtih CCCP.

Affinity Binding Assay: A representative lot of this antibody bound phosphorylated serine 65 Ubiquitin peptide with a KD of 2.5 x 10-8 in an affinity binding assay.

Note: Actual optimal working dilutions must be determined by end user as specimens, and experimental conditions may vary with the end user

Polyubiquitin-B and Polyubiquitin-C (UniProt: P0CG47; P0CG48) are encoded by the UBB and UBC genes (Gene ID: 7314 and 7316) in human. The C-termini of Polyubiquitin-B (Cys229) and Polyubiquitin-C (Val685) are post-translationally removed, followed by additional posttranslational cleavages to yield multiple copies of identical 76-amino acid ubiquitin, which is involved in one of the most common post-translational modifications of cellular proteins. Ub exists either covalently attached to another protein, or free (unanchored). Ub can be covalently linked to many cellular proteins by the ubiquitination process that targets proteins for degradation by the 26S proteasome. When covalently bound, it is conjugated to target proteins via an isopeptide bond either as a monomer (monoubiquitin), a polymer linked via different Lys residues of the ubiquitin (polyubiquitin chains) or a linear polymer linked via the initiator Met of the ubiquitin (linear polyubiquitin chains). Ub is linked covalently via its carboxyl terminus (Gly76) to usually lysine residues in target proteins. In a given target lysine residue can be linked to one single Ub molecule (monoubiquitylated) or to a chain of Ub molecules (polyubiquitylated). In a polyUb chain, Ub molecules can be linked through one of the seven lysines (K6, K11, K27, K29, K33, K48, and K63) or through the Ub N-terminus Met1 residue (which generates linear chains). Polyubiquitin chains, when attached to a target protein, have different functions depending on the lysine residue of the ubiquitin that is linked. Ub can be phosphorylated at serine 65 by PINK1 that leads to activation of the E3 ubiquitin ligase Parkin. PINK1 can depolarize mitochondria, which results in phosphorylation of Ub and activation of Parkin that can then induce mitophagy to eliminate damaged mitochondria. This ZooMAb^® recombinant monoclonal antibody, generated by our propriety technology, offers significantly enhanced specificity, affinity, reproducibility, and stability over conventional monoclonals.

Purified recombinant rabbit monoclonal antibody IgG, lyophilized in PBS with 5% Trehalose, normal appearance a coarse or translucent resin. Contains no biocide or preservatives, such as azide, or any animal by-products. Larger pack sizes provided as multiples of 25 μL.

0.03 mg/mL after reconstitution at 25 μL per vial. Please refer to guidance on suggested starting dilutions and/or titers per application and sample type.

Recommend storage of lyophilized product at 2-8°C; Before reconstitution, micro-centrifuge vials briefly to spin down material to bottom of the vial; Reconstitute each vial by adding 25 μL of filtered lab grade water or PBS; Reconstituted antibodies can be stored at 2-8°C, or -20°C for long term storage. Avoid repeated freeze-thaws.

ZooMAb is a registered trademark of Merck KGaA, Darmstadt, Germany

Unless otherwise stated in our catalog or other company documentation accompanying the product(s), our products are intended for research use only and are not to be used for any other purpose, which includes but is not limited to, unauthorized commercial uses, in vitro diagnostic uses, ex vivo or in vivo therapeutic uses or any type of consumption or application to humans or animals.