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Polyubiquitin (K63-linkage-specific) monoclonal antibody (HWA4C4) (HRP conjugate)

BML-PW0605-0025 25 µg 155.00 USD
BML-PW0605-0100 100 µg 506.00 USD
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Product Specification

Immunogen:Synthetic peptide containing the ubiquitin K63-isopeptide motif.
UniProt ID:P0CG47 (UBB), P0CG48 (UBC)
Species reactivity:Species independent
Specificity:Recognizes K63-linked polyubiquitin in wide range of species.
Crossreactivity:Does not cross-react with any other isopeptide-linked (K6, K11, K27, K29, K33, or K48) polyubiquitinylated species or free ubiquitin.
Recommended Dilutions/Conditions:Western Blot (1:1,000)
Specificity for K63-isopeptide-linked polyubiquitin conjugates is temperature sensitive. Western blot membranes MUST NOT be autoclaved and excessive heat should avoided.
Suggested dilutions/conditions may not be available for all applications.
Optimal conditions must be determined individually for each application.
Application Notes:K63-linked polyubiquitin modified proteins comprise only a proportion of total cellular polyubiquitinylated species. Increased sample loading may therefore be required for analysis by Western blotting compared to equivalent experiments using general ubiquitin antibodies, despite the enhanced detection capability of the HWA4C4 HRP conjugated antibody.
Purity Detail:Protein G-affinity purified.
Formulation:Liquid. In 1X PBS, pH 7.2.
Handling:Avoid freeze/thaw cycles.
Shipping:Blue Ice Not Frozen
Short Term Storage:+4°C
Long Term Storage:-20°C
Scientific Background:Modification of proteins by addition of K63-linked polyubiquitin chains is implicated in a variety of cellular events, including DNA repair, signal transduction and receptor endocytosis.
bml-pw0605-pw8805 comp wb-s
Figure 1: Western blot of single isopeptide-linkage polyubiquitinylated conjugates (K6 - Prod. No. BML-UW0615, K11 - Prod. No. BML-UW0620, K27 - Prod. No. BML-UW0625, K29 - Prod. No. BML-UW0630, K33 - Prod. No. BML-UW0635, K48 - Prod. No. BML-UW0640, K63 - Prod. No. BML-UW0645, 1µg) using MAb to Polyubiquitinylated Conjugates (FK1) (Prod. No. BML-PW8805) and MAb to Polyubiquitin (K63-linkage-specific) (HWA4C4) (HRP) (Prod. No. BML-PW0605) at 1:1’000 dilution.
bml-pw0605 lysate-wb
Figure 2: Western blot of HeLa S100 cytosolic fraction (Prod. No. BML-SW8750, 5µg) probed with K63-linkage specific HRP-conjugate and general ubiquitin conjugate-reactive monoclonal antibodies at 1:1'000 dilution. Lane A. MAb to Polyubiquitin (K63-linkage-specific) (HWA4C4) (HRP) (Prod. No. BML-PW0605). Lane B. MAb to Mono- and Polyubiquitinylated Conjugates (FK2H) (HRP) (Prod. No. BML-PW0150).
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bml-pw0605-pw8805 comp wb-s bml-pw0605 lysate-wb

Product Literature References

S-nitrosylation-dependent proteasomal degradation restrains Cdk5 activity to regulate hippocampal synaptic strength: P. Zhang, et al.; Nat. Commun. 6, 8665 (2015), Application(s): Western blot, Abstract; Full Text

General Literature References

Analysis of nondegradative protein ubiquitylation with a monoclonal antibody specific for lysine-63-linked polyubiquitin: H. Wang, et al.; PNAS 105, 20197 (2008), Abstract;
Proteasome-independent functions of ubiquitin in endocytosis and signaling: D. Mukhopadhyay & H. Riezman; Science 315, 201 (2007), Abstract;
Distinct monoubiquitin signals in receptor endocytosis: K. Haglund, et al.; Trends Biochem. Sci. 28, 598 (2003), Abstract;
TAK1 is a ubiquitin-dependent kinase of MKK and IKK: C. Wang, et al.; Nature 412, 346 (2001), Abstract;
Cell cycle-regulated modification of the ribosome by a variant multiubiquitin chain: J. Spence, et al.; Cell 102, 67 (2000), Abstract;
Noncanonical MMS2-encoded ubiquitin-conjugating enzyme functions in assembly of novel polyubiquitin chains for DNA repair: R.M. Hofmann & C.M. Pickart; Cell 96, 645 (1999), Abstract;
A ubiquitin mutant with specific defects in DNA repair and multiubiquitination: J. Spence, et al.; Mol. Cell Biol. 15, 1265 (1995), Abstract;

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