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

BML-PW0605

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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.

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Polyubiquitin (K63-linkage-specific) monoclonal antibody (HWA4C4) (HRP conjugate) Western blot — **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).”””””

K29 - Prod. No. BML-UW0630 — **Figure 1:** Western blot of single isopeptide-linkage polyubiquitinylated conjugates (K⁶ – Prod. No. BML-UW0615, K¹¹ – Prod. No. BML-UW0620

Product Details

Application	WB
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.
Clone	HWA4C4
Crossreactivity	Does not cross-react with any other isopeptide-linked (K6, K11, K27, K29, K33, or K48) polyubiquitinylated species or free ubiquitin.
Formulation	Liquid. In 1X PBS, pH 7.2.
Host	Mouse
Immunogen	Synthetic peptide containing the ubiquitin K63-isopeptide motif.
Isotype	IgG2a
Purity Detail	Protein G-affinity purified.
Recommendation 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.
Source	Purified from hybridoma tissue culture supernatant.
Species Reactivity	Species independent
Specificity	Recognizes K⁶³-linked polyubiquitin in wide range of species.
UniProt ID	P0CG47 (UBB), P0CG48 (UBC)
Worry-free Guarantee	This antibody is covered by our Worry-Free Guarantee.

Handling & Storage

Handling	Avoid freeze/thaw cycles.
Short Term Storage	+4°C
Long Term Storage	-20°C
Shipping	Blue Ice

Regulatory Status	RUO – Research Use Only

Method to Measure Ubiquitination of NLRs: Worboys, J. D., Palazón-Riquelme, P., et al.; Methods Mol. Biol. 2696, 105 (2023), Abstract
K48/K63-linked polyubiquitination of ATG9A by TRAF6 E3 ligase regulates oxidative stress-induced autophagy: Wang, Y. T., Liu, T. Y., et al.; Cell Rep. 38, 110354 (2022), Abstract
DNA damage and S phase-dependent E2F1 stabilization requires the cIAP1 E3-ubiquitin ligase and is associated with K63-poly-ubiquitination on lysine 161/164 residues.: Droin, N., Solary, E., et al.; Cell Death Dis. 8, e2816 (2017), Application(s): Ubiquitination assay / Reactant(s): Human, Abstract
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
Smad6 inhibits non-canonical TGF-β1 signalling by recruiting the deubiquitinase A20 to TRAF6.: Bae, Y. S., Oh, Y. S., et al.; Nat. Commun. 4, 2562 (2013), Application(s): WB / Reactant(s): Mouse, Abstract

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

BML-PW0605

Product Details

Handling & Storage

Datasheet, Manuals, SDS & CofA

Manuals And Inserts

Certificate of Analysis

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