Home Shop Polyubiquitin (K63-linkage-specific) monoclonal antibody (HWA4C4)

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

BML-PW0600

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Key antibody for the detection of K⁶³-linked polyubiquitinylated proteins.

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.

A monoclonal antibody (mAb) has been generated that specifically recognizes K63-linked polyubiquitin chains, but not any other isopeptide-linked (K⁶, K¹¹, K²⁷, K²⁹, K³³, or K⁴⁸) polyubiquitin chain or monoubiquitin. The specificity of this K⁶³-linked ubiquitinspecific monoclonal antibody was demonstrated using synthetic ubiquitin chains, single isopeptide-linked polyubiquitin chains, and with endogenously ubiquitinylated species in both cell lysates and intact cells.

The hybridoma secreting the antibody [clone HWA4C4] was generated by fusion of splenocytes from Balb/c mice which had received repeated immunisation with a synthetic peptide encompassing the ubiquitin K⁶³-isopeptide motif. Immunoglobulin was purified from tissue culture supernatant by ion exchange chromatography.

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This antibody is covered by our Worry-Free Guarantee.

Polyubiquitin (K63-linkage-specific) monoclonal antibody (HWA4C4) Immunohistochemistry — **Figure 3:** Formalin-fixed paraffin-embedded sections of hippocampus from human Alzheimer’s disease brain stained with MAb to Polyubiquitin (K⁶³-linkage-specific) (HWA4C4) at 1:100 dilution with biotinylated secondary antibody and DAB detection.

Polyubiquitin (K63-linkage-specific) monoclonal antibody (HWA4C4) Western blot — **Figure 2:** Western blot of HeLa S100 Fraction (BML-SW8750, 5µg) probed with MAb to Polyubiquitin (K⁶³-linkage-specific) (HWA4C4) (BML-PW0600) (Lane A) and MAb to Polyubiquitinylated Conjugates (FK1) (BML-PW8805) (Lane B) at 1:1’000 dilution.

Product Details

Application	ELISA, ICC, IHC, WB
Clone	HWA4C4
Crossreactivity	Does not cross-react with any other isopeptide-linked (K6, K11, K27, K29, K33, or K48) polyubiquitinylated species.
Formulation	Liquid. In PBS containing 0.09% sodium azide.
Host	Mouse
Immunogen	Synthetic peptide containing the ubiquitin K63-isopeptide motif.
Isotype	IgG2a
Recommendation Dilutions/Conditions	Immunocytochemistry (1:100)Immunohistochemistry (1:100)Western Blot (1:500 – 1:1,000)Specificity for K63 of mAb to Polyubiquitin (K63-linkage-specific) (HWA4C4) is temperature sensitive. Western blot membranes must not be autoclaved. Excessive heat should be avoided in all applications. 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 blot. Do not use milk as blocking solution. Use 1% BSA in an appropriate buffer instead.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. After opening, prepare aliquots and store at -20°C.
Long Term Storage	-20°C
Shipping	Blue Ice

Regulatory Status	RUO – Research Use Only

Targeted disruption of linkage-specific ubiquitylation reveals a key role of K29-linked ubiquitylation in epigenome integrity: Arroyo-Gomez, J., Wang, J., et al.; bioRxiv , (2024)
Stress granule homeostasis is modulated by TRIM21-mediated ubiquitination of G3BP1 and autophagy-dependent elimination of stress granules: Yang, C., Wang, Z., et al.; Autophagy 19, 1934 (2023), Abstract
NLRX1 ligand, docosahexaenoic acid, ameliorates LPS-induced inflammatory hyperalgesia by decreasing TRAF6/IKK/IkB-a/NF-kB signaling pathway activity: Yilmaz, D. E., Senol, S. P., et al.; Cell. Mol. Biol. (Noisy-le-grand) 69, 15 (2023), Abstract
Efficacy of IAPP suppression in mouse and human islets by GLP-1 analogue conjugated antisense oligonucleotide: Gurlo, T., Prakash, T. P., et al.; Front. Mol. Biosci. 10, 1096286 (2023), Abstract
Legionella longbeachae Regulates the Association of Polyubiquitinated Proteins on Bacterial Phagosome with Multiple Deubiquitinases: Y. Shi, et al.; Microbial. Spectr. 11, 4179 (2023), Abstract
Differential Ubiquitination as an Effective Strategy Employed by the Blood-Brain Barrier for Prevention of Bacterial Transcytosis: Bhutda, S., Ghosh, S., et al.; J. Bacteriol. 204, e0045621 (2022), Abstract
K27-linked ubiquitylation promotes p97 substrate processing and is essential for cell proliferation.: Shearer, R. F., Typas, D., et al.; EMBO J. 41, e110145 (2022), Reactant(s): Human, Abstract
MLN4924 Inhibits Defective Ribosomal Product Antigen Presentation Independently of Direct NEDDylation of Protein Antigens: K. Vijayasimha, et al.; J. Immunol. 208, 2273 (2022), Abstract
The SUMOylation of TAB2 mediated by TRIM60 inhibits MAPK/NF-κB activation and the innate immune response: Gu, Z., Chen, X., et al.; Cell. Mol. Immunol. 18, 1981 (2021), Abstract
Differential ubiquitination as an effective strategy employed by Blood-Brain Barrier for prevention of bacterial transcytosis: Ghosh, S., Banerjee, A., et al.; bioRxiv , (2021), Reactant(s): Human

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

BML-PW0600

Product Details

Handling & Storage

Datasheet, Manuals, SDS & CofA

Manuals And Inserts

Certificate of Analysis

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