Revision 1

#95132

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Orders:

877-616-CELL (2355)

[email protected]

Support:

877-678-TECH (8324)

3 Trask Lane | Danvers | Massachusetts | 01923 | USA

For Research Use Only. Not for Use in Diagnostic Procedures.

Applications:
W

Reactivity:
All

Sensitivity:
Endogenous

Source/Isotype:
Rabbit IgG

Product Usage Information

Application Dilution
Western Blotting 1:1000

Storage

Supplied in 10 mM sodium HEPES (pH 7.5), 150 mM NaCl, 100 µg/mL BSA, 50% glycerol, and less than 0.02% sodium azide. Store at –20°C. Do not aliquot the antibody.

Specificity/Sensitivity

beta-Hydroxybutyryl Lysine (E6H5Q) Rabbit Monoclonal Antibody recognizes a broad range of β-hydroxybutyryl lysine (Kbhb) containing proteins and peptides. This antibody does not cross-react with free β-hydroxybutyrate, and does not cross-react with peptides or proteins containing α-hydroxyisobutyrylated, acetylated, butyrylated, carboxymethylated, carboxyethylated, crotonylated, propionylated, or succinylated lysines.

Source / Purification

Monoclonal antibody is produced by immunizing animals with a synthetic peptide containing lysine β-hydroxybutyrylation flanked by degenerate amino acids at positions N- and C-terminal to the modified lysine.

Background

Lysine β-hydroxybutyrylation (Kbhb) is a reversible post-translational modification (PTM) that shares multiple regulatory enzymes with cellular acetylation machinery, including EP300 and HDAC1 (1). While acetylation is derived from acetyl-coenzyme-A (CoA), β-hydroxybutyrylation is derived from β-hydroxybutyryl-CoA, an intermediate generated from the metabolite β-hydroxybutyrate that is upregulated during nutrient limiting conditions such as starvation, ketogenic diets, and in diseases such as diabetes (2). The elucidation of the enzymes responsible for charging β-hydroxybutyrate with CoA to form β-hydroxybutyryl-CoA remains an active research area (3).

Notable bhb-modified substrates identified through proteomic investigations include histones, as well as non-histone substrates such as FASN and TP53 (4-6). In models where cells are incubated with excess β-hydroxybutyrate to induce this PTM, protein sites displaying upregulated bhb levels differ from sites displaying upregulated acetylation levels, suggesting potential differences in Kbhb-induced pathways and crosstalk between distinct modification types.  

Species Reactivity

Species reactivity is determined by testing in at least one approved application (e.g., western blot).

Western Blot Buffer

IMPORTANT: For western blots, incubate membrane with diluted primary antibody in 5% w/v BSA, 1X TBS, 0.1% Tween® 20 at 4°C with gentle shaking, overnight.

Applications Key

W: Western Blotting

Cross-Reactivity Key

All: All Species Expected

Trademarks and Patents

Cell Signaling Technology is a trademark of Cell Signaling Technology, Inc.

All other trademarks are the property of their respective owners. Visit cellsignal.com/trademarks for more information.

Limited Uses

Except as otherwise expressly agreed in a writing signed by a legally authorized representative of CST, the following terms apply to Products provided by CST, its affiliates or its distributors. Any Customer's terms and conditions that are in addition to, or different from, those contained herein, unless separately accepted in writing by a legally authorized representative of CST, are rejected and are of no force or effect.

Products are labeled with For Research Use Only or a similar labeling statement and have not been approved, cleared, or licensed by the FDA or other regulatory foreign or domestic entity, for any purpose. Customer shall not use any Product for any diagnostic or therapeutic purpose, or otherwise in any manner that conflicts with its labeling statement. Products sold or licensed by CST are provided for Customer as the end-user and solely for research and development uses. Any use of Product for diagnostic, prophylactic or therapeutic purposes, or any purchase of Product for resale (alone or as a component) or other commercial purpose, requires a separate license from CST. Customer shall (a) not sell, license, loan, donate or otherwise transfer or make available any Product to any third party, whether alone or in combination with other materials, or use the Products to manufacture any commercial products, (b) not copy, modify, reverse engineer, decompile, disassemble or otherwise attempt to discover the underlying structure or technology of the Products, or use the Products for the purpose of developing any products or services that would compete with CST products or services, (c) not alter or remove from the Products any trademarks, trade names, logos, patent or copyright notices or markings, (d) use the Products solely in accordance with CST Product Terms of Sale and any applicable documentation, and (e) comply with any license, terms of service or similar agreement with respect to any third party products or services used by Customer in connection with the Products.

Orders: 877-616-CELL (2355) [email protected] Support: 877-678-TECH (8324) [email protected] Web: cellsignal.com
For Research Use Only. Not for Use in Diagnostic Procedures.

Revision 1

Cell Signaling Technology Logo
Western blot analysis of extracts from NIH/3T3 and C6 cells, untreated (0 mM) or treated with increasing concentrations (10-80 mM) of sodium 3-hydroxybutyrate overnight, using β-Hydroxybutyryl Lysine (E6H5Q) Rabbit mAb (upper) or β-Actin (13E5) Rabbit mAb #4970 (lower) and detected with Anti-rabbit IgG (H+L) (DyLight 680 Conjugate) #5366.
Western Blotting Image 1: beta-Hydroxybutyryl Lysine (E6H5Q) Rabbit Monoclonal Antibody

Western blot analysis of extracts from HeLa cells, untreated (0 mM; -) or treated with sodium 3-hydroxybutyrate (80 mM; +) overnight, using β-Hydroxybutyryl Lysine (E6H5Q) Rabbit mAb or GAPDH (D16H11) XP® Rabbit mAb #5174 and detected with Anti-rabbit IgG (H+L) (DyLight 680 Conjugate) #5366. The specific β-hydroxybutyryl lysine (Kbhb) signal (first panel) was blocked by the modified peptide (third panel), but not by the corresponding unmodified peptide (second panel).

Western Blotting Image 2: beta-Hydroxybutyryl Lysine (E6H5Q) Rabbit Monoclonal Antibody
Orders: 877-616-CELL (2355) [email protected] Support: 877-678-TECH (8324) [email protected] Web: cellsignal.com
For Research Use Only. Not for Use in Diagnostic Procedures.