Revision 1

#65200

Store at -20C

CST Logo
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:
SCA-ITS3

Reactivity:
H M

Sensitivity:
Endogenous

Source/Isotype:
Rabbit IgG

UniProt ID:
#P68431

Entrez-Gene Id:
8350

Storage

Supplied in PBS (pH 7.2), 2 mM EDTA, 0.05% Triton X-100, 2 mg/mL BSA, and 50% glycerol. Store at -20°C. Do not aliquot the antibody.

Specificity/Sensitivity

Tri-Methyl-Histone H3 (Lys4) (C42D8) Rabbit Monoclonal Antibody (InTraSeq™ 3' Conjugate 3085) detects endogenous levels of histone H3 when tri-methylated on Lys4. This antibody conjugate shows some cross-reactivity with histone H3 that is di-methylated on Lys4, but does not cross-react with non-methylated or mono-methylated histone H3 Lys4. In addition, this antibody conjugate does not cross-react with methylated histone H3 Lys9, Lys27, Lys36 or methylated histone H4 Lys20.

Source / Purification

Monoclonal antibody is produced by immunizing animals with a synthetic peptide corresponding to the amino terminus of histone H3 in which Lys4 is tri-methylated.

Background

The nucleosome, made up of four core histone proteins (H2A, H2B, H3, and H4), is the primary building block of chromatin. Originally thought to function as a static scaffold for DNA packaging, histones have now been shown to be dynamic proteins, undergoing multiple types of post-translational modifications, including acetylation, phosphorylation, methylation, and ubiquitination (1). Histone methylation is a major determinant for the formation of active and inactive regions of the genome and is crucial for the proper programming of the genome during development (2,3). Arginine methylation of histones H3 (Arg2, 17, 26) and H4 (Arg3) promotes transcriptional activation and is mediated by a family of protein arginine methyltransferases (PRMTs), including the co-activators PRMT1 and CARM1 (PRMT4) (4). In contrast, a more diverse set of histone lysine methyltransferases has been identified, all but one of which contain a conserved catalytic SET domain originally identified in the Drosophila Su(var)3-9, Enhancer of zeste, and Trithorax proteins. Lysine methylation occurs primarily on histones H3 (Lys4, 9, 27, 36, 79) and H4 (Lys20) and has been implicated in both transcriptional activation and silencing (4). Methylation of these lysine residues coordinates the recruitment of chromatin modifying enzymes containing methyl-lysine binding modules such as chromodomains (HP1, PRC1), PHD fingers (BPTF, ING2), tudor domains (53BP1), and WD-40 domains (WDR5) (5-8). The discovery of histone demethylases, such as PADI4, LSD1, JMJD1, JMJD2, and JHDM1, has shown that methylation is a reversible epigenetic marker (9).

Species Reactivity

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

Applications Key

SCA-ITS3: Single Cell Analysis - InTraSeq™ 3'

Cross-Reactivity Key

H: Human M: Mouse

Trademarks and Patents

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

10x Genomics, 10x, Feature Barcode, and Chromium are the trademarks or registered trademarks of 10x Genomics, Inc.

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

InTraSeq is a trademark of Cell Signaling Technology, Inc.

Subject to patents licensed from 10x Genomics, Inc. for use with single-cell (i.e., Chromium) 10x products.

U.S. Patent No. 7,429,487, foreign equivalents, and child patents deriving therefrom.

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
HeLa cells, HeLa cells treated with UV (100 mJ with 2 hr recovery), Jurkat cells, and SCLC21H cells were processed in accordance with the InTraSeq™ 3’ protocol and analyzed using the Seurat computational package. The left figure shows the Uniform Manifold Approximation and Projection (UMAP) of the annotated cells. The FeaturePlot in the right figure displays the Tri-Methyl-Histone H3 (Lys4) (C42D8) Rabbit mAb (InTraSeq™ 3' Conjugate 3085) #65200 expression. To generate similar plots, use the FeaturePlot command template:
FeaturePlot(your_object, features = your_protein, max.cutoff = "q95", min.cutoff = "q5") + scale_color_gradientn(colours = c("#22578B", "#73A9D2", "grey90", "#FDB393", "#b22222"))

SCA Image 1: Tri-Methyl-Histone H3 (Lys4) (C42D8) Rabbit Monoclonal Antibody (InTraSeq<sup>™</sup> 3' Conjugate 3085)
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.