Revision 1
Cell Signaling Technology

Orders: 877-616-CELL (2355) [email protected]

Support: 877-678-TECH (8324)

Web: [email protected] cellsignal.com

3 Trask LaneDanversMassachusetts01923USA
For Research Use Only. Not for Use in Diagnostic Procedures.
Applications:

IF-IC, FC-FP

REACTIVITY:

H M

SENSITIVITY:

Endogenous

MW (kDa):

Source/Isotype:

Rabbit IgG

UniProt ID:

#P84243

Entrez-Gene Id:

3020

Product Information

Product Usage Information

Application Dilution
Immunofluorescence (Immunocytochemistry) 1:200
Flow Cytometry (Fixed/Permeabilized) 1:50

Storage

Supplied in PBS (pH 7.2), less than 0.1% sodium azide and 2 mg/ml BSA. Store at 4°C. Do not aliquot the antibody. Protect from light. Do not freeze.

Specificity / Sensitivity

Histone H3 (K27M Mutant Specific) (D3B5T) Rabbit mAb (Alexa Fluor® 488 Conjugate) recognizes endogenous levels of K27M mutant histone H3.1, H3.2, and H3.3 proteins. The antibody may cross-react with wild-type histone H3.1, 3.2, and 3.3 when used at a high concentration. Careful titration of this antibody may be required to obtain optimal specificity.

Species Reactivity:

Human, Mouse

Species predicted to react based on 100% sequence homology

Rat, Xenopus, Pig

Source / Purification

Monoclonal antibody is produced by immunizing animals with a synthetic peptide corresponding to K27M mutant sequence of human histone H3.3 protein.

Product Description

This Cell Signaling Technology antibody is conjugated to Alexa Fluor® 488 fluorescent dye and tested in-house for direct flow cytometric and immunofluorescent analysis in mouse cells and immunofluorescent analysis in human cells. This antibody is expected to exhibit the same species cross-reactivity as the unconjugated Histone H3 (K27M Mutant Specific) (D3B5T) Rabbit mAb #74829.

Background

Diffuse intrinsic pontine glioma (DIPG) is an aggressive brainstem astrocyte tumor arising mostly in children, leading to a long-term survival rate of less than 10%. Multiple whole-genome sequencing studies of DIPG patients identified commonly occurring mutations in the H3F3A gene encoding histone H3.3. One of these mutations, a lysine to methionine amino acid substitution (K27M), is found in up to 78% of DIPGs and 22% of non-brainstem pediatric gliomas (1-3). This mutation is associated with poor prognosis, with a mean survival time of 0.73 years for patients with the K27M mutation versus 4.6 years for patients without the mutation (1-3). Expression of the K27M mutant histone H3 is accompanied by a dramatic reduction in the levels of polycomb repressive complex 2 (PRC2)-mediated trimethylation of histone H3, changes in the distribution of PRC2 on the genome, and altered expression of genes associated with various cancer pathways (4-6).

  1. Wu, G. et al. (2012) Nat Genet 44, 251-3.
  2. Schwartzentruber, J. et al. (2012) Nature 482, 226-31.
  3. Khuong-Quang, D.A. et al. (2012) Acta Neuropathol 124, 439-47.
  4. Chan, K.M. et al. (2013) Genes Dev 27, 985-90.
  5. Lewis, P.W. et al. (2013) Science 340, 857-61.
  6. Piunti, A. et al. (2017) Nat Med 23, 493-500.

Species Reactivity

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

Applications Key

IF-IC: Immunofluorescence (Immunocytochemistry) FC-FP: Flow Cytometry (Fixed/Permeabilized)

Cross-Reactivity Key

H: human M: mouse R: rat Hm: hamster Mk: monkey Vir: virus Mi: mink C: chicken Dm: D. melanogaster X: Xenopus Z: zebrafish B: bovine Dg: dog Pg: pig Sc: S. cerevisiae Ce: C. elegans Hr: horse GP: Guinea Pig Rab: rabbit All: all species expected

Trademarks and Patents

Cell Signaling Technology is a trademark of Cell Signaling Technology, Inc.
XP is a registered trademark of Cell Signaling Technology, Inc.
This product is provided under an intellectual property license from Life Technologies Corporation. The transfer of this product is conditioned on the buyer using the purchased product solely in research conducted by the buyer, excluding contract research or any fee for service research, and the buyer must not (1) use this product or its components for (a) diagnostic, therapeutic or prophylactic purposes; (b) testing, analysis or screening services, or information in return for compensation on a per-test basis; or (c) manufacturing or quality assurance or quality control, and/or (2) sell or transfer this product or its components for resale, whether or not resold for use in research. For information on purchasing a license to this product for purposes other than as described above, contact Life Technologies Corporation, 5791 Van Allen Way, Carlsbad, CA 92008 USA or [email protected].
All other trademarks are the property of their respective owners. Visit cellsignal.com/trademarks for more information.

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Revision 1
#85023

Histone H3 (K27M Mutant Specific) (D3B5T) Rabbit mAb (Alexa Fluor® 488 Conjugate)

Immunofluorescence Image 1: Histone H3 (K27M Mutant Specific) (D3B5T) Rabbit mAb (Alexa Fluor® 488 Conjugate) Expand Image
Confocal immunofluorescent analysis of mouse astrocytes, containing either a knock-in FLAG-tagged K27M mutant histone H3.3 gene (left, positive) or a knock-in FLAG-tagged wild-type histone H3.3 gene (center, negative), and HeLa cells (right, negative), using Histone H3 (K27M Mutant Specific) (D3B5T) Rabbit mAb (Alexa Fluor® 488 Conjugate) (green) and β-Actin (8H10D10) Mouse mAb #3700 (red).
Flow Cytometry Image 1: Histone H3 (K27M Mutant Specific) (D3B5T) Rabbit mAb (Alexa Fluor® 488 Conjugate) Expand Image
Flow cytometric analysis of mouse astrocytes containing either a knock-in FLAG-tagged K27M mutant histone H3.3 gene (green) or a knock-in FLAG-tagged wild-type histone H3.3 gene (blue), using Histone H3 (K27M Mutant Specific) (D3B5T) Rabbit mAb (Alexa Fluor® 488 Conjugate) (solid lines) or a concentration-matched Rabbit (DA1E) mAb IgG XP® Isotype Control (Alexa Fluor® 488 Conjugate) #2975 (dashed lines).