Render Target: STATIC
Render Timestamp: 2025-01-22T05:16:10.762Z
Commit: da7e4f2f0d1aed1f1f8e20e4e2ecab8f33cbd595
XML generation date: 2024-12-25 01:36:47.788
Product last modified at: 2025-01-01T09:02:21.970Z
1% for the planet logo
PDP - Template Name: ELISA Kit
PDP - Template ID: *******bd382c2

PathScan® Total Histone H3 Sandwich ELISA Kit #7253

Filter:
  • ELISA
ELISA Image 1: PathScan® Total Histone H3 Sandwich ELISA Kit
Figure 1. Treatment of NIH/3T3 cells with trichostatin A (TSA) increases the acetylation of histone H3 at Lys9, detected by PathScan® Acetyl-Histone H3 (Lys9) Sandwich ELISA Kit #7121, and the di-methylation of histone H3 at Lys4, detected by PathScan® Di-Methyl-Histone H3 (Lys4) Sandwich ELISA Kit #7124. However, TSA treatment does not affect the level of total histone H3 that is detected by Pathscan® Total Histone H3 Sandwich ELISA Kit #7253. NIH/3T3 cells (70-80% confluent) were treated for 16-18 hours with 0.4 μM TSA at 37ºC. Absorbance readings at 450 nm are shown in the top figure while the corresponding Western blots using Histone H3 Antibody #9715 (left panel), Acetyl-Histone H3 (Lys9) Antibody #9671 (middle panel) or Di-Methyl-Histone H3 (Lys4) (C64G9) Rabbit mAb #9725 (right panel) are shown in the bottom figure.

To Purchase # 7253

Cat. #

Size

7253C
1 Kit
7253V1
5 Kits
7253V2
10 Kits
7253V3
20 Kits
7253V4
50 Kits

Important Ordering Details

Custom Ordering Details:

If kit quantities from the same lot are needed in unlisted sizes, contact us for processing time and pricing.

Looking for this ELISA kit in a 384-well format? Inquire for availability, processing time, and pricing.

Supporting Data

REACTIVITY H M Mk
Application Key:
  • ELISA-ELISA 
Species Cross-Reactivity Key:
  • H-Human 
  • M-Mouse 
  • Mk-Monkey 
  • Product Includes
  • Related Products
Product IncludesVolumeSolution Color
Histone H3 Rabbit mAb Coated Microwells #3300296 tests
Histone H3 Mouse Detection mAb #142211 eaGreen (Lyophilized)
Anti-mouse IgG, HRP-linked Antibody (ELISA Formulated) #133041 eaRed (Lyophilized)
Detection Antibody Diluent #1333911 mlGreen
HRP Diluent #1351511 mlRed
TMB Substrate #700411 ml
STOP Solution #700211 ml
Sealing Tape #545032 ea
ELISA Wash Buffer (20X) #980125 ml
ELISA Sample Diluent #1108325 mlBlue
Cell Lysis Buffer (10X) #980315 ml

Kit contents scale proportionally with size, except sealing tape.
Example: The V1 kit contains 5X the listed quantities above, but will exclude the sealing tape.

The microwell plate is supplied as 12 8-well modules - Each module is designed to break apart for 8 tests.

Product Information

Product Description

The PathScan® Total Histone H3 Sandwich ELISA Kit is a solid phase sandwich enzyme-linked immunosorbent assay (ELISA) that detects endogenous levels of histone H3. A histone H3 rabbit antibody has been coated onto the microwells. After incubation with cell lysates, histone H3 (modified and unmodified) is captured by the coated antibody. Following extensive washing, histone H3 mouse mAb is added to detect the captured histone H3 protein. Anti-Mouse IgG, HRP-linked antibody is then used to recognize the bound detection antibody. HRP substrate, TMB, is added to develop color. The magnitude of the absorbance for this developed color is proportional to the quantity of total histone H3.

*Antibodies in this kit are custom formulations specific to kit.

Protocol

Specificity / Sensitivity

CST's PathScan® Total Histone H3 Sandwich ELISA Kit #7253 detects endogenous levels of histone H3. As shown in Figure 1, using the Total Histone H3 Sandwich ELISA Kit #7253, a high level of histone H3 is detected in NIH/3T3 cells when treated with TSA. The level of total histone H3 (modified and unmodified) remains unchanged as shown by Western analysis (Figure 1). Similar results are obtained when COS and Jurkat cells are treated with TSA (data not shown). Note: For this assay, it is recommended that lysates be thoroughly sonicated to ensure complete extraction of histone H3 and an accurate absorbance reading. This kit detects proteins from the indicated species, as determined through in-house testing, but may also detect homologous proteins from other species.

Species Reactivity:

Human, Mouse, Monkey

Background

Modulation of chromatin structure plays an important role in the regulation of transcription in eukaryotes. The nucleosome, made up of DNA wound around eight core histone proteins (two each of H2A, H2B, H3, and H4), is the primary building block of chromatin (1). The amino-terminal tails of core histones undergo various posttranslational modifications, including acetylation, phosphorylation, methylation, and ubiquitination (2-5). These modifications occur in response to various stimuli and have a direct effect on the accessibility of chromatin to transcription factors and, therefore, gene expression (6). In most species, histone H2B is primarily acetylated at Lys5, 12, 15, and 20 (4,7). Histone H3 is primarily acetylated at Lys9, 14, 18, 23, 27, and 56. Acetylation of H3 at Lys9 appears to have a dominant role in histone deposition and chromatin assembly in some organisms (2,3). Phosphorylation at Ser10, Ser28, and Thr11 of histone H3 is tightly correlated with chromosome condensation during both mitosis and meiosis (8-10). Phosphorylation at Thr3 of histone H3 is highly conserved among many species and is catalyzed by the kinase haspin. Immunostaining with phospho-specific antibodies in mammalian cells reveals mitotic phosphorylation at Thr3 of H3 in prophase and its dephosphorylation during anaphase (11).

Pathways

Explore pathways related to this product.


For Research Use Only. Not For Use In Diagnostic Procedures.
Cell Signaling Technology is a trademark of Cell Signaling Technology, Inc.
PathScan is a registered trademark of Cell Signaling Technology, Inc.
All other trademarks are the property of their respective owners. Visit our Trademark Information page.