Histone H4 antibody (acLys12)

Catalog No. ABIN6254899

This anti-Histone H4 antibody is a Rabbit Polyclonal antibody detecting Histone H4 in WB, IHC, IF, ELISA and ICC. Suitable for Human, Mouse and Rat. Independently validated for use in Immunohistochemistry.

Quick Overview for Histone H4 antibody (acLys12) (ABIN6254899)

Target: Histone H4

Reactivity: Human, Mouse, Rat

Host: Rabbit

Clonality: Polyclonal

Conjugate: This Histone H4 antibody is un-conjugated

Application: Western Blotting (WB), Immunohistochemistry (IHC), Immunofluorescence (IF), ELISA, Immunocytochemistry (ICC)

Product Details anti-Histone H4 Antibody

Binding Specificity: acLys12

Specificity: Acetyl-Histone H4 (Lys12) Antibody detects endogenous levels of total Histone H4 protein only when acetylated at lysine12.

Cross-Reactivity: Human, Mouse (Murine), Rat (Rattus)

Purification: affinity purification

Immunogen: The antiserum was produced against synthesized peptide derived from human Histone H4 around the acetylated site of Lys12.

Isotype: IgG

Application Details

Application Notes: WB: 1:500~1:2000 IHC: 1:50~1:200 IF 1:200

Restrictions: For Research Use only

Independent Validation (IHC)

Validation #104615 (Immunohistochemistry)

by: Prof. Merighi, Laboratory of Neurobiology, Department of Veterinary Sciences, University of Turin

No.: #104615

Date: 09/22/2024

Antigen: Histone H4

Lot Number: lk4553

Method validated: Immunohistochemistry

Positive Control:

CD1 mouse brain fixed in 4% paraformaldehyde.

Negative Control:

Control slices were processed for each experimental procedure, omitting the primary antibody. In this case, overnight incubation was only performed using a diluent solution.

Validation Images

Histone H4 (acLys12) positive nuclei in the mouse brain. A: Immunoreactive nuclei in the granular layer of the cerebral cortex (original magnification 20x). Microwave antigen retrieval. B: Immunoreactive nuclei in the hippocampus. Note the intense staining

Full Methods

Primary Antibody: ABIN6254899

Secondary Antibody: Alexa Fluor 488 goat anti-rabbit (Invitrogen by Thermo Fisher Scientific, A11034, lot # 2380031), Alexa Fluor 488 donkey anti-rabbit (Invitrogen by Thermo Fisher Scientific A32790 lot # XJ357262).

Full Protocol:

• SAMPLE ANALYZED: Old (> 18 months) CD1 mouse brain.
• Mice were perfused with paraformaldehyde 4% in 0.1M phosphate buffer (PB) pH 7.4 and samples were post-fixed in the same fixative for 2 additional hours at room temperature. After fixation, samples were washed, dehydrated, and embedded in paraffin wax.
• Following several washes in PBS, the brain was cut with a microtome (6 μm-thick sections) in parasagittal sections mounted on glass slides.
• After paraffin removal, sections were incubated for 1 hour at room temperature in PBS containing 1% albumin from chicken egg white (Sigma, A5378) and 0.3% Triton-X-100 (BioRad, 161-0407, lot # 00583) to block non-specific binding sites. Sections were then incubated overnight at room temperature with the primary antibody anti-Histone H4 (acLys12) (ABIN6254899) at 1:50, 1:100, and 1:200, in PBS-BSA (Sigma, A7906)-PLL (Sigma, P1524).
• Antigen retrieval treatment was also tested. In this case, sections were processed for microwave antigen retrieval for 10 minutes (95–100°C) in 10 mM sodium citrate buffer (pH 6.0). After 20 minutes of spontaneous cooling, sections were washed (distilled water, 2 times for 5 min each; followed by PBS for 5 min).
• Sections were washed 3×5 min in 0.01M PBS.
• Sections were then incubated with the anti-rabbit secondary antibody Alexa Fluor 488, 1:500, in 0.1M PBS, for 1 hour at room temperature.
• Sections were washed 3×5 min in 0.01M PBS.
• Specimens were mounted in Fluoroshield (Sigma, F6182, lot # MKCB0153V).
• Images were acquired with a Leica DM 6000B fluorescence microscope with a digital camera at 20× or 40× magnification.

Experimental Notes:

Passed. The antibody works for IHC-P at 1:100 and 1:200 concentrations with or without microwave antigen retrieval. Microwave treatment increases the intensity of staining.

Handling

Format: Liquid

Concentration: 1 mg/mL

Buffer: Rabbit IgG in phosphate buffered saline ,  pH 7.4, 150  mM NaCl, 0.02 % sodium azide and 50 % glycerol.

Preservative: Sodium azide

Precaution of Use: This product contains Sodium azide: a POISONOUS AND HAZARDOUS SUBSTANCE which should be handled by trained staff only.

Storage: -20 °C

Storage Comment: Store at -20 °C.Stable for 12 months from date of receipt

Expiry Date: 12 months

Target Details for Histone H4

Target: Histone H4

Background:

Description: Core component of nucleosome. Nucleosomes wrap and compact DNA into chromatin, limiting DNA accessibility to the cellular machineries which require DNA as a template. Histones thereby play a central role in transcription regulation, DNA repair, DNA replication and chromosomal stability. DNA accessibility is regulated via a complex set of post-translational modifications of histones, also called histone code, and nucleosome remodeling.

Gene: HIST1H4A, HIST1H4B, HIST1H4C, HIST1H4D, HIST1H4E, HIST1H4F, HIST1H4H, HIST1H4I, HIST1H4J, HIST1H4K, HIST1H4L, HIST2H4A, HIST2H4B, HIST4H4

Molecular Weight: 11kDa

Gene ID: 121, 504, 554

UniProt: P62805