Histone H4 antibody (acLys12)

Catalog No. ABIN6254899

Product Details anti-Histone H4 Antibody

Target: Histone H4

Binding Specificity: acLys12

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)

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

Abstract: Histone H4 Products

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