Histone H4 anticorps (acLys12)

N° du produit ABIN6254899

Cet anticorps anti-Histone H4 est un anticorps Lapin Polyclonal détectant Histone H4 dans WB, IHC, IF, ELISA et ICC. Adapté pour Humain, Souris et Rat. Validé indépendamment pour une utilisation dans Immunohistochemistry.

Aperçu rapide pour Histone H4 anticorps (acLys12) (ABIN6254899)

Antigène: Histone H4

Reactivité: Humain, Souris, Rat

Hôte: Lapin

Clonalité: Polyclonal

Conjugué: Cet anticorp Histone H4 est non-conjugé

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

Détail du produit anti-Histone H4 anticorps

Épitope: acLys12

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

Réactivité croisée: Humain, Souris, Rat (Rattus)

Purification: affinity purification

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

Isotype: IgG

Information d'application

Indications d'application: 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: 22.09.2024

Antigène: Histone H4

Numéro du lot: lk4553

Application validée: Immunohistochemistry

Contrôle positif:

CD1 mouse brain fixed in 4% paraformaldehyde.

Contrôle négative:

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

Protocole

Anticorps primaire: ABIN6254899

Anticorps secondaire: 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.

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.

Stockage

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.

Agent conservateur: Sodium azide

Précaution d'utilisation: This product contains Sodium azide: a POISONOUS AND HAZARDOUS SUBSTANCE which should be handled by trained staff only.

Stock: -20 °C

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

Date de péremption: 12 months

Détails sur Histone H4

Antigène: Histone H4

Sujet:

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

Poids moléculaire: 11kDa

ID gène: 121, 504, 554

UniProt: P62805