Histone H4 anticorps (3meLys20)

N° du produit ABIN7260402

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

Aperçu rapide pour Histone H4 anticorps (3meLys20) (ABIN7260402)

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)

Détail du produit anti-Histone H4 anticorps

Épitope: 3meLys20

Attributs du produit: Methylated antibody

Purification: Affinity purification

Immunogène: A synthetic methylated peptide corresponding to residues surrounding K20 of human histone H4

Isotype: IgG

Information d'application

Indications d'application: WB 1:500-1:2000 IHC 1:50-1:200 IF 1:50-1:200

Restrictions: For Research Use only

Independent Validation (IHC)

Validation #104614 (Immunohistochemistry)

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

No.: #104614

Date: 22.09.2024

Antigène: Histone H4

Numéro du lot: DL06BJD23422

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 (3meLys20) positive nuclei in the mouse brain. A: Immunoreactive nuclei in the granular layer of the cerebral cortex (original magnification 20x). There are several positive nuclei also in the white matter. Microwave antigen retrieval. B: Immunoreactive nuclei in the hippocampus. Note the intense staining of the CA neurons (original magnification 20x). Microwave antigen retrieval. C: Immunoreactive nuclei in the subventricular zone of the lateral ventriculus (original magnification 63x). Microwave antigen retrieval. Primary antibody dilution: 1:100.

Protocole

Anticorps primaire: ABIN7260401

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 (3meLys20) (ABIN7260401) 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 3x5 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 3x5 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 40x 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: PBS with 0.02 % sodium azide, 50 % glycerol, pH 7.3

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. Avoid freeze / thaw cycles.

Détails sur Histone H4

Antigène: Histone H4

Sujet: Histones are basic nuclear proteins that are responsible for the nucleosome structure of the chromosomal fiber in eukaryotes. This structure consists of approximately 146 bp of DNA wrapped around a nucleosome, an octamer composed of pairs of each of the four core histones (H2A, H2B, H3, and H4). The chromatin fiber is further compacted through the interaction of a linker histone, H1, with the DNA between the nucleosomes to form higher order chromatin structures. This gene is intronless and encodes a replication-dependent histone that is a member of the histone H4 family. Transcripts from this gene lack polyA tails, instead, they contain a palindromic termination element. This gene is found in a histone cluster on chromosome 1. This gene is one of four histone genes in the cluster that are duplicated, this record represents the centromeric copy.

Poids moléculaire:

Observed_MW: 11 kDa

Calculated_MW: 11 kDa

ID gène: 8370

UniProt: P62805