Histone H4 Antikörper (3meLys20)

Produktnummer ABIN7260402

Produktdetails anti-Histone H4 Antikörper

Target: Histone H4

Bindungsspezifität: 3meLys20

Reaktivität: Human, Maus, Ratte

Wirt: Kaninchen

Klonalität: Polyklonal

Konjugat: Dieser Histone H4 Antikörper ist unkonjugiert

Applikation: Western Blotting (WB), Immunohistochemistry (IHC), Immunofluorescence (IF)

Produktmerkmale: Methylated antibody

Aufreinigung: Affinity purification

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

Isotyp: IgG

Anwendungsinformationen

Applikationshinweise: WB 1:500-1:2000 IHC 1:50-1:200 IF 1:50-1:200

Beschränkungen: Nur für Forschungszwecke einsetzbar

Independent Validation (IHC)

Validierung #104614 (Immunohistochemistry)

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

No.: #104614

Datum: 22.09.2024

Antigen: Histone H4

Chargennummer: DL06BJD23422

Validierte Anwendung: Immunohistochemistry

Positivkontrolle:

CD1 mouse brain fixed in 4% paraformaldehyde.

Negativkontrolle:

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

Validierungsbilder

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.

Protokoll

Primärantikörper: ABIN7260401

Sekundärantikörper: 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.

Anmerkungen:

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.

Handhabung

Format: Liquid

Konzentration: 1 mg/mL

Buffer: PBS with 0.02 % sodium azide, 50 % glycerol, pH 7.3

Konservierungsmittel: Sodium azide

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

Lagerung: -20 °C

Informationen zur Lagerung: Store at -20°C. Avoid freeze / thaw cycles.

Details zu Histone H4

Target: Histone H4

Abstract: Histone H4 Produkte

Hintergrund: 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.

Molekulargewicht:

Observed_MW: 11 kDa

Calculated_MW: 11 kDa

Gen-ID: 8370

UniProt: P62805