Histone H4 Antikörper (3meLys20)

Produktnummer ABIN7260402

Dieses Anti-Histone H4-Antikörper ist ein Kaninchen Polyklonal-Antikörper zur Detektion von Histone H4 in WB, IHC und IF. Geeignet für Human, Maus und Ratte. Unabhängig validiert für die Verwendung in Immunohistochemistry.

Kurzübersicht für Histone H4 Antikörper (3meLys20) (ABIN7260402)

Target: Histone H4

Reaktivität: Human, Maus, Ratte

Wirt: Kaninchen

Klonalität: Polyklonal

Konjugat: Dieser Histone H4 Antikörper ist unkonjugiert

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

Produktdetails anti-Histone H4 Antikörper

Bindungsspezifität: 3meLys20

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

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