Histone H4 Antikörper (acLys12)

Produktnummer ABIN6254899

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

Kurzübersicht für Histone H4 Antikörper (acLys12) (ABIN6254899)

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), ELISA, Immunocytochemistry (ICC)

Produktdetails anti-Histone H4 Antikörper

Bindungsspezifität: acLys12

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

Kreuzreaktivität: Human, Maus, Ratte (Rattus)

Aufreinigung: affinity purification

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

Isotyp: IgG

Anwendungsinformationen

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

Beschränkungen: Nur für Forschungszwecke einsetzbar

Independent Validation (IHC)

Validierung #104615 (Immunohistochemistry)

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

No.: #104615

Datum: 22.09.2024

Antigen: Histone H4

Chargennummer: lk4553

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 (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

Protokoll

Primärantikörper: ABIN6254899

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 (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.

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: Rabbit IgG in phosphate buffered saline ,  pH 7.4, 150  mM NaCl, 0.02 % sodium azide and 50 % glycerol.

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.Stable for 12 months from date of receipt

Haltbarkeit: 12 months

Details zu Histone H4

Target: Histone H4

Hintergrund:

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

Molekulargewicht: 11kDa

Gen-ID: 121, 504, 554

UniProt: P62805