ORC1 Antikörper

Produktnummer ABIN7252684

Der Kaninchen Polyklonal anti-ORC1 Antikörper wird verwendet zum Nachweis von ORC1 in Proben von Human, Maus und Ratte. Er wurde validiert für ELISA und IHC. Der Antikörper wurde zudem unabhängig validiert.

Kurzübersicht für ORC1 Antikörper (ABIN7252684)

Target: ORC1 (ORC1L) (Origin Recognition Complex, Subunit 1 (ORC1L))

Reaktivität: Human, Maus, Ratte

Wirt: Kaninchen

Klonalität: Polyklonal

Konjugat: Dieser ORC1 Antikörper ist unkonjugiert

Applikation: ELISA, Immunohistochemistry (IHC)

Produktdetails anti-ORC1 Antikörper

Produktmerkmale: Polyclonal Antibody

Aufreinigung: Antigen affinity purification

Immunogen: Fusion protein of human ORC1

Isotyp: IgG

Anwendungsinformationen

Applikationshinweise: IHC 1:50-1:300, ELISA 1:5000-1:10000

Beschränkungen: Nur für Forschungszwecke einsetzbar

Independent Validation (IHC)

Validierung #104637 (Immunohistochemistry)

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

No.: #104637

Datum: 18.03.2026

Antigen: Origin Recognition Complex, Subunit 1 (ORC1)

Chargennummer: DW04006249610

Validierte Anwendung: Immunohistochemistry

Positivkontrolle:

CD1 mouse brain fixed in 4% paraformaldehyde.

Negativkontrolle:

Omission of primary/secondary antibody.

Bewertung:

The antibody works in IHC-P and IHC (fro) at concentrations from 1:50-1:200. Microwave pretreatment is strongly suggested.

Validierungsbilder

Origin Recognition Complex, Subunit 1 (ORC1L) immunoreactive cells in the cerebral cortex and hippocampus of P5 and adult CD1 mice. IHC-P. Magnification: 63x.

Protokoll

Primärantikörper: ABIN7252683

Sekundärantikörper: anti-rabbit Alexa Fluor 488 (Invitrogen A11008, lot # 3112790)

Full Protocol:

• IHC (p)
  • PROTOCOL
    • Sample analyzed: P5 and adult (9 months old) CD1 mouse brain
    • Paraffin-embedded slices (7µm thick) were deparaffinized and rehydrated through a graded series of alcohols and distilled water (dH2O)
    • Sections were blocked in ovalbumin 1% 1 hour at room temperature
    • Antigen retrieval was performed in all the sections by microwave treatment (1 minute at 750W + 1 minute at 250W in Citrate buffer pH 6)
    • 3x5 minutes washing in 0.01M PBS
    • Sections were incubated with the primary antibody at the following dilutions of 1:50/1:100/1:200, overnight at room temperature (primary antibody was diluted in PLL/BSA/PBS diluent)
    • 3x5 minutes washing in 0.01M PBS
    • Incubation with the anti-rabbit secondary antibody 1:400 in PLL/BSA/PBS diluent, 1 hour at room temperature.
    • 3x5 minutes washing in 0.01M PBS
    • Nuclear counterstaining was performed with 4′,6-Diamidine-2′-phenylindole dihydrochloride (DAPI) 1µg/mL, 2 minutes
    • 2x5 minutes washing in 0.01M PBS
    • Specimens were then mounted in Fluoroshield mounting medium (Sigma)
    • Images were acquired with a Leica DM 6000B fluorescence microscope with a digital camera at 63x magnification
• IHC (fro)
  • PROTOCOL
    • Sample analyzed: old (22 months old) CD1 mouse brain
    • Samples were fixed in paraformaldehyde and cryoprotected with a series of sucrose solutions in 0.01M PBS at increasing concentrations (10%, 20%, 30%), then embedded with OCT compound in isopentane previously cooled in liquid nitrogen
    • Frozen slices (10µm thick) were washed 3x5 minutes in 0.01M PBS
    • Sections were blocked in ovalbumin 1% 1 hour at room temperature
    • Antigen retrieval was performed in all the sections by microwave treatment (1 minute at 750W + 1 minute at 250W in Citrate buffer pH 6)
    • 3x5 minutes washing in 0.01M PBS
    • Sections were incubated with the primary antibody at the following dilutions of 1:50/1:100/1:200, overnight at room temperature (primary antibody was diluted in PLL/BSA/PBS diluent)
    • 3x5 minutes washing in 0.01M PBS
    • Incubation with the anti-rabbit secondary antibody 1:400 in PLL/BSA/PBS diluent, 1 hour at room temperature.
    • 3x5 minutes washing in 0.01M PBS
    • Nuclear counterstaining was performed with 4′,6-Diamidine-2′-phenylindole dihydrochloride (DAPI) 1µg/mL, 2 minutes
    • 2x5 minutes washing in 0.01M PBS
    • Specimens were then mounted in Fluoroshield mounting medium (Sigma)
    • Slides were observed under a Leica DM 6000B fluorescence microscope

Anmerkungen:

• For both IHC-P and IHC (fro) some slides were processed without microwave treatment to evaluate the actual necessity of antigen retrieval.
• To exclude any nonspecific signal due to the secondary antibody, one control slice was processed for each experimental procedure, omitting the primary antibody.

Handhabung

Format: Liquid

Konzentration: 1.14 mg/mL

Buffer: PBS with 0.05 % Sodium azide and 40 % Glycerol, pH 7.4

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 ORC1

Target: ORC1 (ORC1L) (Origin Recognition Complex, Subunit 1 (ORC1L))

Andere Bezeichnung: ORC1

Hintergrund: The origin recognition complex (ORC) is a highly conserved six subunits protein complex essential for the initiation of the DNA replication in eukaryotic cells. Studies in yeast demonstrated that ORC binds specifically to origins of replication and serves as a platform for the assembly of additional initiation factors such as Cdc6 and Mcm proteins. The protein encoded by this gene is the largest subunit of the ORC complex. While other ORC subunits are stable throughout the cell cycle, the levels of this protein vary during the cell cycle, which has been shown to be controlled by ubiquitin-mediated proteolysis after initiation of DNA replication. This protein is found to be selectively phosphorylated during mitosis. It is also reported to interact with MYST histone acetyltransferase 2 (MyST2/HBO1), a protein involved in control of transcription silencing. Alternatively spliced transcript variants encoding different isoforms have been found for this gene.ORC1 (Origin Recognition Complex Subunit 1) is a Protein Coding gene. Diseases associated with ORC1 include Meier-Gorlin Syndrome 1 and Microtia. Among its related pathways are E2F mediated regulation of DNA replication and CDK-mediated phosphorylation and removal of Cdc6. GO annotations related to this gene include chromatin binding. An important paralog of this gene is CDC6.

UniProt: Q13415

Pathways: Mitotic G1-G1/S Phases, DNA Replication, Synthesis of DNA