MYCN Antikörper (AA 401-464)

Produktnummer ABIN760676

Der Kaninchen Polyklonal Anti-MYCN-Antikörper wurde für ELISA, IF (cc), IF (p), IHC (p) und IHC (fro) validiert und wurde unabhängig validiert. Er ist geeignet, MYCN in Proben von Human und Maus zu detektieren. Es ist 1 Publikation verfügbar.

Kurzübersicht für MYCN Antikörper (AA 401-464) (ABIN760676)

Target: MYCN (N-Myc Proto-Oncogene Protein (MYCN))

Reaktivität: Human, Maus

Wirt: Kaninchen

Klonalität: Polyklonal

Konjugat: Dieser MYCN Antikörper ist unkonjugiert

Applikation: ELISA, Immunofluorescence (Cultured Cells) (IF (cc)), Immunofluorescence (Paraffin-embedded Sections) (IF (p)), Immunohistochemistry (Paraffin-embedded Sections) (IHC (p)), Immunohistochemistry (Frozen Sections) (IHC (fro))

Produktdetails anti-MYCN Antikörper

Bindungsspezifität: AA 401-464

Homologie: Rat,Cow,Pig

Aufreinigung: Purified by Protein A.

Immunogen: KLH conjugated synthetic peptide derived from human n-Myc

Isotyp: IgG

Anwendungsinformationen

Applikationshinweise: ELISA 1:500-1000
IHC-P 1:200-400
IHC-F 1:100-500
IF(IHC-P) 1:50-200
IF(IHC-F) 1:50-200
IF(ICC) 1:50-200

Beschränkungen: Nur für Forschungszwecke einsetzbar

Independent Validation (WB)

Validierung #029800 (Western Blotting)

by: Alamo Laboratories Inc

No.: #029800

Datum: 21.08.2014

Chargennummer: 120917

Validierte Anwendung: Western Blotting

Positivkontrolle: MCF7 cells (variant MCF7/VP)00284-0/pdf)

Negativkontrolle: SK-N-SH cells - RNA

Bewertung: A strong band was observed in the positive control sample at the correct molecular weight. An additional band was also observed in the positive sample at a higher molecular weight which may represent non-specific binding. No bands were observed in the negative control sample.

Validierungsbilder

Figure 1: Western Blot for MYCN. Arrowhead indicates the expected molecular weight of ~49 kDa.

Protokoll

Primärantikörper:

• Antigen: Mycn (MYCN)
• Catalog number: ABIN760676
• Lot number: 120917
• Antibody Dilution: 1:200

Sekundärantikörper:

• Antigen: Goat Anti-Rabbit IgG (H + L)-HRP Conjugate
• Lot number: L170-6515
• Antibody Dilution: 1:10,000

Full Protocol:

• 1. The cell extracts were heated at 95°C for 5 minutes in 1X SDS Sample Buffer containing 1% SDS and 1.25% β-mercaptoethanol.
• 2. 15 μl of heated culture-media were loaded and resolved on 8-16% SDS-polyacrylamide gel.
• 3. The Thermo Scientific - Spectra Multicolor Broad Range (Cat # 26634) were used as molecular mass markers.
• 4. Proteins were then transferred onto PVDF membrane by wet transfer and protein transfer was confirmed with Ponceau-S staining.
• 5. The PVDF membrane was incubated with 25 ml of blocking buffer [Tris Buffered Saline, pH 7.4 plus 0.1% TW20 (TBST)] containing 5% (W/V) BSA at room temperature for 1 hour.
• 6. The membrane was rinsed with TBST once.
• 7. The membrane was immersed with the protein side up in the primary antibody solution in TBST containing 5% (W/V) BSA and incubated for 13 hours at 4°C.
• 8. The membrane was rinsed in TBST thrice for 5 minutes each.
• 9. The membrane was incubated in the HRP-conjugated secondary antibody solution in TBST containing 5% (W/V) BSA and incubated for 1 hour at room temperature (~26°C) with gentle agitation.
• 10. The membrane was rinsed thrice TBST thrice for 5 minutes each.
• 11. The membrane was rinsed in TBS twice for 30 seconds each.
• 12. Signals were detected with ECL-2 Substrate. The blot was scanned for 45 minutes.
• 13. The membrane was rinsed three times TBST.
• 14. Incubated in Acidic Glycine Stripping Buffer at room temperature with gentle agitation for 3 times, 10 minutes each.
• 15. The membrane was washed in TBST 2 times for 10 minutes each.
• 16. Repeated Steps 5-12 with the loading control antibody (for Anti-actin) and its matching secondary antibody.

Anmerkungen: - No experimental challenges noted.

Handhabung

Format: Liquid

Konzentration: 1 μg/μL

Buffer: 0.01M TBS( pH 7.4) with 1 % BSA, 0.03 % Proclin300 and 50 % Glycerol.

Konservierungsmittel: ProClin

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

Lagerung: -20 °C

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

Haltbarkeit: 12 months

Referenzen für anti-MYCN Antikörper (ABIN760676)

Ramraj, Aravindan, Somasundaram, Herman, Natarajan, Aravindan: "Serum-circulating miRNAs predict neuroblastoma progression in mouse model of high-risk metastatic disease." in: Oncotarget, Vol. 7, Issue 14, pp. 18605-19, (2016) (PubMed).

Details zu MYCN

Target: MYCN (N-Myc Proto-Oncogene Protein (MYCN))

Andere Bezeichnung: N-Myc

Hintergrund:

This gene is a member of the MYC family and encodes a protein with a basic helix-loop-helix (bHLH) domain. This protein is located in the nucleus and must dimerize with another bHLH protein in order to bind DNA. Amplification of this gene is associated with a variety of tumors, most notably neuroblastomas. [provided by RefSeq, Jul 2008].

Subcellular location: Nucleus

Synonyms: NMYC, ODED, MODED, N-myc, bHLHe37, N-myc proto-oncogene protein, Class E basic helix-loop-helix protein 37, MYCN

Gen-ID: 4613

UniProt: P04198