MYCN antibody (AA 401-464)

Catalog No. ABIN760676

The Rabbit Polyclonal anti-MYCN antibody has been validated for ELISA, IF (cc), IF (p), IHC (p) and IHC (fro) and has been independently validated. It is suitable to detect MYCN in samples from Human and Mouse. There is 1 publication available.

Quick Overview for MYCN antibody (AA 401-464) (ABIN760676)

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

Reactivity: Human, Mouse

Host: Rabbit

Clonality: Polyclonal

Conjugate: This MYCN antibody is un-conjugated

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

Product Details anti-MYCN Antibody

Binding Specificity: AA 401-464

Predicted Reactivity: Rat,Cow,Pig

Purification: Purified by Protein A.

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

Isotype: IgG

Application Details

Application Notes: 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

Restrictions: For Research Use only

Independent Validation (WB)

Validation #029800 (Western Blotting)

by: Alamo Laboratories Inc

No.: #029800

Date: 08/21/2014

Lot Number: 120917

Method validated: Western Blotting

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

Negative Control: SK-N-SH cells - RNA

Notes: 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.

Validation Images

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

Full Methods

Primary Antibody:

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

Secondary Antibody:

• 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.

Experimental Notes: - No experimental challenges noted.

Handling

Format: Liquid

Concentration: 1 μg/μL

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

Preservative: ProClin

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

Storage: -20 °C

Storage Comment: Store at -20°C for one year. Avoid repeated freeze/thaw cycles.

Expiry Date: 12 months

References for anti-MYCN antibody (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).

Target Details for MYCN

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

Alternative Name: N-Myc

Background:

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

Gene ID: 4613

UniProt: P04198