MYCN antibody (AA 401-464)

Catalog No. ABIN760676

Product Details anti-MYCN Antibody

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

Binding Specificity: AA 401-464

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))

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 (MYCN Products)

Synonyms: MODED antibody, MYCNOT antibody, N-myc antibody, NMYC antibody, ODED antibody, bHLHe37 antibody, Nmyc antibody, Nmyc-1 antibody, Nmyc1 antibody, c-nmyc antibody, Nmuc1 antibody, fb57a02 antibody, nmyc1 antibody, wu:fb57a02 antibody, zgc:85706 antibody, XN-myc antibody, XNmyc antibody, mycn antibody, mycn-a antibody, mycn-b antibody, mycna antibody, xN-myc1 antibody, MYCN proto-oncogene, bHLH transcription factor antibody, v-myc avian myelocytomatosis viral oncogene neuroblastoma derived homolog antibody, v-myc avian myelocytomatosis viral related oncogene, neuroblastoma derived antibody, olfactory receptor 8S1 antibody, MYCN proto-oncogene, bHLH transcription factor S homeolog antibody, MYCN antibody, Mycn antibody, LOC515887 antibody, mycn antibody, mycn.S antibody

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