MEK2 antibody (AA 1-50)

Catalog No. ABIN726500

The Rabbit Polyclonal anti-MEK2 antibody has been validated for WB, ELISA, IHC (p), FACS, IF (cc), IF (p) and IHC (fro) and has been independently validated. It is suitable to detect MEK2 in samples from Human and Mouse.

Quick Overview for MEK2 antibody (AA 1-50) (ABIN726500)

Target: MEK2 (MAP2K2) (Mitogen-Activated Protein Kinase Kinase 2 (MAP2K2))

Reactivity: Human, Mouse

Host: Rabbit

Clonality: Polyclonal

Conjugate: This MEK2 antibody is un-conjugated

Application: Western Blotting (WB), ELISA, Immunohistochemistry (Paraffin-embedded Sections) (IHC (p)), Flow Cytometry (FACS), Immunofluorescence (Cultured Cells) (IF (cc)), Immunofluorescence (Paraffin-embedded Sections) (IF (p)), Immunohistochemistry (Frozen Sections) (IHC (fro))

Product Details anti-MEK2 Antibody

Binding Specificity: AA 1-50

Cross-Reactivity: Human, Mouse

Predicted Reactivity: Rat,Chicken

Purification: Purified by Protein A.

Immunogen: KLH conjugated synthetic peptide derived from human MAPKK2/MEK2

Isotype: IgG

Application Details

Application Notes: WB 1:300-5000
ELISA 1:500-1000
FCM 1:20-100
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 #029748 (Western Blotting)

by: Alamo Laboratories Inc

No.: #029748

Date: 07/02/2014

Lot Number: 110511

Method validated: Western Blotting

Positive Control: Hela cell extract

Negative Control: c6/36 Mosquito cell extract (non-reactive species)

Notes: A single positive band at the correct molecular weight was detected in positive control HeLa cell extract. Several bands were observed in the non-reactive species negative control at approximately the same molecular weight as MAP2K2. These bands may constitute cross-reactivity of the target antigen, or they may be non-specific bands.

Validation Images

Figure 1. Western blot of lysates from HeLa cells (Lane 1) and c6/36 cells (Lane 2) probed with anti-MAP2K2 (upper panel) or with anti-Actin for loading control (lower panel).

Full Methods

Primary Antibody:

• Antigen: Mitogen-Activated Protein Kinase Kinase 2 (MAP2K2) (1:150 dilution)
• Catalog number: ABIN726500
• Lot number: 110511

Secondary Antibody:

• Antibody: Goat Anti-Rabbit IgG (H + L)-HRP Conjugate (1:20,000 dilution)
• Lot number: L170-6515

Full Protocol:

• 1. Total protein extracts were boiled in 1X SDS Sample Buffer containing 1% SDS and 1.25% β-mercaptoethanol at 95°C for 5 min prior to loading.
• 2. 46 μg of boiled extracts 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) non-fat dry milk at room temperature for 1 h.
• 6. The membrane was rinsed with TBST once.
• 7. The membrane was immersed with the protein side up in the primary antibody solution (anti-MAP2K2; 1:150) in TBST containing 5% (W/V) non-fat dry milk and incubated for 16 h at 4°C.
• 8. The membrane was rinsed in TBST thrice for 5 min each.
• 9. The membrane was incubated in the HRP-conjugated secondary antibody solution (Goat anti-rabbit IgG-HRP; 1:20,000) in TBST containing 5% (W/V) non-fat dry milk and incubated for 1 h at room temperature (~26°C) with gentle agitation.
• 10. The membrane was rinsed thrice TBST thrice for 5 min each.
• 11. The membrane was rinsed in TBS twice for 30 s each.
• 12. Signals were detected with ECL-2 Substrate. The blot was scanned for 300 s.
• 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 min each.
• 15. The membrane was washed in TBST 2 times for 10 min each.
• 16. Repeated Steps 5-12 with the loading control antibody (anti-Actin; 1:6,000) and its matching secondary antibody (Goat anti-rabbit IgG-HRP; 1:20,000).

Experimental Notes: - No challenges noted.

Handling

Format: Liquid

Concentration: 1 μg/μL

Buffer: 0.01M TBS( pH 7.4) with 1 % BSA, 0.02 % 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: 4 °C,-20 °C

Storage Comment: Shipped at 4°C. Store at -20°C for one year. Avoid repeated freeze/thaw cycles.

Expiry Date: 12 months

Target Details for MEK2

Target: MEK2 (MAP2K2) (Mitogen-Activated Protein Kinase Kinase 2 (MAP2K2))

Alternative Name: MEK2

Background:

Synonyms: CFC4, MEK2, MKK2, MAPKK2, PRKMK2, Dual specificity mitogen-activated protein kinase kinase 2, MAP kinase kinase 2, MAPKK 2, ERK activator kinase 2, MAPK/ERK kinase 2, MEK 2, MAP2K2

Background: Catalyzes the concomitant phosphorylation of a threonine and a tyrosine residue in a Thr-Glu-Tyr sequence located in MAP kinases. Activates the ERK1 and ERK2 MAP kinases (By similarity).

Gene ID: 5605

UniProt: P36507

Pathways: MAPK Signaling, RTK Signaling, Fc-epsilon Receptor Signaling Pathway, Neurotrophin Signaling Pathway, Activation of Innate immune Response, Toll-Like Receptors Cascades, Signaling of Hepatocyte Growth Factor Receptor, BCR Signaling