MEK2 antibody (AA 1-50)

Catalog No. ABIN726500

Product Details anti-MEK2 Antibody

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

Binding Specificity: AA 1-50

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))

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 (MAP2K2 Products)

Synonyms: CFC4 antibody, MAPKK2 antibody, MEK2 antibody, MKK2 antibody, PRKMK2 antibody, AA589381 antibody, MK2 antibody, Prkmk2 antibody, ATMKK2 antibody, F27B13.50 antibody, F27B13_50 antibody, MAP KINASE KINASE 1 antibody, MAP kinase kinase 2 antibody, MK1 antibody, map2k2 antibody, wu:fa56c06 antibody, zgc:123171 antibody, zgc:172250 antibody, mitogen-activated protein kinase kinase 2 antibody, mitogen activated protein kinase kinase 2 antibody, MAP kinase kinase 2 antibody, mitogen-activated protein kinase kinase 2a antibody, mitogen-activated protein kinase kinase 2b antibody, MAP2K2 antibody, Map2k2 antibody, MKK2 antibody, map2k2a antibody, map2k2b antibody

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