MEK2 anticorps (AA 1-50)

N° du produit ABIN726500

L’anticorps Lapin Polyclonal anti-MEK2 a été validé pour WB, ELISA, IHC (p), FACS, IF (cc), IF (p) et IHC (fro) et a été validé indépendamment. Il convient pour détecter MEK2 dans des échantillons de Humain et Souris.

Aperçu rapide pour MEK2 anticorps (AA 1-50) (ABIN726500)

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

Reactivité: Humain, Souris

Hôte: Lapin

Clonalité: Polyclonal

Conjugué: Cet anticorp MEK2 est non-conjugé

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))

Détail du produit anti-MEK2 anticorps

Épitope: AA 1-50

Réactivité croisée: Humain, Souris

Homologie: Rat,Chicken

Purification: Purified by Protein A.

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

Isotype: IgG

Information d'application

Indications d'application: 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: 02.07.2014

Numéro du lot: 110511

Application validée: Western Blotting

Contrôle positif: Hela cell extract

Contrôle négative: c6/36 Mosquito cell extract (non-reactive species)

Conclusion: 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).

Protocole

Anticorps primaire:

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

Anticorps secondaire:

• 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).

Notes: - No challenges noted.

Stockage

Format: Liquid

Concentration: 1 μg/μL

Buffer: 0.01M TBS( pH 7.4) with 1 % BSA, 0.02 % Proclin300 and 50 % Glycerol.

Agent conservateur: ProClin

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

Stock: 4 °C,-20 °C

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

Date de péremption: 12 months

Détails sur MEK2

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

Autre désignation: MEK2

Sujet:

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).

ID gène: 5605

UniProt: P36507

Pathways: Signalisation MAPK, Signalisation RTK, 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