GRB2 anticorps

N° du produit ABIN686932

Cet validé indépendamment anticorps Lapin Polyclonal détecte spécifiquement GRB2 dans ELISA et IHC (p). Il présente une réactivité envers Humain, Souris, Rat, Chien et Poulet.

Aperçu rapide pour GRB2 anticorps (ABIN686932)

Antigène: GRB2 (Growth Factor Receptor-Bound Protein 2 (GRB2))

Reactivité: Humain, Souris, Rat, Chien, Poulet

Hôte: Lapin

Clonalité: Polyclonal

Conjugué: Cet anticorp GRB2 est non-conjugé

Application: ELISA, Immunohistochemistry (Paraffin-embedded Sections) (IHC (p))

Détail du produit anti-GRB2 anticorps

Réactivité croisée: Humain, Rat

Homologie: Mouse,Dog,Chicken

Purification: Purified by Protein A.

Immunogène: KLH conjugated synthetic peptide derived from human GRB2

Isotype: IgG

Information d'application

Indications d'application: WB(1:100-500)
Optimal working dilution should be determined by the investigator.

Restrictions: For Research Use only

Independent Validation (WB)

Validation #029728 (Western Blotting)

by: Alamo Laboratories Inc

No.: #029728

Date: 05.06.2014

Numéro du lot: 121228

Application validée: Western Blotting

Contrôle positif: NIH/3T3 cells

Contrôle négative: HeLa cells - very low expression

Conclusion: A strong band was seen at the correct molecular weight in the positive control, and no band was present in the negative control. Additionally, no non-specific bands were observed.

Validation Images

Figure 1: Western blot of lysates from NIH/3T3 cells (Lane 1, positive control) and HeLa cells (Lane 2, negative control) probed with anti-GRB2 (upper panel) or with anti- Actin for loading control (lower panel).

Protocole

Anticorps primaire:

• Antigen: Growth Factor Receptor-Bound Protein 2 (GRB2) (1:200 dilution)
• Catalog number: ABIN686932
• Lot number: 121228

Anticorps secondaire:

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

Full Protocol:

• 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.
• 46 μg of boiled extracts were loaded and resolved on a 8-16% SDS-polyacrylamide gel.
• The Spectra Multicolor Broad Range (Thermo Scientific, Cat # 26634) were used as molecular mass markers.
• Proteins were transferred onto PVDF membrane by wet transfer and protein transfer was confirmed with Ponceau-S staining.
• 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.
• The membrane was rinsed with TBST once.
• The membrane was immersed with the protein side up in the primary antibody solution (anti-GRB2; 1:200) in TBST containing 5% (W/V) non-fat dry milk and incubated for 16 hours at 4°C.
• The membrane was rinsed in TBST three times for 5 min each.
• 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 hour at room temperature (~26°C) with gentle agitation.
• The membrane was rinsed three times with TBST for 5 min each.
• The membrane was rinsed in TBS twice for 30 s each.
• Signals were detected with ECL-2 Substrate. The blot was scanned for 300 s.
• The membrane was rinsed three times with TBST, then incubated in Acidic Glycine Stripping Buffer at room temperature with gentle agitation for 3 times, 10 min each.
• The membrane was washed in TBST 2 times for 10 min each.
• 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: Nothing to note.

Stockage

Format: Liquid

Concentration: 1 mg/mL

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

Agent conservateur: Sodium azide

Précaution d'utilisation: This product contains sodium azide: 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 GRB2

Antigène: GRB2 (Growth Factor Receptor-Bound Protein 2 (GRB2))

Autre désignation: GRB2/ASH

Sujet: Optional[synonyms]: GRB 2, GRB2 adapter protein, Grb3 3, Growth Factor Receptor Bound Protein 2, Growth factor receptor bound protein 3, EGFRBP GRB2, Growth factor receptor-bound protein 2, Abundant SRC homology, Adapter protein GRB2, ASH, Ash protein, EGFRBP GRB2, Epidermal growth factor receptor binding protein, Epidermal growth factor receptor binding protein GRB2, GRB 2, GRB2 adapter protein, Grb3 3, Growth Factor Receptor Bound Protein 2, Growth factor receptor bound protein 3, HT027, MST084, MSTP084, OTTHUMP00000166096, OTTHUMP00000166097, OTTHUMP00000166098, Protein ASH, SEM5, SH2/SH3 adapter GRB2.

Poids moléculaire: 25kDa

ID gène: 2885

UniProt: P62993

Pathways: Signalisation RTK, TCR Signaling, Fc-epsilon Receptor Signaling Pathway, EGFR Signaling Pathway, Neurotrophin Signaling Pathway, Regulation of Actin Filament Polymerization, Hepatitis C, Signaling Events mediated by VEGFR1 and VEGFR2, Signaling of Hepatocyte Growth Factor Receptor, EGFR Downregulation