GRB2 Antikörper

Produktnummer ABIN686932

Produktdetails anti-GRB2 Antikörper

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

Reaktivität: Human, Maus, Ratte, Hund, Huhn

Wirt: Kaninchen

Klonalität: Polyklonal

Konjugat: Dieser GRB2 Antikörper ist unkonjugiert

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

Kreuzreaktivität: Human, Ratte

Homologie: Mouse,Dog,Chicken

Aufreinigung: Purified by Protein A.

Immunogen: KLH conjugated synthetic peptide derived from human GRB2

Isotyp: IgG

Anwendungsinformationen

Applikationshinweise: WB(1:100-500)
Optimal working dilution should be determined by the investigator.

Beschränkungen: Nur für Forschungszwecke einsetzbar

Independent Validation (WB)

Validierung #029728 (Western Blotting)

by: Alamo Laboratories Inc

No.: #029728

Datum: 05.06.2014

Chargennummer: 121228

Validierte Anwendung: Western Blotting

Positivkontrolle: NIH/3T3 cells

Negativkontrolle: HeLa cells - very low expression

Bewertung: 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.

Validierungsbilder

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).

Protokoll

Primärantikörper:

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

Sekundärantikörper:

• 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).

Anmerkungen: Nothing to note.

Handhabung

Format: Liquid

Konzentration: 1 mg/mL

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

Konservierungsmittel: Sodium azide

Vorsichtsmaßnahmen: This product contains sodium azide: a POISONOUS AND HAZARDOUS SUBSTANCE which should be handled by trained staff only.

Lagerung: 4 °C,-20 °C

Informationen zur Lagerung: Shipped at 4°C. Store at -20°C for one year. Avoid repeated freeze/thaw cycles.

Haltbarkeit: 12 months

Details zu GRB2

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

Andere Bezeichnung: GRB2/ASH (GRB2 Produkte)

Synonyme: GRB2 antikoerper, grb2 antikoerper, AA408164 antikoerper, Ash antikoerper, ASH antikoerper, EGFRBP-GRB2 antikoerper, Grb3-3 antikoerper, MST084 antikoerper, MSTP084 antikoerper, NCKAP2 antikoerper, wu:fb18e12 antikoerper, wu:fb34e06 antikoerper, zgc:73077 antikoerper, growth factor receptor bound protein 2 antikoerper, growth factor receptor-bound protein 2 antikoerper, growth factor receptor-bound protein 2b antikoerper, GRB2 antikoerper, grb2 antikoerper, Grb2 antikoerper, grb2b antikoerper

Hintergrund: 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.

Molekulargewicht: 25kDa

Gen-ID: 2885

UniProt: P62993

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