IGF1R Antikörper (AA 251-350)

Produktnummer ABIN726575

Der Kaninchen Polyklonal Anti-IGF1R-Antikörper wurde für FACS validiert und wurde unabhängig validiert. Er ist geeignet, IGF1R in Proben von Human, Maus, Ratte und Hund zu detektieren. Es sind 2+ Publikationen verfügbar.

Kurzübersicht für IGF1R Antikörper (AA 251-350) (ABIN726575)

Target: IGF1R (Insulin-Like Growth Factor 1 Receptor (IGF1R))

Reaktivität: Human, Maus, Ratte, Hund

Wirt: Kaninchen

Klonalität: Polyklonal

Konjugat: Dieser IGF1R Antikörper ist unkonjugiert

Applikation: Flow Cytometry (FACS)

Produktdetails anti-IGF1R Antikörper

Bindungsspezifität: AA 251-350

Kreuzreaktivität: Hund, Human, Maus, Ratte

Homologie: Cow,Sheep,Pig,Horse,Rabbit

Aufreinigung: Purified by Protein A.

Immunogen: KLH conjugated synthetic peptide derived from human IGF-1R alpha chain

Isotyp: IgG

Anwendungsinformationen

Applikationshinweise: 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
ICC 1:100-500

Beschränkungen: Nur für Forschungszwecke einsetzbar

Independent Validation (WB)

Validierung #029750 (Western Blotting)

by: Alamo Laboratories Inc

No.: #029750

Datum: 01.07.2014

Chargennummer: 130322

Validierte Anwendung: Western Blotting

Positivkontrolle: MCF-7 cells – high expression

Negativkontrolle: PC3 cells – low expression

Bewertung: Two strong bands were observed in the positive control, one at the expected molecular weight, and one slightly higher (which may represent a glycosylated or precursor form). Weaker bands of the same molecular weight appear in the negative control, which is consistent with the expected lower expression in PC3 cells. No other major bands are present in either positive or negative controls.

Validierungsbilder

Figure 1. Western blot of lysates from MCF7 cells (Lane 1) and PC-3 cells (Lane 2) probed with anti-IGF1R (upper panel) or with anti-Actin for loading control (lower panel).

Protokoll

Primärantikörper:

• Antigen: Insulin-Like Growth Factor 1 Receptor (IGF1R) (1:200 dilution)
• Catalog number: ABIN726575
• Lot number: 130322

Sekundärantikörper:

• Antibody: 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-IGF1R; 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: - No challenges noted.

Handhabung

Format: Liquid

Konzentration: 1 μg/μL

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

Konservierungsmittel: ProClin

Vorsichtsmaßnahmen: This product contains ProClin: 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

Referenzen für anti-IGF1R Antikörper (ABIN726575)

Hu, Li, Hu, Wu, Li, Meng: "MicroRNA let-7a and let-7f as novel regulatory factors of the sika deer (Cervus nippon) IGF-1R gene." in: Growth factors (Chur, Switzerland), Vol. 32, Issue 1, pp. 27-33, (2014) (PubMed).

Yang, Liu, Li, Wen, Zhu, Xu: "Insulin-Like Growth Factor-1 Modulates Polycomb Cbx8 Expression and Inhibits Colon Cancer Cell Apoptosis." in: Cell biochemistry and biophysics, (2014) (PubMed).

Details zu IGF1R

Target: IGF1R (Insulin-Like Growth Factor 1 Receptor (IGF1R))

Andere Bezeichnung: IGF1R

Hintergrund:

Synonyms: IGFR, CD221, IGFIR, JTK13, Insulin-like growth factor 1 receptor, Insulin-like growth factor I receptor, IGF-I receptor, IGF1R

Background: When present in a hybrid receptor with INSR, binds IGF1. PubMed:12138094 shows that hybrid receptors composed of IGF1R and INSR isoform Long are activated with a high affinity by IGF1, with low affinity by IGF2 and not significantly activated by insulin, and that hybrid receptors composed of IGF1R and INSR isoform Short are activated by IGF1, IGF2 and insulin. In contrast, PubMed:16831875 shows that hybrid receptors composed of IGF1R and INSR isoform Long and hybrid receptors composed of IGF1R and INSR isoform Short have similar binding characteristics, both bind IGF1 and have a low affinity for insulin.

Gen-ID: 3480

UniProt: P08069

Pathways: RTK Signalweg, Regulation of Hormone Metabolic Process, Regulation of Hormone Biosynthetic Process, Autophagie