IGF1R antibody (AA 251-350)

Catalog No. ABIN726575

Product Details anti-IGF1R Antibody

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

Binding Specificity: AA 251-350

Reactivity: Human, Mouse, Rat, Dog

Host: Rabbit

Clonality: Polyclonal

Conjugate: This IGF1R antibody is un-conjugated

Application: Flow Cytometry (FACS)

Cross-Reactivity: Dog, Human, Mouse, Rat

Predicted Reactivity: Cow,Sheep,Pig,Horse,Rabbit

Purification: Purified by Protein A.

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

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
ICC 1:100-500

Restrictions: For Research Use only

Independent Validation (WB)

Validation #029750 (Western Blotting)

by: Alamo Laboratories Inc

No.: #029750

Date: 07/01/2014

Lot Number: 130322

Method validated: Western Blotting

Positive Control: MCF-7 cells – high expression

Negative Control: PC3 cells – low expression

Notes: 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.

Validation Images

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).

Full Methods

Primary Antibody:

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

Secondary Antibody:

• 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).

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

References for anti-IGF1R antibody (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).

Target Details for IGF1R

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

Alternative Name: IGF1R (IGF1R Products)

Synonyms: CMT2N antibody, CD221 antibody, IGFIR antibody, IGFR antibody, JTK13 antibody, IGFIRC antibody, IGFR1 antibody, IGF-1R antibody, IGF1R antibody, DKFZp469I0618 antibody, igf-1r antibody, xIGF-1R antibody, xIGFR antibody, xigf1r antibody, A330103N21Rik antibody, D930020L01 antibody, hyft antibody, igf-ir antibody, IGF-IR antibody, IGF-IRb antibody, wu:fb03h06 antibody, IGF-IRa antibody, sb:eu1090 antibody, wu:fi20b10 antibody, alanyl-tRNA synthetase antibody, insulin like growth factor 1 receptor antibody, insulin-like growth factor 1 receptor antibody, insulin-like growth factor 1 antibody, insulin-like growth factor I receptor antibody, insulin like growth factor 1 receptor S homeolog antibody, insulin-like growth factor 1b receptor antibody, insulin-like growth factor 1a receptor antibody, AARS antibody, IGF1R antibody, Igf1r antibody, igf1r antibody, Igf1 antibody, LOC780503 antibody, LOC100136427 antibody, igf1r.S antibody, igf1rb antibody, igf1ra antibody

Background:

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.

Gene ID: 3480

UniProt: P08069

Pathways: RTK Signaling, Regulation of Hormone Metabolic Process, Regulation of Hormone Biosynthetic Process, Autophagy