IGF2R antibody

Catalog No. ABIN501206

The Mouse Monoclonal anti-IGF2R antibody has been validated for WB and FACS. It is suitable to detect IGF2R in samples from Human.

Quick Overview for IGF2R antibody (ABIN501206)

Target: IGF2R (Insulin-Like Growth Factor 2 Receptor (IGF2R))

Reactivity: Human

Host: Mouse

Clonality: Monoclonal

Conjugate: This IGF2R antibody is un-conjugated

Application: Western Blotting (WB), Flow Cytometry (FACS)

Clone: MEM-238

Product Details anti-IGF2R Antibody

Specificity: This antibody reacts with CD222 antigen (250 kDa) on Western blotting and Flow Cytometry.

Cross-Reactivity (Details): Species reactivity (tested):Human.

Purification: Protein-A Sepharose Chromatography of hybridoma supernatant

Immunogen: Recombinant vaccinia virus containing CD222 Remarks: Hybridoma was established by fusion of mouse myeloma cell Sp2/0-Ag14 with Balb/c mouse splenocyte

Isotype: IgG1

Application Details

Application Notes: Western blotting: 10 μg/mL for chemiluminescence detection system. Positive Controls: Jurkat, Raji cells. Flow Cytometry: 10 μg /mL (final concentration). Detailed procedure is provided in Protocols.
Other applications not tested.
Optimal dilutions are dependent on conditions and should be determined by the user.

Protocol: SDS-PAGE & Western Blotting1) Wash the cells 3 times with PBS and suspend with 10 volume of cold Lysis buffer (50 mMTris-HCl, pH 7. 2, 250 mM NaCl, 0. 1% NP-40, 2 mM EDTA, 10%glycerol) containingappropriate protease inhibitors. Incubate it at 4°C with rotating for 30 minutes, thensonicate briefly (up to 10 seconds). 2) Centrifuge the tube at 12,000 x g for 10 minutes at 4°C and transfer the supernatant toanother tube. Measure the protein concentration of the supernatant and add the cold Lysisbuffer to make 8 mg/mL solution. 3) Mix the sample with equal volume of Laemmli's sample buffer. 4) Boil the samples for 3 minutes and centrifuge. Load 10 μL of the sample per lane in a 1mm thick SDS-polyacrylamide gel for electrophoresis. 5) Blot the protein to a polyvinylidene difluoride (PVDF) membrane at 1 mA/cm2 for 1 hourin a semi-dry transfer system (Transfer Buffer: 25 mM Tris, 190 mM glycine, 20% MeOH). See the manufacture's manual for precise transfer procedure. 6) To reduce nonspecific binding, soak the membrane in 10% skimmed milk (in PBS, pH7. 2) for 1 hour at room temperature, or overnight at 4°C. 7) Incubate the membrane with primary antibody diluted with PBS, pH 7. 2 containing 1%skimmed milk as suggest in the APPLICATIONS for 1 hour at room temperature. (Theconcentration of antibody will depend on condition. )8) Wash the membrane with PBS-T [0. 05% Tween-20 in PBS] (5 minutes x 3 times). 9) Incubate the membrane with the 1: 10,000 HRP-conjugated anti-mouse IgG diluted with1% skimmed milk (in PBS, pH 7. 2) for 1 hour at RT. 10) Wash the membrane with PBS-T (10 minutes x 3 times). 11) Wipe excess buffer on the membrane, then incubate it with appropriatechemiluminescence reagent for 1 minute. 12) Remove extra reagent from the membrane by dabbing with paper towel, and seal it inplastic wrap. 13) Expose to an X-ray film in a dark room for 3 minutes. 14) Develop the film as usual. The condition for exposure and development may vary. Positive Controls: Jurkat, Raji. Flow cytometric analysis for floating cellsWe usually use Fisher tubes or equivalents as reaction tubes for all step described below. 1) Wash the cells 3 times with washing buffer [PBS containing 2% fetal calf serum (FCS) and0. 1% NaN3]2) Resuspend the cells with washing buffer (5x10e6 cells/mL). 3) Add 50 µL of the cell suspension into each tube, and centrifuge at 500 x g for 1 minute atRT (20~25°C). Remove supernatant by careful aspiration. 4) Add 10 µL of normal goat serum containing 1 mg/mL normal human IgG and 0. 1% NaN3to the cell pellet after tapping. Mix well and incubate at RT for 5 minutes. 5) Add 40 µL of the primary antibody at the concentration of as suggest in the

Restrictions: For Research Use only

Handling

Concentration: 1.0 mg/mL

Buffer: PBS, pH 7.2 containing 50 % Glycerol without preservatives.

Preservative: Without preservative

Handling Advice: Avoid repeated freezing and thawing.

Storage: -20 °C/-80 °C

Storage Comment: Store lyophilized (preferably in a desiccator) at -20 °C and in aliquots at -80 °C. Reconstituted antibody can be stored at 4 °C for a limited period of time, it does not show decline in activity after two weeks at 4 °C.

Target Details for IGF2R

Target: IGF2R (Insulin-Like Growth Factor 2 Receptor (IGF2R))

Alternative Name: CD222 / IGF2R

Background: The insulin-like growth factor 2 receptor (IGF-2R/CD222) is a single-chain protein of ~250 kDa. It is a high affinity binding protein for IGF-2 and shows very low affinity for IGF-1. This receptor is identical with the cation-independent mannose-6- phosphate receptor (MPR300) involved in lysosomal enzyme targeting, particularly in the transport of newly synthesized acid hydrolases to lysomes. Binding of mannose and IGF-2 are independent of each other and utilize two different binding sites.Synonyms: 300 kDa mannose 6-phosphate receptor, CI Man-6-P receptor, CI-MPR, Insulin-like growth factor 2 receptor, Insulin-like growth factor II receptor, Late Endosome Marker, M6P/IGF2 receptor, M6P/IGF2R, M6PR, MPR 300, Mannose 6 Phosphate Receptor (Cation independent)

Gene ID: 3482

NCBI Accession: NP_000867

UniProt: P11717