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Western blot
ImmunohistochemistryAntibody data
- Antibody Data
- Antigen structure
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- Validations
- Western blot [1]
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- Product number
- PA5-32527 - Provider product page
- Provider
- Invitrogen Antibodies
- Product name
- Anti-Osteopontin Polyclonal Antibody
- Antibody type
- Polyclonal
- Antigen
- Synthetic peptide
- Description
- Heat-mediated antigen retrieval is recommended prior to staining, using a 10mM citrate buffer, pH 6.0, for 10 minutes followed by cooling at room temperature for 20 min. Following antigen retrieval, incubate samples with primary antibody for 10 min at room temperature. A suggested positive control is breast carcinoma.
- Reactivity
- Human
- Host
- Rabbit
- Isotype
- IgG
- Vial size
- 500 µL
- Storage
- Store at 4°C short term. For long term storage, store at -20°C, avoiding freeze/thaw cycles.
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Supportive validation
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- Invitrogen Antibodies (provider)
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- Experimental details
- Western blot was performed using Anti-Osteopontin Polyclonal Antibody(Product # PA5-32527) and a 55kDa band corresponding to isoform of Osteopontin was observed in Hep G2 cell line treated with Protein Transport Inhibitor (PTI, 1X for 4 hr) as compared to IMR-32 cell line treated with same inhibitor. Whole cell extracts (30 µg lysate) of Hep G2 (Lane 1), Hep G2 + PTI (1X for 4 hr) (Lane 2), IMR-32 (Lane 3), IMR-32 + PTI (1X for 4 hr) (Lane 4) electrophoresed using NuPAGE™ 4-12% Bis-Tris Protein Gel (Product # NP0322BOX). Resolved proteins were then transferred onto a Nitrocellulose membrane (Product # IB23001) by iBlot® 2 Dry Blotting System (Product # IB21001). The blot was probed with the primary antibody (1:1000 dilution) and detected by chemiluminescence with Goat anti-Rabbit IgG (H+L) Superclonal™ Recombinant Secondary Antibody, HRP (Product # A27036,1:4000 dilution) using the iBright FL 1000 (Product # A32752). Chemiluminescent detection was performed using Novex® ECL Chemiluminescent Substrate Reagent Kit (Product # WP20005).
