Antibody data
- Antibody Data
- Antigen structure
- References [0]
- Comments [0]
- Validations
- Western blot [1]
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Validation data
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- Product number
- PA5-32580 - Provider product page
- Provider
- Invitrogen Antibodies
- Product name
- Anti-Phospho-S6 (Ser235, Ser236) 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 30 min at room temperature. A suggested positive control is breast carcinoma.
- Reactivity
- Human
- Host
- Rabbit
- Isotype
- IgG
- Vial size
- 500 µL
- Storage
- -20° C, Avoid 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-Phospho-S6 (Ser235, Ser236) Polyclonal Antibody(Product # PA5-32580) and a 32 kDa band corresponding to Phospho-S6 (Ser235, Ser236) was observed across the cell lines tested. Whole cell extracts (30 µg lysate) of U-2 OS (Lane 1), U-2 OS treated with IGF (Lane 2), U-2 OS treated with Rapamycin (Lane 3), U-2 OS treated with both IGF and Rapamycin (Lane 4), HEK-293 (Lane 5) were electrophoresed using NuPAGE™ 4-12% Bis-Tris Protein Gel (Product # NP0321BOX). Resolved proteins were then transferred onto a Nitrocellulose membrane (Product # LC2001) 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). IGF was used at a concentration of 50 ng/mL and Rapamycin at 50 nM strength.