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Western blotAntibody data
- Antibody Data
- Antigen structure
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- Validations
- Western blot [1]
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- Product number
- PA5-17782 - Provider product page
- Provider
- Invitrogen Antibodies
- Product name
- Phospho-NFkB p65 (Ser536) Polyclonal Antibody
- Antibody type
- Polyclonal
- Antigen
- Synthetic peptide
- Description
- It is not recommended to aliquot this antibody. This antibody is not cross-reactive with the p50 subunit or other related proteins.
- Reactivity
- Human, Mouse, Rat
- Host
- Rabbit
- Isotype
- IgG
- Vial size
- 100 µL
- Concentration
- 88 µg/mL
- Storage
- -20°C
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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-NFkB p65 (Ser536) Polyclonal Antibody (Product # PA5-17782) and a 65 kDa band corresponding to Transcription factor p65 was observed across the cell lines tested. Whole cell extracts (30 µg lysate) of HeLa (Lane 1), HeLa treated with TNF-alpha (20 ng/mL for 10 mins) and Calyculin A (100 nM for 10 min) (Lane 2), HeLa treated with LPS (1 µg/mL for 15 mins) and Calyculin A (100 nM for 10 min) (Lane 3), Jurkat (Lane 4), Jurkat treated with TNF-alpha (20 ng/mL for 10 mins) and Calyculin A (100 nM for 10 min) (Lane 5), THP-1 (Lane 6), THP-1 treated with LPS (1 µg/mL for 15 mins) and Calyculin A (100 nM for 10 min) (Lane 7), RAW 264.7 (Lane 8), RAW 264.7 treated with LPS (1 µg/mL for 15 mins) and Calyculin A (100 nM for 10 min) (Lane 9) were electrophoresed using NuPAGE™ 4-12% Bis-Tris Protein Gel (Product # NP0321BOX). 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, 1:10,000 dilution) using the iBright FL 1000 (Product # A32752). Chemiluminescent detection was performed using Novex® ECL Reagent Kit (Product # WP20005).
