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Western blot
ImmunocytochemistryAntibody data
- Antibody Data
- Antigen structure
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- Validations
- Western blot [3]
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- Product number
- PA5-19689 - Provider product page
- Provider
- Invitrogen Antibodies
- Product name
- Anti-eIF3b Polyclonal Antibody
- Antibody type
- Polyclonal
- Antigen
- Synthetic peptide
- Description
- This antibody is predicted to react with mouse, rat, cow and fruit fly (Drosophila melanogaster) based on sequence homology.
- Reactivity
- Human, Mouse
- Host
- Rabbit
- Isotype
- IgG
- Vial size
- 100 µg
- Concentration
- 0.7 mg/mL
- 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
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Western blot analysis of HepG2 Whole Cell Lysate using Product # PA5-19689, eIF3B primary antibody at a dilution of 1 µg/mL. Blot treated with a secondary IR Dye680-conjugated Goat polyclonal anti-Rabbit antibody was used at a dilution of 1:10000.
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- KD of eIF3b was achieved by transfecting A-431 with eIF3b specific siRNAs (Silencer® select Product # s16492, s16493). Western blot analysis (Fig. a) was performed using whole cell extracts from the eIF3b KD cells (Lane 3), non-specific scrambled siRNA transfected cells (Lane 2) and untransfected cells (Lane 1). The blot was probed with eIF3b Polyclonal Antibody (Product # PA5-19689, 1 µg/mL) and Goat anti-Rabbit IgG (H+L) Superclonal™ Secondary Antibody, HRP conjugate (Product # A27036, 1:4000 dilution). Densitometric analysis of this western blot is shown in histogram (Fig. b). Decrease in signal upon siRNA mediated knock down confirms that antibody is specific to eIF3b. .
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Western blot was performed using Anti-eIF3b Polyclonal Antibody (Product # PA5-19689) and 110 kDa band corresponding to eIF3b was observed across cell lines tested. Whole cell extracts (30 µg lysate) of A-431 (Lane 1), Hep G2 (Lane 2), HEK-293 (Lane 3), NIH/3T3 (Lane 4), K-562 (Lane 5), Jurkat (Lane 6), DU 145 (Lane 7), PANC-1 (Lane 8) and SH-SY5Y (Lane 9) were 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 µg/mL) and detected by chemiluminescence Goat Anti-Rabbit IgG Secondary Antibody, HRP conjugate (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).
