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Western blotAntibody data
- Antibody Data
- Antigen structure
- References [1]
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- Validations
- Western blot [1]
- Immunocytochemistry [1]
- Other assay [1]
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- Product number
- PA1-084 - Provider product page
- Provider
- Invitrogen Antibodies
- Product name
- RANBP3 Polyclonal Antibody
- Antibody type
- Polyclonal
- Antigen
- Synthetic peptide
- Description
- PA1-084 detects Ran binding protein 3 (RanBP3) from human and rat cells.
- Concentration
- 1 mg/mL
Submitted references Sphingosine kinase 1 serves as a pro-viral factor by regulating viral RNA synthesis and nuclear export of viral ribonucleoprotein complex upon influenza virus infection.
Seo YJ, Pritzl CJ, Vijayan M, Bomb K, McClain ME, Alexander S, Hahm B
PloS one 2013;8(8):e75005
PloS one 2013;8(8):e75005
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Supportive validation
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- Invitrogen Antibodies (provider)
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- Experimental details
- Western blot analysis was performed on nuclear enriched extracts of A549 (Lane 1), THP-1 (Lane 2), Jurkat (Lane 3), Ramos (Lane 4), HEK-293 (Lane 5) and tissue extract of Rat Testis (Lane 6). The blot was probed with Anti-RANBP3 Rabbit Polyclonal Antibody (Product # PA1-084, 2µg/mL) and detected by chemiluminescence using Goat anti-Rabbit IgG (H+L) Superclonal™ Secondary Antibody, HRP conjugate (Product # A27036, 0.25µg/mL, 1:4000 dilution). A 60 kDa band corresponding to RANBP3 was observed across cell lines and tissue tested. Known quantity of protein samples were electrophoresed using Novex® NuPAGE® 4-12 % Bis-Tris gel (Product # NP0321BOX), XCell SureLock™ Electrophoresis System (Product # EI0002) and Novex® Sharp Pre-Stained Protein Standard (Product # LC5800). Resolved proteins were then transferred onto a nitrocellulose membrane with iBlot® 2 Dry Blotting System (Product # IB21001). The membrane was probed with the relevant primary and secondary Antibody following blocking with 5 % skimmed milk. Chemiluminescent detection was performed using Pierce™ ECL Western Blotting Substrate (Product # 32106).
Supportive validation
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Immunocytochemical staining of RanBP3 in NRK cells using Product # PA1-084.
Supportive validation
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Figure 5 SK inhibition impairs virus-induced activation of ERK/p90RSK/AKT to inhibit RanBP3-mediated nuclear export of viral NP. (A and B) HEK293 cells were transfected with scramble siRNA (siCTR) or siRNA targeting RanBP3 (siRanBP3); then the cells were infected with influenza virus at an MOI of 0.01 (A) or 1 (B). RanBP3, pRanBP3, viral M1, and alpha-tubulin were detected by Western blotting at 30 hpi (A). Viral NP (green) and nuclei (DRAQ5 dye, red) were visualized by confocal microscopy at 12 hpi (B). (C and D) MDCK cells were treated with solvent, DMS (5 uM), or SKI-II (10 uM) and infected with influenza virus at an MOI of 5 for the indicated times (C) or 6 hrs (D). Cell lysates were used for Western blot analysis to detect pRanBP3, RanBP3, alpha-tubulin, or actin. (E) MDCK cells were left untreated or treated with U0126 (10 uM) or LY294002 (10 uM) and infected with influenza virus at 3 MOI for 6 hrs. Western blot analysis was performed to detect pRanBP3, RanBP3, and GAPDH. (F) MDCK cells were treated with solvent alone or SKI-II (10 uM) and infected with influenza virus at an MOI of 1. Western blot analysis was performed to detect pERK, ERK, p-p90RSK, p90RSK, pRANBP3, or alpha-tubulin at 12 hpi. (G) MDCK cells were treated with solvent or SKI-II (10 uM) and mock-infected or infected with influenza virus at an MOI of 3. At 8 hpi, pAKT, AKT, and alpha-tubulin were detected by Western blot analysis.
