PA1-084

antibody from Invitrogen Antibodies

Targeting: RANBP3

Western blot (Data presented on provider website)

Immunocytochemistry (Supportive data in Antibodypedia)

Immunohistochemistry (Recommended by provider)

Other assay (Supportive data in Antibodypedia)

Antibody Data

Product number

PA1-084

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. PA1-084 has been successfully used in Western blot and immunofluorescence procedures. By Western blot this antibody detects an ~60 kDa protein representing RanBP3 from HeLa cell extracts. Immunofluorescence staining of RanBP3 in NRK cells with PA1-084 results in diffuse nuclear staining. The PA1-084 immunogen is a synthetic peptide corresponding to residues M(1) A D L A N E E K P A L A(13) C of human RanBP3. The PA1-084 immunizing peptide (Cat. # PEP-177) is available for use in neutralization and control experiments.

Reactivity

Human, Rat

Host

Rabbit

Isotype

IgG

Vial size

200 µg

Concentration

1 mg/mL

Storage

-20° C, Avoid Freeze/Thaw Cycles

References

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

Immunocytochemistry

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Immunocytochemical staining of RanBP3 in NRK cells using Product # PA1-084.

Other assay

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.