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Western blot analysis of Nucleoporin p62 was performed by loading whole cell lysates (20ug) of the indicated samples and 5ul of Thermo Scientific PageRuler Plus Prestained Protein Ladder (Product # 26619) onto a 4-20% Tris-Glycine polyacrylamide gel. Proteins were transferred to a nitrocellulose membrane using the G2 Fast Blotter (Product # 62288), and blocked with 5% milk in TBST for 1 hour at room temperature. Nucleoporin p62 was detected at ~62kD in samples using a Nucleoporin p62 (RL31) monoclonal antibody (Product # MA1-073) at a dilution of 1:200 overnight at 4C on a rocking platform, followed by a goat anti-armenian hamster IgG-HRP secondary antibody (Product # PA1-32045) at a dilution of 1:10,000 for at least 30 minutes at room temperature. Chemiluminescent detection was performed using SuperSignal West Pico substrate (Product # 34080).
|Tested species reactivity||Dog, Human, Mouse, Primate, Rat|
|Host / Isotype||Armenian hamster / IgG|
|Immunogen||O-linked glycoprotein-enriched fraction from rat liver nuclear envelopes raised in Armenian hamsters.|
|Storage buffer||PBS, pH 7.2-7.4, with 30% glycerol, 1mg/ml BSA|
|Contains||0.05% sodium azide|
|Storage Conditions||-20° C, Avoid Freeze/Thaw Cycles|
|Tested Applications||Dilution *|
|Western Blot (WB)||1:200|
* Suggested working dilutions are given as a guide only. It is recommended that the user titrate the product for use in their own experiment using appropriate negative and positive controls.
MA1-073 detects the nucleoporin p62 O-linked glycoprotein in the p62 complex from rat, human and non-human primate tissues.
MA1-073 has been successfully used in Western blot and immunofluorescence procedures. By Western blot, this antibody detects a single ~62 kDa protein representing nucleoporin p62 in rat liver nuclear envelope fractions. Immunofluorescence staining of p62 in rat liver with MA1-073 results in staining of both the nucleoplasmic and cytoplasmic surfaces of the nuclear pore complex (NPC).
The MA1-073 immunogen is O-linked glycoprotein-enriched fraction from rat liver nuclear envelopes raised in Armenian hamsters.
Diffusion of metabolites and small non-nuclear molecules as well as active, mediated import of protein and export of protein and RNA through the nuclear envelope occurs through nuclear pore complexes or NPC"e;s. NPC"e;s contain up to 100 different polypeptides which have a combined mass of about 125 megadaltons. The channel available for passive transport through the NPC is about 9-10 nm in diameter while carrier mediated changes in the NPC result in a ~25 nm channel used for larger, actively transported molecules. Of the 100 polypeptides, at least 8 of these are O-linked N-acetylglycosamine-modified in mammalian cells. All of the mammalian O-linked glycoproteins contain multiple copies of phenylalanine, glycine dipeptide repeats dispersed throughout part of their sequence. Studies indicate that the NPC O-linked glycoproteins have a direct role in nuclear protein import.
For Research Use Only. Not for use in diagnostic procedures. Not for resale without express authorization.
62 kDa nucleoporin; nuclear pore complex 1; nuclear pore glycoprotein 62; nuclear pore glycoprotein p62; nucleoporin 62kD; nucleoporin 62kDa; nucleoporin Nup62; NUP62
AA589433; AI426861; AI790512; AU045898; D7Ertd649e; IBSN; Np62; NUP62; Nupc1; p62; SNDI