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Transportin (N-Term) antibody

Details for Product No. ABIN967620
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Antigen
Synonyms CG7398, Dmel\\CG7398, IMPbeta2, TRN, Tm, Trn1, dTRN
Epitope
N-Term
Reactivity
Human
Host
Mouse
Clonality (Clone)
Monoclonal ()
Application
Western Blotting (WB), BioImaging (BI)
Pubmed 4 references available
Quantity 0.1 mg
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Catalog No. ABIN967620
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Immunogen Human Transportin
Clone L22FA2
Isotype IgG1, kappa
Cross-Reactivity Rat (Rattus)
Characteristics 1. Please refer to us for technical protocols.
2. Since applications vary, each investigator should titrate the reagent to obtain optimal results.
3. This antibody has been developed and certified for the bioimaging application. However, a routine bioimaging test is not performed on every lot. Researchers are encouraged to titrate the reagent for optimal performance.
4. Caution: Sodium azide yields highly toxic hydrazoic acid under acidic conditions. Dilute azide compounds in running water before discarding to avoid accumulation of potentially explosive deposits in plumbing.
5. Triton is a trademark of the Dow Chemical Company.
Purification Purified from tissue culture supernatant or ascites by affinity chromatography.
Background Transport of macromolecules into and out of the nucleus occurs via interaction of several cytosolic and nuclear pore proteins. The nuclear pore complex (NPC), anchored in the nuclear envelope, mediates active transport of proteins and RNA into and out of the nucleus. Transportin is 890 amino acids in length and a member of the karyopherin b (also known as importin b) superfamily of soluble transport proteins. It specifically interacts with a ~38-amino acid basic domain (M9) of its cargoes (proteins and ribonucleoproteins), the polypeptides of the NPC, and RanGTP (the GTP-bound form of the Ras family protein Ran), which modulates cargo binding. The gradient of RanGTP across the nuclear envelope regulates the transport process.
Synonyms: karyopherin beta2, importin beta2, M9 region interaction protein
Molecular Weight 95 - 101 kDa
Research Area Transporters
Application Notes Bioimaging
1. Seed the cells in appropriate culture medium at ~10,000 cells per well in an 96-well Imaging Plate and culture overnight.
2. Remove the culture medium from the wells, and fix the cells by adding 100 myl of Fixation Buffer to each well. Incubate for 10 minutes at room temperature (RT).
3. Remove the fixative from the wells, and permeabilize the cells using either 90% methanol, or Triton™ X-100: a. Add 100 myl of -20°C 90% methanol to each well and incubate for 5 minutes at RT. OR b. Add 100 myl of 0.1% Triton™ X-100 to each well and incubate for 5 minutes at RT.
4. Remove the permeabilization buffer, and wash the wells twice with 100 myl of 1× PBS.
5. Remove the PBS, and block the cells by adding 100 myl of to each well. Incubate for 30 minutes at RT.
6. Remove the blocking buffer and add 50 myl of the optimally titrated primary antibody (diluted in Stain Buffer) to each well, and incubate for 1 hour at RT.
7. Remove the primary antibody, and wash the wells three times with 100 myl of 1× PBS.
8. Remove the PBS, and add the second step reagent at its optimally titrated concentration in 50 myl to each well, and incubate in the dark for 1 hour at RT.
9. Remove the second step reagent, and wash the wells three times with 100 myl of 1× PBS.
10. Remove the PBS, and counter-stain the nuclei by adding 200 myl per well of 2 myg/ml Hoechst 33342 in 1× PBS to each well at least 15 minutes before imaging.
11. View and analyze the cells on an appropriate imaging instrument.
Comment

Related Products: ABIN968537, ABIN967389

Restrictions For Research Use only
Format Liquid
Concentration 0.5 mg/ml
Buffer Aqueous buffered solution.
Preservative Sodium azide
Precaution of Use This product contains sodium azide: a POISONOUS AND HAZARDOUS SUBSTANCE which should be handled by trained staff only.
Storage 4 °C
Supplier Images
anti-Transportin (N-Term) antibody Western blot analysis of transportin in human T leukemia. Jurkat cell lysate (ABIN968537) was probed with Mouse anti-transportin monoclonal antibody at concentrations of 0. 125 (lane 1), 0.0625 (lane 2), and 0.03125 myg/ml (lane 3). Transportin is identified as a band of 95 - 101 kDa.
anti-Transportin (N-Term) antibody (2) Immunofluorescent staining of HeLa (ATCC CCL-2) cells. Cells were seeded in a 96-well imaging plate at ~10,000 cells per well. After overnight incubation, cells were stained using the alcohol perm protocol and the Mouse anti-Transportin monoclonal antibody. The second-step reagent was Alexa Fluor® 555 goat anti-mouse Ig (Invitrogen). Images were captured on a BD Pathway™ 855 Bioimager using a 20x objective. This antibody also stained A549 (ATCC CCL-185) and U-2 OS (ATCC HTB-96) cells and worked with both the Triton™ X-100 and alcohol perm protocols.
Product cited in: Pollard, Michael, Nakielny et al.: "A novel receptor-mediated nuclear protein import pathway." in: Cell, Vol. 86, Issue 6, pp. 985-94, 1996 (PubMed).

Ribbeck, Kutay, Paraskeva et al.: "The translocation of transportin-cargo complexes through nuclear pores is independent of both Ran and energy." in: Current biology : CB, Vol. 9, Issue 1, pp. 47-50, 1999 (PubMed).

Mosammaparast, Pemberton: "Karyopherins: from nuclear-transport mediators to nuclear-function regulators." in: Trends in cell biology, Vol. 14, Issue 10, pp. 547-56, 2004 (PubMed).

Poon, Jans: "Regulation of nuclear transport: central role in development and transformation?" in: Traffic (Copenhagen, Denmark), Vol. 6, Issue 3, pp. 173-86, 2005 (PubMed).

Validation Images
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