Importin alpha3 antibody

Overview


	Antigen:	Importin alpha3
	Antibody Type:	Monoclonal
	Application:	Western Blotting (WB)
	Host:	Rat
	Quantity:	100 µg (1 mg/mL)
	Price:	266,00 € (Plus shipping costs and VAT)
	Order number:	ABIN130691
	Availability:	7 to 10 days until shipment

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Additional Information: Importin alpha3 antibody

Antigen	Importin alpha3
Immunogen	This antibody was purified from hybridoma (clone 3D10) supernatant using protein G agarose. This hybridoma was established by fusion of mouse myeloma cell SP2 with BN/Cri rat lymphocyte immunized with human recombinant full-length Importin a3.
Antibody Type	Monoclonal
Isotype	IgG2a »Matching secondary antibodies
Description	Functional nuclear proteins are selectively imported into the nucleus by transport factors such as importins a and ß. Importin a, which mediates the nuclear import of nuclear localization signal (NLS)-containing proteins, is a member of nuclear transport factors. Importin a binds directly NLS and functions as an adapter for accessing the importin ß-dependent import pathway. The import complex is translocated through the nuclear pore complex by interactions of importin ß with a series of nucleoporins.
Clone	3D10
Host	Rat
Specificity	This antibody reacts with Importin a3 (60 kDa) on Western blotting.

Application Details: Importin alpha3 antibody

Application	Western Blotting (WB)
Application Notes	SDS-PAGE & Western Blotting 1) Wash the cells 3 times with PBS and suspend with 10 volumes of cold Lysis buffer (50 mM Tris-HCl, pH 7.2, 250 mM NaCl, 0.1% NP-40, 2 mM EDTA, 10% glycerol) containing appropriate protease inhibitors. Incubate it at 4 o C with rotating for 30 minutes, then sonicate briefly W estern blot analysis o f Importin a3 expression in Jurkat cells (1 ), HeLa cells (2 ), Raji cells (3 ), H L60 cells (4 ), A 431 cells (5 ), L5178Y cells (6 ), W R 19L cells (7 ), N IH /3T3 cells (8 ), C H O cells (9 ), PC 12 cells (1 0 ) a n d R a t 1 c e lls (1 1 ) u s in g D 169-3. 1 2 3 4 5 6 7 8 9 1 0 1 1 43 67 30 20 KDa (up to 10 seconds). 2) Centrifuge the tube at 12,000 x g for 10 minutes at 4 o C and transfer the supernatant to another tube. Measure the protein concentration of the supernatant and add the cold Lysis buffer to make an 8 mg/mL solution. 3) Mix the sample with an equal volume of Laemmli’s sample buffer. 4) Boil the samples for 2 minutes and centrifuge. Load 10 µL of the sample per lane in a 1 mm thick SDS-polyacrylamide gel for electrophoresis. 5) Blot the protein to a polyvinylidene difluoride (PVDF) membrane at 1 mA/cm 2 for 1 hour in a semi-dry transfer system (Transfer Buffer - 25 mM Tris, 190 mM glycine, 20% MeOH). See the manufacture's manual for specific transfer procedure. 6) To reduce nonspecific binding, soak the membrane in 10% skimmed milk (in PBS, pH 7.2) for 1 hour at room temperature, or overnight at 4 o C. 7) Incubate the membrane with primary antibody diluted with PBS, pH 7.2 containing 1% skimmed milk as suggested in the Applications for 1 hour at room temperature. (The optimal antibody concentration will depend on the experimental conditions.) 8) Wash the membrane with PBS (5 minutes x 6 times). 9) Incubate the membrane with the 1:5,000 HRP conjugated anti-Rat IgG(H+L) (MBL, code IM-0825 ) diluted with 1% skimmed milk (in PBS, pH 7.2) for 1 hour at room temperature. 10) Wash the membrane with PBS (5 minutes x 6 times). 11) Wipe excess buffer from the membrane, then incubate it with appropriate chemiluminescence reagents for 1 minute. Remove extra reagent from the membrane by dabbing with a paper towel, and seal it in plastic wrap. 12) Expose to X-ray film in a dark room for 5 minutes. Develop the film as usual. The optimal conditions for exposure and development may vary. Positive controls for Western blotting, HeLa, Jurkat, Raji, HL60, A431, NIH/3T3, CHO, PC12, Rat1 050218-1 Western blotting: 0.1µg / mL for chemiluminescence detection system. Immunoprecipitation: Not tested. Immunohistochemistry: Not recommended. Flow cytometry: Not tested.
Concentration	1 mg/mL
Buffer	100 µg IgG in 100 µL volume of PBS containing 50% glycerol, pH 7.2. Contains no preservatives.
Restrictions	For Research Use Only

Publications: Importin alpha3 antibody

Publications

Hieda, Tachibana, Fukumoto et al.:
"Nuclear import of the U1A splicesome protein is mediated by importin alpha /beta and Ran in living mammalian cells."
In: The Journal of biological chemistry , Vol. 276, Issue 20, pp. 16824-32, 2001/01 (PubMed)

Sakaguchi, Miyamoto, Yoneda et al.:
"Generation of a rat monoclonal antibody specific for importin alpha3/Qip1."
In: Hybridoma and hybridomics , Vol. 22, Issue 6, pp. 397-400, 2003/01 (PubMed)

Images: Importin alpha3 antibody

Western blot analysis of Importin ?3 expression in Jurkat cells (1), H eLa cells (2), R aji cells (3), H L60 cells (4), A 431 cells (5), L5178Y cells (6), W R 19L cells (7), NIH/3T 3 cells (8), C H O cells (9), P C 12 cells (10) and R at 1 cells (11) using ABIN130691.

Image for Importin alpha3 antibody (1)

External Information: Importin alpha3 antibody

Google Scholar	Find more references for clone 3D10 on Google Scholar™
EMBL Harvester	Search human, mouse or rat genome for antigen Importin alpha3 (Bioinformatic Harvester (EMBL))