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Immunofluorescence analysis of RanGAP-1 was done on 70% confluent log phase HeLa cells. The cells were fixed with 4% paraformaldehyde for 10 minutes, permeabilized with 0.1% Triton™ X-100 for 10 minutes, and blocked with 1% BSA for 1 hour at room temperature. The cells were labeled with RanGAP-1 Mouse Monoclonal Antibody (330800) at 2ug/ml in 0.1% BSA and incubated for 3 hours at room temperature and then labeled with Goat anti-Mouse IgG (H+L) Superclonal™ Secondary Antibody, Alexa Fluor® 488 conjugate (Product # A28175) at a dilution of 1:2000 for 45 minutes at room temperature (Panel a: green). Nuclei (Panel b: blue) were stained with SlowFade® Gold Antifade Mountant with DAPI (S36938). F-actin (Panel c: red) was stained with Alexa Fluor® 555 Rhodamine Phalloidin (Product # R415, 1:300). Panel d is a merged image showing Nuclear membrane localization. Panel e is a no primary antibody control. The images were captured at 60X magnification.
|Tested species reactivity||Human, Mouse, Rat|
|Published species reactivity||Hamster, Human, Mouse|
|Host / Isotype||Mouse / IgG1, kappa|
|Immunogen||A fusion protein consisting of the first 203 amino-terminal residues of the mouse RanGAP-1 protein.|
|Storage buffer||PBS, pH 7.4|
|Contains||0.1% sodium azide|
|Tested Applications||Dilution *|
|ELISA (ELISA)||0.1-1.0 ug/ml|
|Flow Cytometry (Flow)||3-5 µg/10^6 cells|
|Immunofluorescence (IF)||1-2 µg/ml|
|Western Blot (WB)||0.1-1.0 ug/ml|
* 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.
RanGAP1, is a homodimeric 65-kD polypeptide that specifically induces the GTPase activity of RAN, but not of RAS by over 1, 000-fold. RanGAP1 is the immediate antagonist of RCC1, a regulator molecule that keeps RAN in the active, GTP-bound state. The RANGAP1 gene encodes a 587-amino acid polypeptide. The sequence is unrelated to that of GTPase activators for other RAS-related proteins, but is 88% identical to Fug1, the murine homolog of yeast Rna1p. RanGAP1 and RCC1 control RAN-dependent transport between the nucleus and cytoplasm. RanGAP1 is a key regulator of the RAN GTP/GDP cycle.
For Research Use Only. Not for use in diagnostic procedures. Not for resale without express authorization.
|Hamster||Not Cited||A pathway linking oxidative stress and the Ran GTPase system in progeria.||Datta S,Snow CJ,Paschal BM||Molecular biology of the cell (25:1202)||2014|
|Human||1:200||SUMO-1 targets RanGAP1 to kinetochores and mitotic spindles.||Joseph J,Tan SH,Karpova TS,McNally JG,Dasso M||The Journal of cell biology (156:595)||2002|
Down-modulation of nucleoporin RanBP2/Nup358 impaired chromosomal alignment and induced mitotic catastrophe.
33-0800 was used in immunocytochemistry and western blot to study the role of nucleoporin RanBP2/Nup358 in chromosomal alignment and mitotic integrity
|Hashizume C,Kobayashi A,Wong RW||Cell death & disease (4:null)||2013|
Localization of Pom121 to the inner nuclear membrane is required for an early step of interphase nuclear pore complex assembly.
33-0800 was used in immunocytochemistry to investigate the contribution of pore membrane protein-121 to nuclear pore assembly.
|Funakoshi T,Clever M,Watanabe A,Imamoto N||Molecular biology of the cell (22:1058)||2011|
||Mutation of SENP1/SuPr-2 reveals an essential role for desumoylation in mouse development.||Yamaguchi T,Sharma P,Athanasiou M,Kumar A,Yamada S,Kuehn MR||Molecular and cellular biology (25:5171)||2005|
|Mouse||Not Cited||Mutation of SENP1/SuPr-2 reveals an essential role for desumoylation in mouse development.||Yamaguchi T,Sharma P,Athanasiou M,Kumar A,Yamada S,Kuehn MR||Molecular and cellular biology (25:5171)||2005|
Fug1; ran GTPase-activating protein 1; SD; segregation distorter homolog; segregation distortion
C79654; Fug1; KIAA1835; mKIAA1835; RANGAP; RANGAP1; SD