Immunofluorescence analysis of Ubiquilin / Dsk2 was performed using 70% confluent log phase SH-SY5Y 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 Ubiquilin / Dsk2 (3D5E2) Mouse Monoclonal Antibody (Product # 37-7700) at 2µg/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 conjµgate (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 (Product # S36938). F-actin (Panel c: red) was stained with Alexa Fluor® 555 Rhodamine Phalloidin (Product # R415, 1:300). Panel d represents the merged image showing nuclear localization. Panel e shows the no primary antibody control. The images were captured at 60X magnification.
|Tested species reactivity||Human, Mouse|
|Published species reactivity||Yeast, Mouse, Human|
|Host / Isotype||Mouse / IgG2b, kappa|
|Immunogen||Recombinant protein derived from an internal region of human ubiquilin.|
|Storage buffer||PBS, pH 7.4|
|Contains||0.1% sodium azide|
|Tested Applications||Dilution *|
|ELISA (ELISA)||Assay Dependent|
|Flow Cytometry (Flow)||3-5 µg/10^6 cells|
|Immunocytochemistry (ICC)||2 µg/ml|
|Immunofluorescence (IF)||2 µg/ml|
|Immunohistochemistry (Paraffin) (IHC (P))||1:20-1:100|
|Immunoprecipitation (IP)||Assay Dependent|
|Western Blot (WB)||1-3 µg/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.
Dsk2 increases the half-life of proteins destined to be degraded by the proteasome, and may modulate proteasome-mediated protein degradation. The Dsk2 protein binds UBE3A and BTRC, and interacts with the 19S proteasome subunit. In the cytoplasm, Dsk2 colocalizes with the proteasome; it is also associated with fibers in mitotic cells in the nucleus. Dsk2 is highly expressed in mitotic cells from metaphase to telophase, while expression in non-mitotic cells is very low.
For Research Use Only. Not for use in diagnostic procedures. Not for resale without express authorization.
Signature changes in ubiquilin expression in the R6/2 mouse model of Huntington's disease.
37-7700 was used in immunohistochemistry and western blot to evaluate the diagnostic significance of ubiquilin expression in Huntington disease
|Safren N,Chang L,Dziki KM,Monteiro MJ||Brain research (1597:37)||2015|
Nuclear proteasomes carry a constitutive posttranslational modification which derails SDS-PAGE (but not CTAB-PAGE).
37-7700 was used in western blot to identify the unique SDS-PAGE pattern of nuclear proteasomes.
|Pitcher DS,de Mattos-Shipley K,Wang Z,Tzortzis K,Goudevenou K,Flynn H,Bohn G,Rahemtulla A,Roberts I,Snijders AP,Karadimitris A,Kleijnen MF||Biochimica et biophysica acta (1844:2222)||2014|
|Yeast||Not Cited||Studies of the role of ubiquitination in the interaction of ubiquilin with the loop and carboxyl terminal regions of presenilin-2.||Ford DL,Monteiro MJ||Biochemistry (46:8827)||2007|
|Human||Not Cited||Ubiquilin 1 modulates amyloid precursor protein trafficking and Abeta secretion.||Hiltunen M,Lu A,Thomas AV,Romano DM,Kim M,Jones PB,Xie Z,Kounnas MZ,Wagner SL,Berezovska O,Hyman BT,Tesco G,Bertram L,Tanzi RE||The Journal of biological chemistry (281:32240)||2006|
Ubiquilin-1 overexpression increases the lifespan and delays accumulation of Huntingtin aggregates in the R6/2 mouse model of Huntington's disease.
37-7700 was used in immunohistochemistry and western blot to test if overexpression of ubiquilin-1 delays or prevents neurodegeneration in R6/2 mice
|Safren N,El Ayadi A,Chang L,Terrillion CE,Gould TD,Boehning DF,Monteiro MJ||PloS one (9:null)||2014|
||Studies of the role of ubiquitination in the interaction of ubiquilin with the loop and carboxyl terminal regions of presenilin-2.||Ford DL,Monteiro MJ||Biochemistry (46:8827)||2007|
||Ubiquilin 1 modulates amyloid precursor protein trafficking and Abeta secretion.||Hiltunen M,Lu A,Thomas AV,Romano DM,Kim M,Jones PB,Xie Z,Kounnas MZ,Wagner SL,Berezovska O,Hyman BT,Tesco G,Bertram L,Tanzi RE||The Journal of biological chemistry (281:32240)||2006|