sag (Proteintech)
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93
Structured Review
Proteintech
sag
Sag, supplied by Proteintech, used in various techniques. Bioz Stars score: 93/100, based on 9 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
https://www.bioz.com/result/sag/product/Proteintech
Average 93 stars, based on 9 article reviews
Sag, supplied by Proteintech, used in various techniques. Bioz Stars score: 93/100, based on 9 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
https://www.bioz.com/result/sag/product/Proteintech
Average 93 stars, based on 9 article reviews
sag - by Bioz Stars,
2026-03
93/100 stars
Images
1) Product Images from "The Crosstalk Between CRL5 and APC/C E3 Ligases Regulates Metastasis and Chemosensitivity of Cancer Cells"
Article Title: The Crosstalk Between CRL5 and APC/C E3 Ligases Regulates Metastasis and Chemosensitivity of Cancer Cells
Journal: Advanced Science
doi: 10.1002/advs.202512652
Figure Legend Snippet: APC11 selectively interacts with CUL5 among cullin family members. a) Silver staining pattern of CUL5‐interacting proteins. MIA PaCa‐2 cells stably overexpressing SBP‐S‐CUL5 or mock vector pLVX were harvested for tandem affinity purification, followed by SDS‐PAGE and silver staining (top). APC11 was a candidate CUL5‐binding protein identified by LC‐MS/MS analysis (bottom, scatter plot of CUL5‐binding proteins). b) Co‐immunoprecipitaion (Co‐IP) of exogenous FLAG‐CUL5 and endogenous APC11 in MIA PaCa‐2 cells. c) Co‐IP of exogenous FLAG‐tagged cullin proteins and endogenous APC11 or SAG in HEK293 cells. d) The interaction of exogenous FLAG‐APC11 with endogenous CUL5 in HEK293 cells. e) Co‐IP of endogenous CUL5 and APC11 in MIA PaCa‐2, A549, Hep3B, and PLC/PRF/5 cells. f) CUL5 binds to APC11 in vitro. Recombinant HIS‐APC11 was incubated with purified FLAG‐CUL5 or vector in vitro, followed by FLAG‐pull‐down and then IB with anti‐APC11 and FLAG Abs. WCE, whole cell extracts.
Techniques Used: Silver Staining, Stable Transfection, Plasmid Preparation, Affinity Purification, SDS Page, Binding Assay, Liquid Chromatography with Mass Spectroscopy, Co-Immunoprecipitation Assay, In Vitro, Recombinant, Incubation, Purification
Figure Legend Snippet: APC11 binds to CUL5 in a manner distinct from SAG. a,b) Co‐IP of transfected FLAG‐APC11 (a) or FLAG‐SAG (b) and endogenous CUL5 in HEK293 cells with 1 µM MLN4924 treatment for 12 h. c) The interaction of endogenous CUL5 with APC11 or SAG upon SAG or APC11 knockdown. A549 and PLC/PRF/5 cells were transfected with siRNA oligos targeting APC11, SAG or scrambled control siRNA for 72 h and then immunoprecipitated with IgG or anti‐CUL5 Ab. d) Co‐IP of endogenous APC11, SAG or CAND1 and FLAG‐tagged CUL5 truncations including wild‐type CUL5 (aa 1–780), CUL5‐N (aa 1–400), CUL5‐C (aa 385–780), and CUL5‐ΔSAG (deletion of aa 565‒582) in HEK293 cells. e) A structural modeling illustration of docking APC11 (red) to CUL5 (blue). The putative CUL5‐interaction sites in APC11 are shown in the enlarged image. f) Co‐IP of transfected FLAG‐APC11 wild‐type or mutants in N terminus (including V3A, I5A, W8A, and W14A) and endogenous CUL5 or APC2, in HEK293 cells. g) Schematic representation of the key APC11 residues involved in binding to full‐length CUL5.
Techniques Used: Co-Immunoprecipitation Assay, Transfection, Knockdown, Control, Immunoprecipitation, Binding Assay
Figure Legend Snippet: APC11 depletion promotes CUL5 neddylation but stabilizes integrin β1. a) Immunoblot of indicated proteins in PLC/PRF/5, Hep3B, and PANC‐1 cells transfected with siRNA targeting APC11 or with a scrambled control siRNA. b) Co‐IP of transfected FLAG‐CUL5 and endogenous CAND1, COPS5, UBE2F or SAG in HEK293 cells transfected with indicated siRNA oligos. c) Co‐IP of endogenous CUL5 with UBE2F or APC11 in A549 cells transfected with indicated siRNA oligos. d) Immunoblot of CUL5 in A549 and PLC/PRF/5 cells transfected with indicated siRNA oligos. e) The stability of SOCS3 and integrin β1 in A549 and MIA PaCa‐2 cells upon APC11 knockdown. A549 and MIA PaCa‐2 cells were transfected with indicated siRNA oligos for 72 h and then treated with 100 µg mL −1 CHX for the indicated time periods, followed by IB analysis. Densitometry quantifications were performed with ImageJ, and the decay curves are shown (right). Data are presented as mean ± SEM, n = 3. For statistical analysis, significances were determined by Student's t ‐test. * p < 0.05, ** p < 0.01, *** p < 0.001. f) Diagram illustrating how APC11 depletion promotes CUL5 neddylation and leads to integrin β1 accumulation.
Techniques Used: Western Blot, Transfection, Control, Co-Immunoprecipitation Assay, Knockdown
