Journal: Human Molecular Genetics

Article Title: PSMC5 insufficiency and P320R mutation impair proteasome function

doi: 10.1093/hmg/ddae085

Figure Lengend Snippet: Antibodies used in this paper.

Article Snippet: Rabbit anti-PSMD2 , Proteintech , 14748-1-AP.

Techniques:

Journal: The Journal of Biological Chemistry

Article Title: The SH3 domain in the fucosyltransferase FUT8 controls FUT8 activity and localization and is essential for core fucosylation

doi: 10.1074/jbc.RA120.013079

Figure Lengend Snippet: RPN1 was identified as the protein that binds to FUT8 in an SH3 domain–dependent manner. A, FUT8 WT and ΔSH3 expressed in HEK293 FUT8KO cells were immunoprecipitated (IP), and the precipitated proteins were subjected to silver staining. Arrows, co-immunoprecipitated proteins with FUT8 that were identified by proteomics. B, HEK293 FUT8KO cells were transfected with the plasmids for FUT8 WT and ΔSH3, lysed, and subjected to immunoprecipitation with anti-FUT8 (sheep Ab) antibodies. The immunoprecipitated proteins were analyzed by Western blotting with anti-FUT8 (sheep Ab), anti-RPN1, and anti-GAPDH Abs.

Article Snippet: The following antibodies were used: sheep anti-FUT8 (R&D Systems, AF5768), mouse anti-FUT8 (Fujirebio, clone 15C6), rabbit anti-RPN1 (Abcam, ab198508), mouse anti-GAPDH (Merck Millipore, MAB374), mouse anti-N-cadherin (BD Biosciences, 610920), rabbit anti-GM130 (Cell Signaling Technology, 12480), HRP-conjugated anti-mouse IgG (GE Healthcare, NA931V), HRP-conjugated anti-sheep IgG (Abcam, ab6900), HRP-conjugated anti-rabbit IgG (GE Healthcare, NA934V), Alexa546-conjugated anti-sheep IgG (Invitrogen, A21098), and Alexa488-conjugated anti-rabbit IgG (Invitrogen, A21206).

Techniques: Immunoprecipitation, Silver Staining, Transfection, Western Blot

Journal: The Journal of Biological Chemistry

Article Title: The SH3 domain in the fucosyltransferase FUT8 controls FUT8 activity and localization and is essential for core fucosylation

doi: 10.1074/jbc.RA120.013079

Figure Lengend Snippet: Proteomic identification of the proteins that co-precipitated with FUT8

Article Snippet: The following antibodies were used: sheep anti-FUT8 (R&D Systems, AF5768), mouse anti-FUT8 (Fujirebio, clone 15C6), rabbit anti-RPN1 (Abcam, ab198508), mouse anti-GAPDH (Merck Millipore, MAB374), mouse anti-N-cadherin (BD Biosciences, 610920), rabbit anti-GM130 (Cell Signaling Technology, 12480), HRP-conjugated anti-mouse IgG (GE Healthcare, NA931V), HRP-conjugated anti-sheep IgG (Abcam, ab6900), HRP-conjugated anti-rabbit IgG (GE Healthcare, NA934V), Alexa546-conjugated anti-sheep IgG (Invitrogen, A21098), and Alexa488-conjugated anti-rabbit IgG (Invitrogen, A21206).

Techniques:

Journal: The Journal of Biological Chemistry

Article Title: The SH3 domain in the fucosyltransferase FUT8 controls FUT8 activity and localization and is essential for core fucosylation

doi: 10.1074/jbc.RA120.013079

Figure Lengend Snippet: RPN1 positively regulates FUT8 activity. A, HEK293 WT cells were treated with siRPN1 or siControl for 48 h. The cell lysates were analyzed by Western blotting with anti-FUT8 (mouse Ab), anti-RPN1, and anti-GAPDH Abs. B, the lysates of HEK293 WT cells treated with siRPN1 or siControl were incubated with the FUT8 acceptor substrate and GDP-Fuc, and the acceptor substrates and products were separated by HPLC. C, the FUT8-specific activities were calculated by the peak areas in B and shown as the mean ± S.D. (error bars) (n = 3). D, the mRNA expression levels of FUT8, MGAT3, and MGAT5 in HEK293 WT cells treated with siRPN1 were quantified by real-time PCR and shown as the values relative to those in cells treated with siControl (n = 3, mean ± S.D.). The mRNA levels were normalized to the GAPDH levels. *, p < 0.05, Mann–Whitney U test.

Article Snippet: The following antibodies were used: sheep anti-FUT8 (R&D Systems, AF5768), mouse anti-FUT8 (Fujirebio, clone 15C6), rabbit anti-RPN1 (Abcam, ab198508), mouse anti-GAPDH (Merck Millipore, MAB374), mouse anti-N-cadherin (BD Biosciences, 610920), rabbit anti-GM130 (Cell Signaling Technology, 12480), HRP-conjugated anti-mouse IgG (GE Healthcare, NA931V), HRP-conjugated anti-sheep IgG (Abcam, ab6900), HRP-conjugated anti-rabbit IgG (GE Healthcare, NA934V), Alexa546-conjugated anti-sheep IgG (Invitrogen, A21098), and Alexa488-conjugated anti-rabbit IgG (Invitrogen, A21206).

Techniques: Activity Assay, Western Blot, Incubation, Expressing, Real-time Polymerase Chain Reaction, MANN-WHITNEY