S5a/Angiocidin Antibody

Knockdown Validated: S5a/Angiocidin Antibody [NBP2-19952]

Knockdown Validated: S5a/Angiocidin Antibody [NBP2-19952] - hRpn10 was knocked down in HCT116 cells by four different siRNAs and the cell lysates immunoprobed as indicated. Mock and scrambled control samples are included. b-actin is used as a loading control. Image collected and cropped by CiteAb from the following publication (nature.com/articles/s41467-020-15073-7), licensed under a CC-BY license.

Western Blot: S5a/Angiocidin Antibody [NBP2-19952] -

Western Blot: S5a/Angiocidin Antibody [NBP2-19952] - E6AP levels depend on hRpn10.hRpn10 (a) or E6AP (b) was knocked down in HCT116 cells by four different siRNAs & the cell lysates immunoprobed as indicated. Mock & scrambled control samples are included. beta-actin is used as a loading control in a & d. c Lysates from HCT116 or clone 14 cells expressing myc-hRpn10 constructs were immunoprobed as indicated. d Lysates from HCT116 or clone 13 cells expressing HA-E6AP and/or myc-hRpn10 constructs were immunoprobed as indicated. a–d Antibodies used for immunoprobing are indicated to the left of each panel. Source data are provided as a Source Data file. Image collected & cropped by CiteAb from the following publication (https://pubmed.ncbi.nlm.nih.gov/32157086), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Western Blot: S5a/Angiocidin Antibody [NBP2-19952] -

Western Blot: S5a/Angiocidin Antibody [NBP2-19952] - E6AP levels depend on hRpn10.hRpn10 (a) or E6AP (b) was knocked down in HCT116 cells by four different siRNAs & the cell lysates immunoprobed as indicated. Mock & scrambled control samples are included. beta-actin is used as a loading control in a & d. c Lysates from HCT116 or clone 14 cells expressing myc-hRpn10 constructs were immunoprobed as indicated. d Lysates from HCT116 or clone 13 cells expressing HA-E6AP and/or myc-hRpn10 constructs were immunoprobed as indicated. a–d Antibodies used for immunoprobing are indicated to the left of each panel. Source data are provided as a Source Data file. Image collected & cropped by CiteAb from the following publication (https://pubmed.ncbi.nlm.nih.gov/32157086), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Western Blot: S5a/Angiocidin Antibody [NBP2-19952] -

Western Blot: S5a/Angiocidin Antibody [NBP2-19952] - E6AP levels depend on hRpn10.hRpn10 (a) or E6AP (b) was knocked down in HCT116 cells by four different siRNAs & the cell lysates immunoprobed as indicated. Mock & scrambled control samples are included. beta-actin is used as a loading control in a & d. c Lysates from HCT116 or clone 14 cells expressing myc-hRpn10 constructs were immunoprobed as indicated. d Lysates from HCT116 or clone 13 cells expressing HA-E6AP and/or myc-hRpn10 constructs were immunoprobed as indicated. a–d Antibodies used for immunoprobing are indicated to the left of each panel. Source data are provided as a Source Data file. Image collected & cropped by CiteAb from the following publication (https://pubmed.ncbi.nlm.nih.gov/32157086), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Western Blot: S5a/Angiocidin Antibody [NBP2-19952] -

Western Blot: S5a/Angiocidin Antibody [NBP2-19952] - hRpn10 RAZUL contributes E6AP to the proteasome.a Immunoblots of Rpt3 immunoprecipitates or WCE from HCT116 or clone 13 lysates expressing myc-hRpn10 constructs. An asterisk “*” indicates heavy chain antibody. Cyclophilin B (Cyp B) is used as a loading control for WCE samples in a–c & hRpn2 as a positive control for the immunoprecipitation. IgG controls are included. b, d Immunoblots of Rpt3 or IgG (control) immunoprecipitates or WCE of lysates from HCT116 cells transfected with empty vector (as a control) or myc-hRpn10 RAZUL. c Immunoblots of Rpt3 immunoprecipitates or WCE from lysates of HCT116 cells transfected with a scrambled control or siRNA against E6AP. a–d Antibodies used for immunoprobing are indicated to the left of each panel. e Pull-down assay for a commercially available mixture of His6-tagged, non-cleavable K48-linked Ub2/Ub4 with incubation of human 26S proteasome (lane 6), 26S proteasome with equimolar E6AP (lane 7), or just E6AP (lane 8). E6AP or 26S proteasome was added to Ni-NTA agarose resin as negative controls (lanes 4 & 5). K48-linked Ub2/Ub4, E6AP, & 26S proteasome were loaded directly in lanes 1–3, as indicated. f Selected regions from 1D 13C-edited, 1H NMR experiments acquired at 850 MHz & 25 °C for free 13C-AZUL (black) or mixtures with equimolar unlabeled RAZUL (blue) or 26S proteasome (red). The concentration of each sample was 0.3 μM & 200,000 scans were recorded for each experiment. Source data are provided as a Source Data file. Image collected & cropped by CiteAb from the following publication (https://pubmed.ncbi.nlm.nih.gov/32157086), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Western Blot: S5a/Angiocidin Antibody [NBP2-19952] -

Western Blot: S5a/Angiocidin Antibody [NBP2-19952] - hRpn10 RAZUL contributes E6AP to the proteasome.a Immunoblots of Rpt3 immunoprecipitates or WCE from HCT116 or clone 13 lysates expressing myc-hRpn10 constructs. An asterisk “*” indicates heavy chain antibody. Cyclophilin B (Cyp B) is used as a loading control for WCE samples in a–c & hRpn2 as a positive control for the immunoprecipitation. IgG controls are included. b, d Immunoblots of Rpt3 or IgG (control) immunoprecipitates or WCE of lysates from HCT116 cells transfected with empty vector (as a control) or myc-hRpn10 RAZUL. c Immunoblots of Rpt3 immunoprecipitates or WCE from lysates of HCT116 cells transfected with a scrambled control or siRNA against E6AP. a–d Antibodies used for immunoprobing are indicated to the left of each panel. e Pull-down assay for a commercially available mixture of His6-tagged, non-cleavable K48-linked Ub2/Ub4 with incubation of human 26S proteasome (lane 6), 26S proteasome with equimolar E6AP (lane 7), or just E6AP (lane 8). E6AP or 26S proteasome was added to Ni-NTA agarose resin as negative controls (lanes 4 & 5). K48-linked Ub2/Ub4, E6AP, & 26S proteasome were loaded directly in lanes 1–3, as indicated. f Selected regions from 1D 13C-edited, 1H NMR experiments acquired at 850 MHz & 25 °C for free 13C-AZUL (black) or mixtures with equimolar unlabeled RAZUL (blue) or 26S proteasome (red). The concentration of each sample was 0.3 μM & 200,000 scans were recorded for each experiment. Source data are provided as a Source Data file. Image collected & cropped by CiteAb from the following publication (https://pubmed.ncbi.nlm.nih.gov/32157086), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Western Blot: S5a/Angiocidin Antibody [NBP2-19952] -

Western Blot: S5a/Angiocidin Antibody [NBP2-19952] - hRpn10 RAZUL contributes E6AP to the proteasome.a Immunoblots of Rpt3 immunoprecipitates or WCE from HCT116 or clone 13 lysates expressing myc-hRpn10 constructs. An asterisk “*” indicates heavy chain antibody. Cyclophilin B (Cyp B) is used as a loading control for WCE samples in a–c & hRpn2 as a positive control for the immunoprecipitation. IgG controls are included. b, d Immunoblots of Rpt3 or IgG (control) immunoprecipitates or WCE of lysates from HCT116 cells transfected with empty vector (as a control) or myc-hRpn10 RAZUL. c Immunoblots of Rpt3 immunoprecipitates or WCE from lysates of HCT116 cells transfected with a scrambled control or siRNA against E6AP. a–d Antibodies used for immunoprobing are indicated to the left of each panel. e Pull-down assay for a commercially available mixture of His6-tagged, non-cleavable K48-linked Ub2/Ub4 with incubation of human 26S proteasome (lane 6), 26S proteasome with equimolar E6AP (lane 7), or just E6AP (lane 8). E6AP or 26S proteasome was added to Ni-NTA agarose resin as negative controls (lanes 4 & 5). K48-linked Ub2/Ub4, E6AP, & 26S proteasome were loaded directly in lanes 1–3, as indicated. f Selected regions from 1D 13C-edited, 1H NMR experiments acquired at 850 MHz & 25 °C for free 13C-AZUL (black) or mixtures with equimolar unlabeled RAZUL (blue) or 26S proteasome (red). The concentration of each sample was 0.3 μM & 200,000 scans were recorded for each experiment. Source data are provided as a Source Data file. Image collected & cropped by CiteAb from the following publication (https://pubmed.ncbi.nlm.nih.gov/32157086), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Western Blot: S5a/Angiocidin Antibody [NBP2-19952] -

Western Blot: S5a/Angiocidin Antibody [NBP2-19952] - hRpn10 RAZUL contributes E6AP to the proteasome.a Immunoblots of Rpt3 immunoprecipitates or WCE from HCT116 or clone 13 lysates expressing myc-hRpn10 constructs. An asterisk “*” indicates heavy chain antibody. Cyclophilin B (Cyp B) is used as a loading control for WCE samples in a–c & hRpn2 as a positive control for the immunoprecipitation. IgG controls are included. b, d Immunoblots of Rpt3 or IgG (control) immunoprecipitates or WCE of lysates from HCT116 cells transfected with empty vector (as a control) or myc-hRpn10 RAZUL. c Immunoblots of Rpt3 immunoprecipitates or WCE from lysates of HCT116 cells transfected with a scrambled control or siRNA against E6AP. a–d Antibodies used for immunoprobing are indicated to the left of each panel. e Pull-down assay for a commercially available mixture of His6-tagged, non-cleavable K48-linked Ub2/Ub4 with incubation of human 26S proteasome (lane 6), 26S proteasome with equimolar E6AP (lane 7), or just E6AP (lane 8). E6AP or 26S proteasome was added to Ni-NTA agarose resin as negative controls (lanes 4 & 5). K48-linked Ub2/Ub4, E6AP, & 26S proteasome were loaded directly in lanes 1–3, as indicated. f Selected regions from 1D 13C-edited, 1H NMR experiments acquired at 850 MHz & 25 °C for free 13C-AZUL (black) or mixtures with equimolar unlabeled RAZUL (blue) or 26S proteasome (red). The concentration of each sample was 0.3 μM & 200,000 scans were recorded for each experiment. Source data are provided as a Source Data file. Image collected & cropped by CiteAb from the following publication (https://pubmed.ncbi.nlm.nih.gov/32157086), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Western Blot: S5a/Angiocidin Antibody [NBP2-19952] -

Western Blot: S5a/Angiocidin Antibody [NBP2-19952] - A C-terminal domain in hRpn10 binds E6AP AZUL.a Positions of known functional domains within hRpn10 (top) & E6AP isoform II (bottom). Question mark “?” indicates hRpn10 uncharacterized region & the E6AP catalytic cysteine C843 is indicated. b Pull-down assay of His-tagged hRpn10full-length (full), hRpn10196–377 or hRpn10196–306 without (−) or with (+) incubation of E6AP. c1H, 15N HSQC spectra of 0.2 mM 15N-hRpn10305–377 (black) & with twofold molar excess unlabeled AZUL (green). Shifted signals are labeled. d Table summarizing Kd, kon, & koff average values with standard deviations for the hRpn10305–377: AZUL interaction measured by ITC and/or SPR. N/A, not applicable. e HCT116 lysates expressing empty vector, myc-hRpn10 full length, or myc-Rpn10 with RAZUL deleted ( delta307–377) were subjected to myc-immunoprecipitation with anti-myc-tag nanobody-coupled agarose. Whole cell extracts (WCE) & myc-immunoprecipitates were immunoprobed with the indicated antibodies. Cyclophilin B (Cyp B) is used as a loading control in e & f. f Lysates from HCT116 cells expressing HA-E6AP & the myc-hRpn10 constructs of e were subjected to HA IP followed by immunoblotting with the indicated antibodies. An asterisk “*” indicates non-specific interaction; double asterisk “**” indicates heavy chain antibody. e–f All antibodies used for immunoprobing are indicated to the left of the images. Note that the hRpn10 & E6AP antibodies recognize both endogenous & exogenously expressed protein, causing these panels to show both tagged & endogenous protein. g Schematic representation highlighting interaction domains of hRpn10 including newly identified RAZUL. Source data are provided as a Source Data file. Image collected & cropped by CiteAb from the following publication (https://pubmed.ncbi.nlm.nih.gov/32157086), licensed under a CC-BY license. Not internally tested by Novus Biologicals.