S5a/Angiocidin Antibody - BSA Free

Western Blot: S5a/Angiocidin Antibody [NBP2-19952] - Immunoblots of parental HCT116 cells and two clonal cell lines (clone 14 and clone 13) generated by CRISPR-mediated truncation of hRpn10. Image collected and cropped by CiteAb from the following publication (nature.com/articles/s41467-020-15073-7), licensed under a CC-BY license.

Immunocytochemistry/Immunofluorescence: Proteasome 19S S5A Antibody [NBP2-19952] - Immunofluorescence analysis of paraformaldehyde-fixed A549, using antibody at 1:200 dilution.

Western Blot: Proteasome 19S S5A Antibody [NBP2-19952] - Sample (30 ug of whole cell lysate) A: Hela 10% SDS PAGE gel, diluted at 1:1000.

Western Blot: 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] - 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 (//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] - 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 (//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] - 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 (//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] - 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 (//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] - 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 (//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] - 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 (//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] - 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 (//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] - 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 ( delta 307–377) 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 (//pubmed.ncbi.nlm.nih.gov/32157086), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

• Reactivity: Hu, Mu, Rt, Po, Bv, Ca, Ch, Pm, Rb, RM, Sh, Xp
• Applications: WB, ICC/IF, KD
• Clonality: Polyclonal
• Host: Rabbit
• Conjugate: Unconjugated
• Format: BSA Free

S5a/Angiocidin Antibody - BSA Free Summary

• Immunogen: Recombinant protein encompassing a sequence within the center region of human S5a/Angiocidin. The exact sequence is proprietary.
• Predicted Species: Mouse (98%), Rat (98%), Porcine (100%), Chimpanzee (100%), Rabbit (96%), Bovine (99%), Rhesus Macaque (100%), Chicken (95%), Canine (98%), Sheep (100%), Xenopus (90%). Backed by our 100% Guarantee.
• Isotype: IgG
• Clonality: Polyclonal
• Host: Rabbit
• Gene: PSMD4
• Purity: Antigen Affinity-purified

Applications/Dilutions

• Dilutions:
  • Immunocytochemistry/ Immunofluorescence 1:100-1:1000
  • Knockdown Validated
  • Western Blot 1:500-1:3000
• Theoretical MW: 41 kDa.
  Disclaimer note: The observed molecular weight of the protein may vary from the listed predicted molecular weight due to post translational modifications, post translation cleavages, relative charges, and other experimental factors.

Reactivity Notes

Xenopus laevis (90%).

Packaging, Storage & Formulations

• Storage: Aliquot and store at -20C or -80C. Avoid freeze-thaw cycles.
• Buffer: 0.1M Tris (pH 7), 0.1M Glycine, 20% Glycerol
• Preservative: 0.01% Thimerosal
• Purity: Antigen Affinity-purified

Alternate Names for S5a/Angiocidin Antibody - BSA Free

• 26S proteasome non-ATPase regulatory subunit 4;AF;AF-1;ASF;MCB1;PSMD4;pUB-R5;Rpn10;S5A
• Angiocidin
• ASF
• Macropain
• Mcb1
• PSMD4
• pUB-R5
• Rpn10
• S5a

Background

The 26S proteasome is a multicatalytic proteinase complex with a highly ordered structure composed of 2 complexes, a 20S core and a 19S regulator. The 20S core is composed of 4 rings of 28 non-identical subunits; 2 rings are composed of 7 alpha subunits and 2 rings are composed of 7 beta subunits. The 19S regulator is composed of a base, which contains 6 ATPase subunits and 2 non-ATPase subunits, and a lid, which contains up to 10 non-ATPase subunits. Proteasomes are distributed throughout eukaryotic cells at a high concentration and cleave peptides in an ATP/ubiquitin-dependent process in a non-lysosomal pathway. An essential function of a modified proteasome, the immunoproteasome, is the processing of class I MHC peptides. This gene encodes one of the non-ATPase subunits of the 19S regulator lid. Pseudogenes have been identified on chromosomes 10 and 21. [provided by RefSeq]

Limitations

This product is for research use only and is not approved for use in humans or in clinical diagnosis. Primary Antibodies are guaranteed for 1 year from date of receipt.

⚠ WARNING: This product can expose you to chemicals including mercury, which is known to the State of California to cause reproductive toxicity with developmental effects.  For more information go to www.P65Warnings.ca.gov.

Publications for S5a/Angiocidin Antibody (NBP2-19952)(2)

Publications using NBP2-19952

• Osei-Amponsa V, Sridharan V, Tandon M et al. Impact of losing hRpn13 Pru or UCHL5 on proteasome clearance of ubiquitinated proteins and RA190 cytotoxicity Mol. Cell. Biol. 2020-07-06 [PMID: 32631902]
• Structure of E3 ligase E6AP with a proteasome-binding site provided by substrate receptor hRpn10 Buel GR, Chen X, Chari R et al. Nat Commun [PMID: 32157086] (KD, WB, Human)

Bioinformatics

• Gene Symbol: PSMD4
• Uniprot:
  • P55036(Human)