other:
Incubation:Article Title: SEL1L-HRD1 interaction is required to form a functional HRD1 ERAD complex
Article Snippet: Protein was separated on SDS-PAGE, followed by electrophoretic transfer to PVDF (Fisher Scientific) membrane. .. The blots were incubated in 2% BSA/Tri-buffered saline tween-20 (TBST) with primary antibodies overnight at 4 °C: anti-HSP90 (Santa Cruz, #sc-13119, 1:5000), anti-SEL1L (home-made, 1:10,000) , anti-HRD1 (Proteintech, #13473-1, 1:2000), anti-OS9 (Abcam, #ab109510, 1:5000), anti-CD147 (Proteintech, #11989-1, 1:3000), anti-IRE1α (Cell Signaling, #3294, 1:2000), anti-ERLEC1 (Abcam, #ab181166, 1:5000), anti-UBE2J1 (Santa Cruz, #sc-377002, 1:3000), anti-DERL2 (gift from Chih-Chi Andrew Hu, 1:1000), anti-BiP/GRP94 (Abcam, #ab21685, 1:5000), anti-PDI (Enzo, #ADI-SPA-890, 1:5000), anti-FLAG (Sigma, #F1804, 1:1000), anti-HA (Sigma, #H3663, 1:5000), anti-Myc (Sigma, #C3956, 1:3000), anti-Pro-Caspase-3 (Cell Signaling, #9662, 1:2000), anti-cleaved-Caspase-3 (Cell Signaling, #9661, 1:1000), anti-Calbindin (Cell Signaling, #2173, 1:5000), anti-PERK (Cell Signaling, #3192, 1:5000), anti-p-PERK (Cell Signaling, #3179, 1:1000), anti-eIF2α (Cell Signaling, #9722, 1:5000), anti-p-eIF2α (Cell Signaling, #9721, 1:1000), anti-VCP (Proteintech, #10736-1-AP, 1:3000), anti-HERP1 (Abcam, #ab150424, 1:3000), anti-FAM8A1 (Proteintech, #24746-1-AP,1:3000), anti-CHOP (Cell Signaling, #2895S, 1:1000), anti-GFAP (Cell Signaling, #3670S, 1:3000), anti-IbaI (Proteintech, #10904-1, 1:3000). .. Membranes were washed with TBST and incubated with secondary antibodies, either HRP conjugated (Bio-Rad, 1:10,000), anti-Rabbit IgG TrueBlot HRP (Rockland, #18-8816-33, 1:500) or anti-Mouse IgG TrueBlot-HRP (Rockland, #18-8817-31, 1:500) at room temperature for 1 h prior to the ECL chemiluminescence detection system (Bio-Rad) development.
Article Title: SEL1L-HRD1 interaction is required to form a functional HRD1 ERAD complex.
Article Snippet: Protein was separated on SDS-PAGE, followed by electrophoretic transfer to PVDF (Fisher Scientific) membrane. .. The blots were incubated in 2% BSA/Tri-buffered saline tween-20 (TBST) with primary antibodies overnight at 4 °C: anti-HSP90 (Santa Cruz, #sc13119, 1:5000), anti-SEL1L (home-made, 1:10,000)44, anti-HRD1 (Proteintech, #13473-1, 1:2000), anti-OS9 (Abcam, #ab109510, 1:5000), anti-CD147 (Proteintech, #11989-1, 1:3000), anti-IRE1α (Cell Signaling, #3294, 1:2000), anti-ERLEC1 (Abcam, #ab181166, 1:5000), anti-UBE2J1 (Santa Cruz, #sc-377002, 1:3000), anti-DERL2 (gift from Chih-Chi Andrew Hu, 1:1000), anti-BiP/GRP94 (Abcam, #ab21685, 1:5000), anti-PDI (Enzo, #ADI-SPA-890, 1:5000), antiFLAG (Sigma, #F1804, 1:1000), anti-HA (Sigma, #H3663, 1:5000), anti-Myc (Sigma, #C3956, 1:3000), anti-Pro-Caspase-3 (Cell Signaling, #9662, 1:2000), anti-cleaved-Caspase-3 (Cell Signaling, #9661, 1:1000), anti-Calbindin (Cell Signaling, #2173, 1:5000), antiPERK (Cell Signaling, #3192, 1:5000), anti-p-PERK (Cell Signaling, #3179, 1:1000), anti-eIF2α (Cell Signaling, #9722, 1:5000), anti-peIF2α (Cell Signaling, #9721, 1:1000), anti-VCP (Proteintech, #10736-1-AP, 1:3000), anti-HERP1 (Abcam, #ab150424, 1:3000), anti-FAM8A1 (Proteintech, #24746-1-AP,1:3000), anti-CHOP (Cell Signaling, #2895S, 1:1000), anti-GFAP (Cell Signaling, #3670S, 1:3000), anti-IbaI (Proteintech, #10904-1, 1:3000). .. Membranes were washed with TBST and incubated with secondary antibodies, either HRP conjugated (Bio-Rad, 1:10,000), anti-Rabbit IgG TrueBlot HRP (Rockland, #18-8816-33, 1:500) or anti-Mouse IgG TrueBlot-HRP (Rockland, #18-8817-31, 1:500) at room temperature for 1 h prior to the ECL chemiluminescence detection system (BioRad) development.
Article Title: Structural basis and pathological implications of the dimeric OS9-SEL1L-HRD1 ERAD Core Complex.
Article Snippet: Protein was separated on SDS-PAGE, followed by electrophoretic transfer to PVDF (Fisher Scientific) membrane. .. The blots were incubated in 2% BSA/Tri-buffered saline tween20 (TBST) with primary antibodies overnight at 4 °C: anti-HSP90 (Santa Cruz, #sc-13119, 1:5000), anti-GAPDH (Proteintech, #60004-1, Nature Communications | (2026) 17:2064 11 1:5000), anti-SEL1L (home-made, 1:10,000)69, anti-HRD1 (Proteintech, #13473-1, 1:2000), anti-OS9 (Abcam, #ab109510, 1:5000), anti-CD147 (Proteintech, #11989-1, 1:3,000), anti-IRE1α (Cell Signaling, #3294, 1:2,000), anti-UBE2J1 (Santa Cruz, #sc-377002, 1:3,000), anti-Derlin-2 (gift, 1:1000), anti-Derlin-1 (gift, 1:1000), anti-FLAG (Sigma, #F1804, 1:1000), anti-HERPUD1 (HERP1) (Abcam, #ab150424, 1:3000), antiFAM8A1 (Proteintech, #24746-1-AP,1:3000), anti-ubiquitin (SantaCruz, #sc-8017, 1:1000), anti-His (Genscript, # A00174, 1:2000), anti-Strep (Sigma, # SAB2702216, 1:1000). .. Membranes were washed with TBST and incubated with secondary antibodies, HRP conjugated (Bio-Rad, 1:10,000) at room temperature for 1 h for ECL chemiluminescence detection system (Bio-Rad) development.
Article Title: Structural basis and pathological implications of the dimeric OS9-SEL1L-HRD1 ERAD Core Complex
Article Snippet: Protein was separated on SDS-PAGE, followed by electrophoretic transfer to PVDF (Fisher Scientific) membrane. .. The blots were incubated in 2% BSA/Tri-buffered saline tween-20 (TBST) with primary antibodies overnight at 4 °C: anti-HSP90 (Santa Cruz, #sc-13119, 1:5000), anti-GAPDH (Proteintech, #60004-1, 1:5000), anti-SEL1L (home-made, 1:10,000) , anti-HRD1 (Proteintech, #13473-1, 1:2000), anti-OS9 (Abcam, #ab109510, 1:5000), anti-CD147 (Proteintech, #11989-1, 1:3,000), anti-IRE1α (Cell Signaling, #3294, 1:2,000), anti-UBE2J1 (Santa Cruz, #sc-377002, 1:3,000), anti-Derlin-2 (gift, 1:1000), anti-Derlin-1 (gift, 1:1000), anti-FLAG (Sigma, #F1804, 1:1000), anti-HERPUD1 (HERP1) (Abcam, #ab150424, 1:3000), anti-FAM8A1 (Proteintech, #24746-1-AP,1:3000), anti-ubiquitin (Santa Cruz, #sc-8017, 1:1000), anti-His (Genscript, # A00174, 1:2000), anti-Strep (Sigma, # SAB2702216, 1:1000). .. Membranes were washed with TBST and incubated with secondary antibodies, HRP conjugated (Bio-Rad, 1:10,000) at room temperature for 1 h for ECL chemiluminescence detection system (Bio-Rad) development.
Article Title: Biallelic Cys141Tyr variant of SEL1L is associated with neurodevelopmental disorders, agammaglobulinemia, and premature death
Article Snippet: .. The blots were incubated in 2% BSA/Tri-buffered saline tween-20 552 (TBST) with primary antibodies overnight at 4°C: anti-HSP90 (Santa Cruz, #sc-13119, 553 1:5,000), anti-SEL1L (home-made against SEL1L 23-205 aa, ref. (49), 1:10,000) , anti-554 SEL1L (Abcam ab78298 against SEL1L 330-400 aa, 1:1000), anti-HRD1 (Proteintech, 555 #13473-1, 1:2,000), anti-OS9 (Abcam, #ab109510, 1:5,000), anti-ERLEC1 (Abcam, 556 #ab181166, 1:5,000), anti-CD147 (Proteintech, #11989-1, 1:3,000), anti-IRE1α (Cell 557 Signaling, #3294, 1:2,000), anti-UBE2J1 (Santa Cruz, #sc-377002, 1:3,000), anti-Ubiquitin 558 (Santa Cruz, P4D1, 1:1000), anti-LC3 (Cell Signaling, #2775), anti-RNF5 (Bethyl, #A303-559 594A, 1:2000), anti-FLAG (Sigma, F1804, 1:1,000), anti-HA (Sigma, H3663, 1:5,000), anti-560 PERK (Cell Signaling, #3192, 1:5000), anti-p-PERK (Cell Signaling, #3179, 1:1000), anti-561 eIF2α (Cell Signaling, #9722, 1:5000), anti-p-eIF2α (Cell Signaling, #9721, 1:1000), anti-BiP 562 (Abcam, #ab21685, 1:5,000), anti-PDI (Enzo, #ADI-SPA-890, 1:5,000). .. Membranes were 563 washed with TBST and incubated with HRP conjugated secondary antibodies (Bio-Rad, 564 1:10,000) at room temperature for 1 hr for ECL chemiluminescence detection system (Bio-565 Rad) development.
Article Title: Biallelic Cys141Tyr variant of SEL1L is associated with neurodevelopmental disorders, agammaglobulinemia, and premature death
Article Snippet: Protein was separated using SDS-PAGE, followed by electrophoretic transfer to PVDF (Fisher Scientific) membrane. .. The blots were incubated in 2% BSA/TBST with the following primary antibodies overnight at 4°C: anti-HSP90 (Santa Cruz Biotechnology Inc., sc-13119, 1:5,000), anti-SEL1L (homemade, against SEL1L, 23–205 aa, ref. , :10,000), anti-SEL1L (Abcam, ab78298, against SEL1L, 330–400 aa, 1:1000), anti-HRD1 (Proteintech, 13473-1, 1:2,000), anti-OS9 (Abcam, ab109510, 1:5,000), anti-ERLEC1 (Abcam, ab181166, 1:5,000), anti-CD147 (Proteintech, 11989-1, 1:3,000), anti-IRE1α (Cell Signaling Technology, 3294, 1:2,000), anti-UBE2J1 (Santa Cruz Biotechnology Inc., sc-377002, 1:3,000), anti-ubiquitin (Santa Cruz Biotechnology Inc., P4D1, 1:1000), anti-LC3 (Cell Signaling Technology, 2775), anti-RNF5 (Bethyl, A303-594A, 1:2000), anti-FLAG (MilliporeSigma, F1804, 1:1,000), anti-HA (MilliporeSigma, H3663, 1:5,000), anti-PERK (Cell Signaling Technology, 3192, 1:5000), anti-–p-PERK (Cell Signaling Technology, 3179, 1:1,000), anti-eIF2α (Cell Signaling Technology, 9722, 1:5000), anti-p-eIF2α (Cell Signaling Technology, 9721, 1:1000), anti-BiP (Abcam, ab21685, 1:5,000), and anti-PDI (Enzo, ADI-SPA-890, 1:5,000). .. Membranes were washed with TBST and incubated with HRP-conjugated secondary antibodies (Bio-Rad, 1:10,000) at room temperature for 1 hour for ECL Chemiluminescence Detection System (Bio-Rad) development.
Saline:Article Title: SEL1L-HRD1 interaction is required to form a functional HRD1 ERAD complex
Article Snippet: Protein was separated on SDS-PAGE, followed by electrophoretic transfer to PVDF (Fisher Scientific) membrane. .. The blots were incubated in 2% BSA/Tri-buffered saline tween-20 (TBST) with primary antibodies overnight at 4 °C: anti-HSP90 (Santa Cruz, #sc-13119, 1:5000), anti-SEL1L (home-made, 1:10,000) , anti-HRD1 (Proteintech, #13473-1, 1:2000), anti-OS9 (Abcam, #ab109510, 1:5000), anti-CD147 (Proteintech, #11989-1, 1:3000), anti-IRE1α (Cell Signaling, #3294, 1:2000), anti-ERLEC1 (Abcam, #ab181166, 1:5000), anti-UBE2J1 (Santa Cruz, #sc-377002, 1:3000), anti-DERL2 (gift from Chih-Chi Andrew Hu, 1:1000), anti-BiP/GRP94 (Abcam, #ab21685, 1:5000), anti-PDI (Enzo, #ADI-SPA-890, 1:5000), anti-FLAG (Sigma, #F1804, 1:1000), anti-HA (Sigma, #H3663, 1:5000), anti-Myc (Sigma, #C3956, 1:3000), anti-Pro-Caspase-3 (Cell Signaling, #9662, 1:2000), anti-cleaved-Caspase-3 (Cell Signaling, #9661, 1:1000), anti-Calbindin (Cell Signaling, #2173, 1:5000), anti-PERK (Cell Signaling, #3192, 1:5000), anti-p-PERK (Cell Signaling, #3179, 1:1000), anti-eIF2α (Cell Signaling, #9722, 1:5000), anti-p-eIF2α (Cell Signaling, #9721, 1:1000), anti-VCP (Proteintech, #10736-1-AP, 1:3000), anti-HERP1 (Abcam, #ab150424, 1:3000), anti-FAM8A1 (Proteintech, #24746-1-AP,1:3000), anti-CHOP (Cell Signaling, #2895S, 1:1000), anti-GFAP (Cell Signaling, #3670S, 1:3000), anti-IbaI (Proteintech, #10904-1, 1:3000). .. Membranes were washed with TBST and incubated with secondary antibodies, either HRP conjugated (Bio-Rad, 1:10,000), anti-Rabbit IgG TrueBlot HRP (Rockland, #18-8816-33, 1:500) or anti-Mouse IgG TrueBlot-HRP (Rockland, #18-8817-31, 1:500) at room temperature for 1 h prior to the ECL chemiluminescence detection system (Bio-Rad) development.
Article Title: SEL1L-HRD1 interaction is required to form a functional HRD1 ERAD complex.
Article Snippet: Protein was separated on SDS-PAGE, followed by electrophoretic transfer to PVDF (Fisher Scientific) membrane. .. The blots were incubated in 2% BSA/Tri-buffered saline tween-20 (TBST) with primary antibodies overnight at 4 °C: anti-HSP90 (Santa Cruz, #sc13119, 1:5000), anti-SEL1L (home-made, 1:10,000)44, anti-HRD1 (Proteintech, #13473-1, 1:2000), anti-OS9 (Abcam, #ab109510, 1:5000), anti-CD147 (Proteintech, #11989-1, 1:3000), anti-IRE1α (Cell Signaling, #3294, 1:2000), anti-ERLEC1 (Abcam, #ab181166, 1:5000), anti-UBE2J1 (Santa Cruz, #sc-377002, 1:3000), anti-DERL2 (gift from Chih-Chi Andrew Hu, 1:1000), anti-BiP/GRP94 (Abcam, #ab21685, 1:5000), anti-PDI (Enzo, #ADI-SPA-890, 1:5000), antiFLAG (Sigma, #F1804, 1:1000), anti-HA (Sigma, #H3663, 1:5000), anti-Myc (Sigma, #C3956, 1:3000), anti-Pro-Caspase-3 (Cell Signaling, #9662, 1:2000), anti-cleaved-Caspase-3 (Cell Signaling, #9661, 1:1000), anti-Calbindin (Cell Signaling, #2173, 1:5000), antiPERK (Cell Signaling, #3192, 1:5000), anti-p-PERK (Cell Signaling, #3179, 1:1000), anti-eIF2α (Cell Signaling, #9722, 1:5000), anti-peIF2α (Cell Signaling, #9721, 1:1000), anti-VCP (Proteintech, #10736-1-AP, 1:3000), anti-HERP1 (Abcam, #ab150424, 1:3000), anti-FAM8A1 (Proteintech, #24746-1-AP,1:3000), anti-CHOP (Cell Signaling, #2895S, 1:1000), anti-GFAP (Cell Signaling, #3670S, 1:3000), anti-IbaI (Proteintech, #10904-1, 1:3000). .. Membranes were washed with TBST and incubated with secondary antibodies, either HRP conjugated (Bio-Rad, 1:10,000), anti-Rabbit IgG TrueBlot HRP (Rockland, #18-8816-33, 1:500) or anti-Mouse IgG TrueBlot-HRP (Rockland, #18-8817-31, 1:500) at room temperature for 1 h prior to the ECL chemiluminescence detection system (BioRad) development.
Article Title: Structural basis and pathological implications of the dimeric OS9-SEL1L-HRD1 ERAD Core Complex.
Article Snippet: Protein was separated on SDS-PAGE, followed by electrophoretic transfer to PVDF (Fisher Scientific) membrane. .. The blots were incubated in 2% BSA/Tri-buffered saline tween20 (TBST) with primary antibodies overnight at 4 °C: anti-HSP90 (Santa Cruz, #sc-13119, 1:5000), anti-GAPDH (Proteintech, #60004-1, Nature Communications | (2026) 17:2064 11 1:5000), anti-SEL1L (home-made, 1:10,000)69, anti-HRD1 (Proteintech, #13473-1, 1:2000), anti-OS9 (Abcam, #ab109510, 1:5000), anti-CD147 (Proteintech, #11989-1, 1:3,000), anti-IRE1α (Cell Signaling, #3294, 1:2,000), anti-UBE2J1 (Santa Cruz, #sc-377002, 1:3,000), anti-Derlin-2 (gift, 1:1000), anti-Derlin-1 (gift, 1:1000), anti-FLAG (Sigma, #F1804, 1:1000), anti-HERPUD1 (HERP1) (Abcam, #ab150424, 1:3000), antiFAM8A1 (Proteintech, #24746-1-AP,1:3000), anti-ubiquitin (SantaCruz, #sc-8017, 1:1000), anti-His (Genscript, # A00174, 1:2000), anti-Strep (Sigma, # SAB2702216, 1:1000). .. Membranes were washed with TBST and incubated with secondary antibodies, HRP conjugated (Bio-Rad, 1:10,000) at room temperature for 1 h for ECL chemiluminescence detection system (Bio-Rad) development.
Article Title: Structural basis and pathological implications of the dimeric OS9-SEL1L-HRD1 ERAD Core Complex
Article Snippet: Protein was separated on SDS-PAGE, followed by electrophoretic transfer to PVDF (Fisher Scientific) membrane. .. The blots were incubated in 2% BSA/Tri-buffered saline tween-20 (TBST) with primary antibodies overnight at 4 °C: anti-HSP90 (Santa Cruz, #sc-13119, 1:5000), anti-GAPDH (Proteintech, #60004-1, 1:5000), anti-SEL1L (home-made, 1:10,000) , anti-HRD1 (Proteintech, #13473-1, 1:2000), anti-OS9 (Abcam, #ab109510, 1:5000), anti-CD147 (Proteintech, #11989-1, 1:3,000), anti-IRE1α (Cell Signaling, #3294, 1:2,000), anti-UBE2J1 (Santa Cruz, #sc-377002, 1:3,000), anti-Derlin-2 (gift, 1:1000), anti-Derlin-1 (gift, 1:1000), anti-FLAG (Sigma, #F1804, 1:1000), anti-HERPUD1 (HERP1) (Abcam, #ab150424, 1:3000), anti-FAM8A1 (Proteintech, #24746-1-AP,1:3000), anti-ubiquitin (Santa Cruz, #sc-8017, 1:1000), anti-His (Genscript, # A00174, 1:2000), anti-Strep (Sigma, # SAB2702216, 1:1000). .. Membranes were washed with TBST and incubated with secondary antibodies, HRP conjugated (Bio-Rad, 1:10,000) at room temperature for 1 h for ECL chemiluminescence detection system (Bio-Rad) development.
Article Title: Biallelic Cys141Tyr variant of SEL1L is associated with neurodevelopmental disorders, agammaglobulinemia, and premature death
Article Snippet: .. The blots were incubated in 2% BSA/Tri-buffered saline tween-20 552 (TBST) with primary antibodies overnight at 4°C: anti-HSP90 (Santa Cruz, #sc-13119, 553 1:5,000), anti-SEL1L (home-made against SEL1L 23-205 aa, ref. (49), 1:10,000) , anti-554 SEL1L (Abcam ab78298 against SEL1L 330-400 aa, 1:1000), anti-HRD1 (Proteintech, 555 #13473-1, 1:2,000), anti-OS9 (Abcam, #ab109510, 1:5,000), anti-ERLEC1 (Abcam, 556 #ab181166, 1:5,000), anti-CD147 (Proteintech, #11989-1, 1:3,000), anti-IRE1α (Cell 557 Signaling, #3294, 1:2,000), anti-UBE2J1 (Santa Cruz, #sc-377002, 1:3,000), anti-Ubiquitin 558 (Santa Cruz, P4D1, 1:1000), anti-LC3 (Cell Signaling, #2775), anti-RNF5 (Bethyl, #A303-559 594A, 1:2000), anti-FLAG (Sigma, F1804, 1:1,000), anti-HA (Sigma, H3663, 1:5,000), anti-560 PERK (Cell Signaling, #3192, 1:5000), anti-p-PERK (Cell Signaling, #3179, 1:1000), anti-561 eIF2α (Cell Signaling, #9722, 1:5000), anti-p-eIF2α (Cell Signaling, #9721, 1:1000), anti-BiP 562 (Abcam, #ab21685, 1:5,000), anti-PDI (Enzo, #ADI-SPA-890, 1:5,000). .. Membranes were 563 washed with TBST and incubated with HRP conjugated secondary antibodies (Bio-Rad, 564 1:10,000) at room temperature for 1 hr for ECL chemiluminescence detection system (Bio-565 Rad) development.
Recombinant:
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