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( A ) Recombinant GPLD1-RFP-GFP was mixed with or without ALFA-SAA1 in <t>PBS</t> and preincubated at 37°C for 2 h. Samples were then subjected to immunoprecipitation using ALFA-nanobody beads <t>at</t> <t>4°C.</t> ( B ) Recombinant GPLD1-RFP-GFP was mixed with or without luciferase in PBS and incubated at 42°C for 30 min. Samples were subjected to immunoprecipitation using GFP nanobody beads at 4°C.
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( A ) Recombinant GPLD1-RFP-GFP was mixed with or without ALFA-SAA1 in <t>PBS</t> and preincubated at 37°C for 2 h. Samples were then subjected to immunoprecipitation using ALFA-nanobody beads <t>at</t> <t>4°C.</t> ( B ) Recombinant GPLD1-RFP-GFP was mixed with or without luciferase in PBS and incubated at 42°C for 30 min. Samples were subjected to immunoprecipitation using GFP nanobody beads at 4°C.
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( A ) Recombinant GPLD1-RFP-GFP was mixed with or without ALFA-SAA1 in <t>PBS</t> and preincubated at 37°C for 2 h. Samples were then subjected to immunoprecipitation using ALFA-nanobody beads <t>at</t> <t>4°C.</t> ( B ) Recombinant GPLD1-RFP-GFP was mixed with or without luciferase in PBS and incubated at 42°C for 30 min. Samples were subjected to immunoprecipitation using GFP nanobody beads at 4°C.
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( A ) Recombinant GPLD1-RFP-GFP was mixed with or without ALFA-SAA1 in <t>PBS</t> and preincubated at 37°C for 2 h. Samples were then subjected to immunoprecipitation using ALFA-nanobody beads <t>at</t> <t>4°C.</t> ( B ) Recombinant GPLD1-RFP-GFP was mixed with or without luciferase in PBS and incubated at 42°C for 30 min. Samples were subjected to immunoprecipitation using GFP nanobody beads at 4°C.
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( A ) Recombinant GPLD1-RFP-GFP was mixed with or without ALFA-SAA1 in <t>PBS</t> and preincubated at 37°C for 2 h. Samples were then subjected to immunoprecipitation using ALFA-nanobody beads <t>at</t> <t>4°C.</t> ( B ) Recombinant GPLD1-RFP-GFP was mixed with or without luciferase in PBS and incubated at 42°C for 30 min. Samples were subjected to immunoprecipitation using GFP nanobody beads at 4°C.
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( A ) Recombinant GPLD1-RFP-GFP was mixed with or without ALFA-SAA1 in <t>PBS</t> and preincubated at 37°C for 2 h. Samples were then subjected to immunoprecipitation using ALFA-nanobody beads <t>at</t> <t>4°C.</t> ( B ) Recombinant GPLD1-RFP-GFP was mixed with or without luciferase in PBS and incubated at 42°C for 30 min. Samples were subjected to immunoprecipitation using GFP nanobody beads at 4°C.
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( A ) Recombinant GPLD1-RFP-GFP was mixed with or without ALFA-SAA1 in PBS and preincubated at 37°C for 2 h. Samples were then subjected to immunoprecipitation using ALFA-nanobody beads at 4°C. ( B ) Recombinant GPLD1-RFP-GFP was mixed with or without luciferase in PBS and incubated at 42°C for 30 min. Samples were subjected to immunoprecipitation using GFP nanobody beads at 4°C.

Journal: bioRxiv

Article Title: GPLD1 functions as a scavenger chaperone mediating lysosomal degradation of extracellular aberrant proteins

doi: 10.1101/2025.07.16.665076

Figure Lengend Snippet: ( A ) Recombinant GPLD1-RFP-GFP was mixed with or without ALFA-SAA1 in PBS and preincubated at 37°C for 2 h. Samples were then subjected to immunoprecipitation using ALFA-nanobody beads at 4°C. ( B ) Recombinant GPLD1-RFP-GFP was mixed with or without luciferase in PBS and incubated at 42°C for 30 min. Samples were subjected to immunoprecipitation using GFP nanobody beads at 4°C.

Article Snippet: ALFA-SAA1 (8 μM) was added to 50% bovine plasma (Rockland Immunochemicals, cat# D500-06-0500) diluted in PBS and incubated for 16 h at 4°C.

Techniques: Recombinant, Immunoprecipitation, Luciferase, Incubation