halo-protac3 Search Results


haloprotac3 ligand  (Promega)
90
Promega haloprotac3 ligand
Schematic of <t>HaloPROTAC3</t> degradation of HaloTag protein fusions in live cells. First, CRISPR/Cas9 technology is used to insert HaloTag or HiBiT‐HaloTag into the genomic locus of the target protein using a dsDNA donor plasmid and Cas9‐crRNA complex (1). After the HaloTag fusion is expressed, cells are treated with HaloPROTAC3(2). HaloPROTAC3 induces a ternary complex between the VHL E3 ligase component and HaloTag protein fusion, resulting degradation of the HaloTag target protein via the ubiquitin‐proteasomal pathway (3).
Haloprotac3 Ligand, supplied by Promega, used in various techniques. Bioz Stars score: 90/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
https://www.bioz.com/product/halo-protac3/pmc07818660-64-34-37?v=Promega
Average 90 stars, based on 1 article reviews
haloprotac3 ligand - by Bioz Stars, 2026-08
90/100 stars
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haloprotac3-mediated degradation assay in primary human hepatocytes  (BioIVT Inc)
90
BioIVT Inc haloprotac3-mediated degradation assay in primary human hepatocytes
Schematic of <t>HaloPROTAC3</t> degradation of HaloTag protein fusions in live cells. First, CRISPR/Cas9 technology is used to insert HaloTag or HiBiT‐HaloTag into the genomic locus of the target protein using a dsDNA donor plasmid and Cas9‐crRNA complex (1). After the HaloTag fusion is expressed, cells are treated with HaloPROTAC3(2). HaloPROTAC3 induces a ternary complex between the VHL E3 ligase component and HaloTag protein fusion, resulting degradation of the HaloTag target protein via the ubiquitin‐proteasomal pathway (3).
Haloprotac3 Mediated Degradation Assay In Primary Human Hepatocytes, supplied by BioIVT Inc, used in various techniques. Bioz Stars score: 90/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
https://www.bioz.com/product/halo-protac3/pm40419780-285-5-10?v=BioIVT+Inc
Average 90 stars, based on 1 article reviews
haloprotac3-mediated degradation assay in primary human hepatocytes - by Bioz Stars, 2026-08
90/100 stars
  Buy from Supplier
haloprotac3  (Biosynth Carbosynth)
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haloprotac3  (TargetMol)
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Image Search Results


Schematic of HaloPROTAC3 degradation of HaloTag protein fusions in live cells. First, CRISPR/Cas9 technology is used to insert HaloTag or HiBiT‐HaloTag into the genomic locus of the target protein using a dsDNA donor plasmid and Cas9‐crRNA complex (1). After the HaloTag fusion is expressed, cells are treated with HaloPROTAC3(2). HaloPROTAC3 induces a ternary complex between the VHL E3 ligase component and HaloTag protein fusion, resulting degradation of the HaloTag target protein via the ubiquitin‐proteasomal pathway (3).

Journal: Current Protocols in Pharmacology

Article Title: Targeted Protein Degradation Phenotypic Studies Using HaloTag CRISPR/Cas9 Endogenous Tagging Coupled with HaloPROTAC3

doi: 10.1002/cpph.81

Figure Lengend Snippet: Schematic of HaloPROTAC3 degradation of HaloTag protein fusions in live cells. First, CRISPR/Cas9 technology is used to insert HaloTag or HiBiT‐HaloTag into the genomic locus of the target protein using a dsDNA donor plasmid and Cas9‐crRNA complex (1). After the HaloTag fusion is expressed, cells are treated with HaloPROTAC3(2). HaloPROTAC3 induces a ternary complex between the VHL E3 ligase component and HaloTag protein fusion, resulting degradation of the HaloTag target protein via the ubiquitin‐proteasomal pathway (3).

Article Snippet: HaloTag or HiBiT‐HaloTag CRIPSR/Cas9‐edited cells DPBS (Gibco, cat. no. 14190‐144) 0.05% trypsin/EDTA (Invitrogen, cat. no. 25300‐054) or 0.25% trypsin/EDTA (Invitrogen, cat. no 25200‐056), depending on cell line Complete growth medium for cell type of choice HaloPROTAC3 ligand (Promega, cat. no. GA3110) ent ‐HaloPROTAC3 ligand (Promega, cat. no. GA4110) Dimethyl sulfoxide (DMSO; Sigma, cat. no. 02660‐100 ml) Optional materials for live‐cell kinetic degradation of HiBiT‐HaloTag fusions: LgBiT Expression Vector (Promega, cat. no. N2681) Nano‐Glo Endurazine Live Cell Substrate (Promega, cat. no. N2570/1/2) CO 2 ‐independent medium (Gibco, cat. no. 18045‐088) FBS (VWR, cat. no. 89510‐194) Six‐well plates Appropriate incubator for cell line Dilution reservoirs (Dilux, cat. no. D‐1002) White 96‐well plates (Corning, cat. no. 3917) Optional material for live‐cell kinetic degradation of HiBiT‐HaloTag fusions: Luminometer such as the GloMax Discover Microplate Reader (Promega, cat. no. GM3000) or CLARIOstar Plus (BMG Labtech)

Techniques: CRISPR, Plasmid Preparation

Endpoint analysis of a homozygous β‐catenin‐HaloTag‐HiBiT endogenous protein fusion in HEK293 cells that stably express LgBiT after degradation with HaloPROTAC3. Cells contained the endogenous β‐catenin‐HaloTag‐HiBiT protein fusion were treated with HaloPROTAC3 for 3 or 24 hr, and degradation was detected with an endpoint HiBiT lytic assay ( A ). HaloPROTAC3 caused rapid reduction in the amount of tagged protein in the cell, which was visualized through luminescence imaging on an Olympus LV200 microscope ( B ). Phenotypic characterization was performed on cells that were treated with HaloPROTAC3. Cells containing the endogenous β‐catenin‐HaloTag‐HiBiT protein fusion were transfected with a TCF firefly luciferase reporter before being treating with mWnt3a. HaloPROTAC3 was used to degrade the beta‐catenin‐HaloTag‐HiBiT protein fusion, and after 24 hr a lytic dual luciferase assay was performed to measure firefly luciferase expressed from the TCF reporter ( C ) and β‐catenin HiBiT luminescence ( D ). TCF expression was not induced in cells that were treated with HaloPROTAC3 because β‐catenin‐HaloTag‐HiBiT protein fusions were degraded and could not enter the nucleus upon treatment with mWnt3a.

Journal: Current Protocols in Pharmacology

Article Title: Targeted Protein Degradation Phenotypic Studies Using HaloTag CRISPR/Cas9 Endogenous Tagging Coupled with HaloPROTAC3

doi: 10.1002/cpph.81

Figure Lengend Snippet: Endpoint analysis of a homozygous β‐catenin‐HaloTag‐HiBiT endogenous protein fusion in HEK293 cells that stably express LgBiT after degradation with HaloPROTAC3. Cells contained the endogenous β‐catenin‐HaloTag‐HiBiT protein fusion were treated with HaloPROTAC3 for 3 or 24 hr, and degradation was detected with an endpoint HiBiT lytic assay ( A ). HaloPROTAC3 caused rapid reduction in the amount of tagged protein in the cell, which was visualized through luminescence imaging on an Olympus LV200 microscope ( B ). Phenotypic characterization was performed on cells that were treated with HaloPROTAC3. Cells containing the endogenous β‐catenin‐HaloTag‐HiBiT protein fusion were transfected with a TCF firefly luciferase reporter before being treating with mWnt3a. HaloPROTAC3 was used to degrade the beta‐catenin‐HaloTag‐HiBiT protein fusion, and after 24 hr a lytic dual luciferase assay was performed to measure firefly luciferase expressed from the TCF reporter ( C ) and β‐catenin HiBiT luminescence ( D ). TCF expression was not induced in cells that were treated with HaloPROTAC3 because β‐catenin‐HaloTag‐HiBiT protein fusions were degraded and could not enter the nucleus upon treatment with mWnt3a.

Article Snippet: HaloTag or HiBiT‐HaloTag CRIPSR/Cas9‐edited cells DPBS (Gibco, cat. no. 14190‐144) 0.05% trypsin/EDTA (Invitrogen, cat. no. 25300‐054) or 0.25% trypsin/EDTA (Invitrogen, cat. no 25200‐056), depending on cell line Complete growth medium for cell type of choice HaloPROTAC3 ligand (Promega, cat. no. GA3110) ent ‐HaloPROTAC3 ligand (Promega, cat. no. GA4110) Dimethyl sulfoxide (DMSO; Sigma, cat. no. 02660‐100 ml) Optional materials for live‐cell kinetic degradation of HiBiT‐HaloTag fusions: LgBiT Expression Vector (Promega, cat. no. N2681) Nano‐Glo Endurazine Live Cell Substrate (Promega, cat. no. N2570/1/2) CO 2 ‐independent medium (Gibco, cat. no. 18045‐088) FBS (VWR, cat. no. 89510‐194) Six‐well plates Appropriate incubator for cell line Dilution reservoirs (Dilux, cat. no. D‐1002) White 96‐well plates (Corning, cat. no. 3917) Optional material for live‐cell kinetic degradation of HiBiT‐HaloTag fusions: Luminometer such as the GloMax Discover Microplate Reader (Promega, cat. no. GM3000) or CLARIOstar Plus (BMG Labtech)

Techniques: Stable Transfection, Imaging, Microscopy, Transfection, Luciferase, Expressing

Kinetic degradation profiles of endogenous nuclear, mitochondrial membrane, and cytoplasmic HiBiT‐HaloTag protein fusions after treatment with HaloPROTAC3. Cells were treated with a series of dilutions of HaloPROTAC3 ( A , C , E ) or ent ‐HaloPROTAC3 ( B , D , F ), and degradation was followed by luminescence detection in live cells through a kinetic degradation assay for 24 hr. An N‐terminal HiBiT‐HaloTag‐nuclear endogenous protein fusion in HEK293 cells that stably express LgBiT, a C‐terminal HaloTag‐HiBiT mitochondrial membrane endogenous protein fusion in parent HEK293 cells with LgBiT introduced by transient transfection, and a C‐terminal HaloTag‐HiBiT‐cytoplasmic protein fusion in HEK293 cells that stably express LgBiT were used, respectively. No degradation was detected in the ent ‐HaloPROTAC3‐treated samples, confirming that the protein loss is due to a PROTAC‐mediated mechanism. ( G ) Rapid degradation rates were observed in all samples, with >50% degradation occurring in the first 3 hr. Around 80% degradation was achieved with each HiBiT‐HaloTag protein fusion regardless of cellular location ( H ). DC 50 values were 8.1 nM for the mitochondrial membrane and nuclear protein and 18.6 nM for the cytoplasmic protein.

Journal: Current Protocols in Pharmacology

Article Title: Targeted Protein Degradation Phenotypic Studies Using HaloTag CRISPR/Cas9 Endogenous Tagging Coupled with HaloPROTAC3

doi: 10.1002/cpph.81

Figure Lengend Snippet: Kinetic degradation profiles of endogenous nuclear, mitochondrial membrane, and cytoplasmic HiBiT‐HaloTag protein fusions after treatment with HaloPROTAC3. Cells were treated with a series of dilutions of HaloPROTAC3 ( A , C , E ) or ent ‐HaloPROTAC3 ( B , D , F ), and degradation was followed by luminescence detection in live cells through a kinetic degradation assay for 24 hr. An N‐terminal HiBiT‐HaloTag‐nuclear endogenous protein fusion in HEK293 cells that stably express LgBiT, a C‐terminal HaloTag‐HiBiT mitochondrial membrane endogenous protein fusion in parent HEK293 cells with LgBiT introduced by transient transfection, and a C‐terminal HaloTag‐HiBiT‐cytoplasmic protein fusion in HEK293 cells that stably express LgBiT were used, respectively. No degradation was detected in the ent ‐HaloPROTAC3‐treated samples, confirming that the protein loss is due to a PROTAC‐mediated mechanism. ( G ) Rapid degradation rates were observed in all samples, with >50% degradation occurring in the first 3 hr. Around 80% degradation was achieved with each HiBiT‐HaloTag protein fusion regardless of cellular location ( H ). DC 50 values were 8.1 nM for the mitochondrial membrane and nuclear protein and 18.6 nM for the cytoplasmic protein.

Article Snippet: HaloTag or HiBiT‐HaloTag CRIPSR/Cas9‐edited cells DPBS (Gibco, cat. no. 14190‐144) 0.05% trypsin/EDTA (Invitrogen, cat. no. 25300‐054) or 0.25% trypsin/EDTA (Invitrogen, cat. no 25200‐056), depending on cell line Complete growth medium for cell type of choice HaloPROTAC3 ligand (Promega, cat. no. GA3110) ent ‐HaloPROTAC3 ligand (Promega, cat. no. GA4110) Dimethyl sulfoxide (DMSO; Sigma, cat. no. 02660‐100 ml) Optional materials for live‐cell kinetic degradation of HiBiT‐HaloTag fusions: LgBiT Expression Vector (Promega, cat. no. N2681) Nano‐Glo Endurazine Live Cell Substrate (Promega, cat. no. N2570/1/2) CO 2 ‐independent medium (Gibco, cat. no. 18045‐088) FBS (VWR, cat. no. 89510‐194) Six‐well plates Appropriate incubator for cell line Dilution reservoirs (Dilux, cat. no. D‐1002) White 96‐well plates (Corning, cat. no. 3917) Optional material for live‐cell kinetic degradation of HiBiT‐HaloTag fusions: Luminometer such as the GloMax Discover Microplate Reader (Promega, cat. no. GM3000) or CLARIOstar Plus (BMG Labtech)

Techniques: Degradation Assay, Stable Transfection, Transfection

Comparison between a HaloTag insertion with HaloPROTAC3 degradation and an FKBP12 F36V insertion with dTag‐13 degradation. Endogenous EPOP was tagged at the C terminus with either HaloTag‐HiBiT or FKBP12 F36V ‐HiBiT in HEK293 cells. HiBiT was used in combination with both tags to allow the detection of luminescent live cells once LgBiT was introduced by transient transfection. Both HaloPROTAC3 ( A ) and dTag‐13 ( B ) produced rapid and robust degradation of the protein fusion with the ∼80% degradation after 24 hr and similar levels of compound ( C ). However, with the dTag‐13 compound ( B ), a hook effect was detected in which at higher concentrations, the rate slows and degradation decreases.

Journal: Current Protocols in Pharmacology

Article Title: Targeted Protein Degradation Phenotypic Studies Using HaloTag CRISPR/Cas9 Endogenous Tagging Coupled with HaloPROTAC3

doi: 10.1002/cpph.81

Figure Lengend Snippet: Comparison between a HaloTag insertion with HaloPROTAC3 degradation and an FKBP12 F36V insertion with dTag‐13 degradation. Endogenous EPOP was tagged at the C terminus with either HaloTag‐HiBiT or FKBP12 F36V ‐HiBiT in HEK293 cells. HiBiT was used in combination with both tags to allow the detection of luminescent live cells once LgBiT was introduced by transient transfection. Both HaloPROTAC3 ( A ) and dTag‐13 ( B ) produced rapid and robust degradation of the protein fusion with the ∼80% degradation after 24 hr and similar levels of compound ( C ). However, with the dTag‐13 compound ( B ), a hook effect was detected in which at higher concentrations, the rate slows and degradation decreases.

Article Snippet: HaloTag or HiBiT‐HaloTag CRIPSR/Cas9‐edited cells DPBS (Gibco, cat. no. 14190‐144) 0.05% trypsin/EDTA (Invitrogen, cat. no. 25300‐054) or 0.25% trypsin/EDTA (Invitrogen, cat. no 25200‐056), depending on cell line Complete growth medium for cell type of choice HaloPROTAC3 ligand (Promega, cat. no. GA3110) ent ‐HaloPROTAC3 ligand (Promega, cat. no. GA4110) Dimethyl sulfoxide (DMSO; Sigma, cat. no. 02660‐100 ml) Optional materials for live‐cell kinetic degradation of HiBiT‐HaloTag fusions: LgBiT Expression Vector (Promega, cat. no. N2681) Nano‐Glo Endurazine Live Cell Substrate (Promega, cat. no. N2570/1/2) CO 2 ‐independent medium (Gibco, cat. no. 18045‐088) FBS (VWR, cat. no. 89510‐194) Six‐well plates Appropriate incubator for cell line Dilution reservoirs (Dilux, cat. no. D‐1002) White 96‐well plates (Corning, cat. no. 3917) Optional material for live‐cell kinetic degradation of HiBiT‐HaloTag fusions: Luminometer such as the GloMax Discover Microplate Reader (Promega, cat. no. GM3000) or CLARIOstar Plus (BMG Labtech)

Techniques: Transfection, Produced