pgl3-basic empty vector Search Results


eef2k  (Sino Biological)
92
Sino Biological eef2k
Eef2k, supplied by Sino Biological, used in various techniques. Bioz Stars score: 92/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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pgl3-basic  (Promega)
90
Promega pgl3-basic
Transcriptional control of CXCR1 by MYT1L and DNA-PK. ( A ) Whole cellular lysates were prepared from BJ-5ta, A-172, M059J, M059K, IMR-5, IMR-32, SH-SY5Y, SK-N-AS, SK-N-BE(2), SK-N-MC, and SK-N-SH cells and subjected to Western blot analysis of MYT1L; GAPDH served as a loading control. ( B ) Total RNA was isolated from BJ-5ta, A-172, M059J, M059K, IMR-5, IMR-32, SH-SY5Y, SK-N-AS, SK-N-BE(2), SK-N-MC, and SK-N-SH cells and subjected to the qRT-PCR analysis of CXCR1 that was performed in triplicate. ( C ) Total RNA was isolated from HEK293-MYT1L, HEK293-GFP, M059J-MYT1L, M059J-GFP, M059K-MYT1L, and M059K-GFP cells and subjected to the qRT-PCR analysis of CXCR1 that was performed in triplicate. ( D , E ) Whole cellular lysates were prepared from HEK293-MYT1L, HEK293-GFP, M059J-MYT1L, M059J-GFP, M059K-MYT1L, and M059K-GFP cells and subjected to Western blot analysis of CXCR1 and CXCR2; actin and GAPDH served as a loading control. Note that the images of blots have been cropped and aligned in the same order as panels “( C )” and “( E )”. ( F ) A diagram of wild-type and mutant CXCR1 promoter/reporter constructs. ( G ) HEK293, M069J, and M059K cells were cotransfected with either an empty vector or <t>pGL3-wtCXCR1-luc</t> or pGL3-mtCXCR1 −2114/−2103 -luc reporter plasmid in combination with pCMV6-MYT1L and pRL-TK; at 24 h after transfection, the luciferase activity was measured in duplicate as described in “Methods”. * indicates p < 0.05.
Pgl3 Basic, 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
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vector control  (Addgene inc)
93
Addgene inc vector control
Transcriptional control of CXCR1 by MYT1L and DNA-PK. ( A ) Whole cellular lysates were prepared from BJ-5ta, A-172, M059J, M059K, IMR-5, IMR-32, SH-SY5Y, SK-N-AS, SK-N-BE(2), SK-N-MC, and SK-N-SH cells and subjected to Western blot analysis of MYT1L; GAPDH served as a loading control. ( B ) Total RNA was isolated from BJ-5ta, A-172, M059J, M059K, IMR-5, IMR-32, SH-SY5Y, SK-N-AS, SK-N-BE(2), SK-N-MC, and SK-N-SH cells and subjected to the qRT-PCR analysis of CXCR1 that was performed in triplicate. ( C ) Total RNA was isolated from HEK293-MYT1L, HEK293-GFP, M059J-MYT1L, M059J-GFP, M059K-MYT1L, and M059K-GFP cells and subjected to the qRT-PCR analysis of CXCR1 that was performed in triplicate. ( D , E ) Whole cellular lysates were prepared from HEK293-MYT1L, HEK293-GFP, M059J-MYT1L, M059J-GFP, M059K-MYT1L, and M059K-GFP cells and subjected to Western blot analysis of CXCR1 and CXCR2; actin and GAPDH served as a loading control. Note that the images of blots have been cropped and aligned in the same order as panels “( C )” and “( E )”. ( F ) A diagram of wild-type and mutant CXCR1 promoter/reporter constructs. ( G ) HEK293, M069J, and M059K cells were cotransfected with either an empty vector or <t>pGL3-wtCXCR1-luc</t> or pGL3-mtCXCR1 −2114/−2103 -luc reporter plasmid in combination with pCMV6-MYT1L and pRL-TK; at 24 h after transfection, the luciferase activity was measured in duplicate as described in “Methods”. * indicates p < 0.05.
Vector Control, supplied by Addgene inc, used in various techniques. Bioz Stars score: 93/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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empty pgl3basic reporter gene vector  (Addgene inc)
93
Addgene inc empty pgl3basic reporter gene vector
Transcriptional control of CXCR1 by MYT1L and DNA-PK. ( A ) Whole cellular lysates were prepared from BJ-5ta, A-172, M059J, M059K, IMR-5, IMR-32, SH-SY5Y, SK-N-AS, SK-N-BE(2), SK-N-MC, and SK-N-SH cells and subjected to Western blot analysis of MYT1L; GAPDH served as a loading control. ( B ) Total RNA was isolated from BJ-5ta, A-172, M059J, M059K, IMR-5, IMR-32, SH-SY5Y, SK-N-AS, SK-N-BE(2), SK-N-MC, and SK-N-SH cells and subjected to the qRT-PCR analysis of CXCR1 that was performed in triplicate. ( C ) Total RNA was isolated from HEK293-MYT1L, HEK293-GFP, M059J-MYT1L, M059J-GFP, M059K-MYT1L, and M059K-GFP cells and subjected to the qRT-PCR analysis of CXCR1 that was performed in triplicate. ( D , E ) Whole cellular lysates were prepared from HEK293-MYT1L, HEK293-GFP, M059J-MYT1L, M059J-GFP, M059K-MYT1L, and M059K-GFP cells and subjected to Western blot analysis of CXCR1 and CXCR2; actin and GAPDH served as a loading control. Note that the images of blots have been cropped and aligned in the same order as panels “( C )” and “( E )”. ( F ) A diagram of wild-type and mutant CXCR1 promoter/reporter constructs. ( G ) HEK293, M069J, and M059K cells were cotransfected with either an empty vector or <t>pGL3-wtCXCR1-luc</t> or pGL3-mtCXCR1 −2114/−2103 -luc reporter plasmid in combination with pCMV6-MYT1L and pRL-TK; at 24 h after transfection, the luciferase activity was measured in duplicate as described in “Methods”. * indicates p < 0.05.
Empty Pgl3basic Reporter Gene Vector, supplied by Addgene inc, used in various techniques. Bioz Stars score: 93/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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empty pgl3 basic plasmids  (Addgene inc)
95
Addgene inc empty pgl3 basic plasmids
Transcriptional control of CXCR1 by MYT1L and DNA-PK. ( A ) Whole cellular lysates were prepared from BJ-5ta, A-172, M059J, M059K, IMR-5, IMR-32, SH-SY5Y, SK-N-AS, SK-N-BE(2), SK-N-MC, and SK-N-SH cells and subjected to Western blot analysis of MYT1L; GAPDH served as a loading control. ( B ) Total RNA was isolated from BJ-5ta, A-172, M059J, M059K, IMR-5, IMR-32, SH-SY5Y, SK-N-AS, SK-N-BE(2), SK-N-MC, and SK-N-SH cells and subjected to the qRT-PCR analysis of CXCR1 that was performed in triplicate. ( C ) Total RNA was isolated from HEK293-MYT1L, HEK293-GFP, M059J-MYT1L, M059J-GFP, M059K-MYT1L, and M059K-GFP cells and subjected to the qRT-PCR analysis of CXCR1 that was performed in triplicate. ( D , E ) Whole cellular lysates were prepared from HEK293-MYT1L, HEK293-GFP, M059J-MYT1L, M059J-GFP, M059K-MYT1L, and M059K-GFP cells and subjected to Western blot analysis of CXCR1 and CXCR2; actin and GAPDH served as a loading control. Note that the images of blots have been cropped and aligned in the same order as panels “( C )” and “( E )”. ( F ) A diagram of wild-type and mutant CXCR1 promoter/reporter constructs. ( G ) HEK293, M069J, and M059K cells were cotransfected with either an empty vector or <t>pGL3-wtCXCR1-luc</t> or pGL3-mtCXCR1 −2114/−2103 -luc reporter plasmid in combination with pCMV6-MYT1L and pRL-TK; at 24 h after transfection, the luciferase activity was measured in duplicate as described in “Methods”. * indicates p < 0.05.
Empty Pgl3 Basic Plasmids, supplied by Addgene inc, used in various techniques. Bioz Stars score: 95/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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luciferase assay system  (Promega)
90
Promega luciferase assay system
Transcriptional control of CXCR1 by MYT1L and DNA-PK. ( A ) Whole cellular lysates were prepared from BJ-5ta, A-172, M059J, M059K, IMR-5, IMR-32, SH-SY5Y, SK-N-AS, SK-N-BE(2), SK-N-MC, and SK-N-SH cells and subjected to Western blot analysis of MYT1L; GAPDH served as a loading control. ( B ) Total RNA was isolated from BJ-5ta, A-172, M059J, M059K, IMR-5, IMR-32, SH-SY5Y, SK-N-AS, SK-N-BE(2), SK-N-MC, and SK-N-SH cells and subjected to the qRT-PCR analysis of CXCR1 that was performed in triplicate. ( C ) Total RNA was isolated from HEK293-MYT1L, HEK293-GFP, M059J-MYT1L, M059J-GFP, M059K-MYT1L, and M059K-GFP cells and subjected to the qRT-PCR analysis of CXCR1 that was performed in triplicate. ( D , E ) Whole cellular lysates were prepared from HEK293-MYT1L, HEK293-GFP, M059J-MYT1L, M059J-GFP, M059K-MYT1L, and M059K-GFP cells and subjected to Western blot analysis of CXCR1 and CXCR2; actin and GAPDH served as a loading control. Note that the images of blots have been cropped and aligned in the same order as panels “( C )” and “( E )”. ( F ) A diagram of wild-type and mutant CXCR1 promoter/reporter constructs. ( G ) HEK293, M069J, and M059K cells were cotransfected with either an empty vector or <t>pGL3-wtCXCR1-luc</t> or pGL3-mtCXCR1 −2114/−2103 -luc reporter plasmid in combination with pCMV6-MYT1L and pRL-TK; at 24 h after transfection, the luciferase activity was measured in duplicate as described in “Methods”. * indicates p < 0.05.
Luciferase Assay System, 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
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pcdna3.1  (Promega)
90
Promega pcdna3.1
Transcriptional control of CXCR1 by MYT1L and DNA-PK. ( A ) Whole cellular lysates were prepared from BJ-5ta, A-172, M059J, M059K, IMR-5, IMR-32, SH-SY5Y, SK-N-AS, SK-N-BE(2), SK-N-MC, and SK-N-SH cells and subjected to Western blot analysis of MYT1L; GAPDH served as a loading control. ( B ) Total RNA was isolated from BJ-5ta, A-172, M059J, M059K, IMR-5, IMR-32, SH-SY5Y, SK-N-AS, SK-N-BE(2), SK-N-MC, and SK-N-SH cells and subjected to the qRT-PCR analysis of CXCR1 that was performed in triplicate. ( C ) Total RNA was isolated from HEK293-MYT1L, HEK293-GFP, M059J-MYT1L, M059J-GFP, M059K-MYT1L, and M059K-GFP cells and subjected to the qRT-PCR analysis of CXCR1 that was performed in triplicate. ( D , E ) Whole cellular lysates were prepared from HEK293-MYT1L, HEK293-GFP, M059J-MYT1L, M059J-GFP, M059K-MYT1L, and M059K-GFP cells and subjected to Western blot analysis of CXCR1 and CXCR2; actin and GAPDH served as a loading control. Note that the images of blots have been cropped and aligned in the same order as panels “( C )” and “( E )”. ( F ) A diagram of wild-type and mutant CXCR1 promoter/reporter constructs. ( G ) HEK293, M069J, and M059K cells were cotransfected with either an empty vector or <t>pGL3-wtCXCR1-luc</t> or pGL3-mtCXCR1 −2114/−2103 -luc reporter plasmid in combination with pCMV6-MYT1L and pRL-TK; at 24 h after transfection, the luciferase activity was measured in duplicate as described in “Methods”. * indicates p < 0.05.
Pcdna3.1, 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
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renilla  (Promega)
90
Promega renilla
Transcriptional control of CXCR1 by MYT1L and DNA-PK. ( A ) Whole cellular lysates were prepared from BJ-5ta, A-172, M059J, M059K, IMR-5, IMR-32, SH-SY5Y, SK-N-AS, SK-N-BE(2), SK-N-MC, and SK-N-SH cells and subjected to Western blot analysis of MYT1L; GAPDH served as a loading control. ( B ) Total RNA was isolated from BJ-5ta, A-172, M059J, M059K, IMR-5, IMR-32, SH-SY5Y, SK-N-AS, SK-N-BE(2), SK-N-MC, and SK-N-SH cells and subjected to the qRT-PCR analysis of CXCR1 that was performed in triplicate. ( C ) Total RNA was isolated from HEK293-MYT1L, HEK293-GFP, M059J-MYT1L, M059J-GFP, M059K-MYT1L, and M059K-GFP cells and subjected to the qRT-PCR analysis of CXCR1 that was performed in triplicate. ( D , E ) Whole cellular lysates were prepared from HEK293-MYT1L, HEK293-GFP, M059J-MYT1L, M059J-GFP, M059K-MYT1L, and M059K-GFP cells and subjected to Western blot analysis of CXCR1 and CXCR2; actin and GAPDH served as a loading control. Note that the images of blots have been cropped and aligned in the same order as panels “( C )” and “( E )”. ( F ) A diagram of wild-type and mutant CXCR1 promoter/reporter constructs. ( G ) HEK293, M069J, and M059K cells were cotransfected with either an empty vector or <t>pGL3-wtCXCR1-luc</t> or pGL3-mtCXCR1 −2114/−2103 -luc reporter plasmid in combination with pCMV6-MYT1L and pRL-TK; at 24 h after transfection, the luciferase activity was measured in duplicate as described in “Methods”. * indicates p < 0.05.
Renilla, 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
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pci-neo  (Promega)
90
Promega pci-neo
Transcriptional control of CXCR1 by MYT1L and DNA-PK. ( A ) Whole cellular lysates were prepared from BJ-5ta, A-172, M059J, M059K, IMR-5, IMR-32, SH-SY5Y, SK-N-AS, SK-N-BE(2), SK-N-MC, and SK-N-SH cells and subjected to Western blot analysis of MYT1L; GAPDH served as a loading control. ( B ) Total RNA was isolated from BJ-5ta, A-172, M059J, M059K, IMR-5, IMR-32, SH-SY5Y, SK-N-AS, SK-N-BE(2), SK-N-MC, and SK-N-SH cells and subjected to the qRT-PCR analysis of CXCR1 that was performed in triplicate. ( C ) Total RNA was isolated from HEK293-MYT1L, HEK293-GFP, M059J-MYT1L, M059J-GFP, M059K-MYT1L, and M059K-GFP cells and subjected to the qRT-PCR analysis of CXCR1 that was performed in triplicate. ( D , E ) Whole cellular lysates were prepared from HEK293-MYT1L, HEK293-GFP, M059J-MYT1L, M059J-GFP, M059K-MYT1L, and M059K-GFP cells and subjected to Western blot analysis of CXCR1 and CXCR2; actin and GAPDH served as a loading control. Note that the images of blots have been cropped and aligned in the same order as panels “( C )” and “( E )”. ( F ) A diagram of wild-type and mutant CXCR1 promoter/reporter constructs. ( G ) HEK293, M069J, and M059K cells were cotransfected with either an empty vector or <t>pGL3-wtCXCR1-luc</t> or pGL3-mtCXCR1 −2114/−2103 -luc reporter plasmid in combination with pCMV6-MYT1L and pRL-TK; at 24 h after transfection, the luciferase activity was measured in duplicate as described in “Methods”. * indicates p < 0.05.
Pci Neo, 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
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pctp promoter plasmids  (Promega)
90
Promega pctp promoter plasmids
Transcriptional control of CXCR1 by MYT1L and DNA-PK. ( A ) Whole cellular lysates were prepared from BJ-5ta, A-172, M059J, M059K, IMR-5, IMR-32, SH-SY5Y, SK-N-AS, SK-N-BE(2), SK-N-MC, and SK-N-SH cells and subjected to Western blot analysis of MYT1L; GAPDH served as a loading control. ( B ) Total RNA was isolated from BJ-5ta, A-172, M059J, M059K, IMR-5, IMR-32, SH-SY5Y, SK-N-AS, SK-N-BE(2), SK-N-MC, and SK-N-SH cells and subjected to the qRT-PCR analysis of CXCR1 that was performed in triplicate. ( C ) Total RNA was isolated from HEK293-MYT1L, HEK293-GFP, M059J-MYT1L, M059J-GFP, M059K-MYT1L, and M059K-GFP cells and subjected to the qRT-PCR analysis of CXCR1 that was performed in triplicate. ( D , E ) Whole cellular lysates were prepared from HEK293-MYT1L, HEK293-GFP, M059J-MYT1L, M059J-GFP, M059K-MYT1L, and M059K-GFP cells and subjected to Western blot analysis of CXCR1 and CXCR2; actin and GAPDH served as a loading control. Note that the images of blots have been cropped and aligned in the same order as panels “( C )” and “( E )”. ( F ) A diagram of wild-type and mutant CXCR1 promoter/reporter constructs. ( G ) HEK293, M069J, and M059K cells were cotransfected with either an empty vector or <t>pGL3-wtCXCR1-luc</t> or pGL3-mtCXCR1 −2114/−2103 -luc reporter plasmid in combination with pCMV6-MYT1L and pRL-TK; at 24 h after transfection, the luciferase activity was measured in duplicate as described in “Methods”. * indicates p < 0.05.
Pctp Promoter Plasmids, 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
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luciferase activities  (Promega)
90
Promega luciferase activities
Transcriptional control of CXCR1 by MYT1L and DNA-PK. ( A ) Whole cellular lysates were prepared from BJ-5ta, A-172, M059J, M059K, IMR-5, IMR-32, SH-SY5Y, SK-N-AS, SK-N-BE(2), SK-N-MC, and SK-N-SH cells and subjected to Western blot analysis of MYT1L; GAPDH served as a loading control. ( B ) Total RNA was isolated from BJ-5ta, A-172, M059J, M059K, IMR-5, IMR-32, SH-SY5Y, SK-N-AS, SK-N-BE(2), SK-N-MC, and SK-N-SH cells and subjected to the qRT-PCR analysis of CXCR1 that was performed in triplicate. ( C ) Total RNA was isolated from HEK293-MYT1L, HEK293-GFP, M059J-MYT1L, M059J-GFP, M059K-MYT1L, and M059K-GFP cells and subjected to the qRT-PCR analysis of CXCR1 that was performed in triplicate. ( D , E ) Whole cellular lysates were prepared from HEK293-MYT1L, HEK293-GFP, M059J-MYT1L, M059J-GFP, M059K-MYT1L, and M059K-GFP cells and subjected to Western blot analysis of CXCR1 and CXCR2; actin and GAPDH served as a loading control. Note that the images of blots have been cropped and aligned in the same order as panels “( C )” and “( E )”. ( F ) A diagram of wild-type and mutant CXCR1 promoter/reporter constructs. ( G ) HEK293, M069J, and M059K cells were cotransfected with either an empty vector or <t>pGL3-wtCXCR1-luc</t> or pGL3-mtCXCR1 −2114/−2103 -luc reporter plasmid in combination with pCMV6-MYT1L and pRL-TK; at 24 h after transfection, the luciferase activity was measured in duplicate as described in “Methods”. * indicates p < 0.05.
Luciferase Activities, 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
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test plasmids  (Promega)
90
Promega test plasmids
Transcriptional control of CXCR1 by MYT1L and DNA-PK. ( A ) Whole cellular lysates were prepared from BJ-5ta, A-172, M059J, M059K, IMR-5, IMR-32, SH-SY5Y, SK-N-AS, SK-N-BE(2), SK-N-MC, and SK-N-SH cells and subjected to Western blot analysis of MYT1L; GAPDH served as a loading control. ( B ) Total RNA was isolated from BJ-5ta, A-172, M059J, M059K, IMR-5, IMR-32, SH-SY5Y, SK-N-AS, SK-N-BE(2), SK-N-MC, and SK-N-SH cells and subjected to the qRT-PCR analysis of CXCR1 that was performed in triplicate. ( C ) Total RNA was isolated from HEK293-MYT1L, HEK293-GFP, M059J-MYT1L, M059J-GFP, M059K-MYT1L, and M059K-GFP cells and subjected to the qRT-PCR analysis of CXCR1 that was performed in triplicate. ( D , E ) Whole cellular lysates were prepared from HEK293-MYT1L, HEK293-GFP, M059J-MYT1L, M059J-GFP, M059K-MYT1L, and M059K-GFP cells and subjected to Western blot analysis of CXCR1 and CXCR2; actin and GAPDH served as a loading control. Note that the images of blots have been cropped and aligned in the same order as panels “( C )” and “( E )”. ( F ) A diagram of wild-type and mutant CXCR1 promoter/reporter constructs. ( G ) HEK293, M069J, and M059K cells were cotransfected with either an empty vector or <t>pGL3-wtCXCR1-luc</t> or pGL3-mtCXCR1 −2114/−2103 -luc reporter plasmid in combination with pCMV6-MYT1L and pRL-TK; at 24 h after transfection, the luciferase activity was measured in duplicate as described in “Methods”. * indicates p < 0.05.
Test Plasmids, 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
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Image Search Results


Transcriptional control of CXCR1 by MYT1L and DNA-PK. ( A ) Whole cellular lysates were prepared from BJ-5ta, A-172, M059J, M059K, IMR-5, IMR-32, SH-SY5Y, SK-N-AS, SK-N-BE(2), SK-N-MC, and SK-N-SH cells and subjected to Western blot analysis of MYT1L; GAPDH served as a loading control. ( B ) Total RNA was isolated from BJ-5ta, A-172, M059J, M059K, IMR-5, IMR-32, SH-SY5Y, SK-N-AS, SK-N-BE(2), SK-N-MC, and SK-N-SH cells and subjected to the qRT-PCR analysis of CXCR1 that was performed in triplicate. ( C ) Total RNA was isolated from HEK293-MYT1L, HEK293-GFP, M059J-MYT1L, M059J-GFP, M059K-MYT1L, and M059K-GFP cells and subjected to the qRT-PCR analysis of CXCR1 that was performed in triplicate. ( D , E ) Whole cellular lysates were prepared from HEK293-MYT1L, HEK293-GFP, M059J-MYT1L, M059J-GFP, M059K-MYT1L, and M059K-GFP cells and subjected to Western blot analysis of CXCR1 and CXCR2; actin and GAPDH served as a loading control. Note that the images of blots have been cropped and aligned in the same order as panels “( C )” and “( E )”. ( F ) A diagram of wild-type and mutant CXCR1 promoter/reporter constructs. ( G ) HEK293, M069J, and M059K cells were cotransfected with either an empty vector or pGL3-wtCXCR1-luc or pGL3-mtCXCR1 −2114/−2103 -luc reporter plasmid in combination with pCMV6-MYT1L and pRL-TK; at 24 h after transfection, the luciferase activity was measured in duplicate as described in “Methods”. * indicates p < 0.05.

Journal: International Journal of Molecular Sciences

Article Title: A Positive Feedback DNA-PK/MYT1L-CXCR1-ERK1/2 Proliferative Signaling Loop in Glioblastoma

doi: 10.3390/ijms26094398

Figure Lengend Snippet: Transcriptional control of CXCR1 by MYT1L and DNA-PK. ( A ) Whole cellular lysates were prepared from BJ-5ta, A-172, M059J, M059K, IMR-5, IMR-32, SH-SY5Y, SK-N-AS, SK-N-BE(2), SK-N-MC, and SK-N-SH cells and subjected to Western blot analysis of MYT1L; GAPDH served as a loading control. ( B ) Total RNA was isolated from BJ-5ta, A-172, M059J, M059K, IMR-5, IMR-32, SH-SY5Y, SK-N-AS, SK-N-BE(2), SK-N-MC, and SK-N-SH cells and subjected to the qRT-PCR analysis of CXCR1 that was performed in triplicate. ( C ) Total RNA was isolated from HEK293-MYT1L, HEK293-GFP, M059J-MYT1L, M059J-GFP, M059K-MYT1L, and M059K-GFP cells and subjected to the qRT-PCR analysis of CXCR1 that was performed in triplicate. ( D , E ) Whole cellular lysates were prepared from HEK293-MYT1L, HEK293-GFP, M059J-MYT1L, M059J-GFP, M059K-MYT1L, and M059K-GFP cells and subjected to Western blot analysis of CXCR1 and CXCR2; actin and GAPDH served as a loading control. Note that the images of blots have been cropped and aligned in the same order as panels “( C )” and “( E )”. ( F ) A diagram of wild-type and mutant CXCR1 promoter/reporter constructs. ( G ) HEK293, M069J, and M059K cells were cotransfected with either an empty vector or pGL3-wtCXCR1-luc or pGL3-mtCXCR1 −2114/−2103 -luc reporter plasmid in combination with pCMV6-MYT1L and pRL-TK; at 24 h after transfection, the luciferase activity was measured in duplicate as described in “Methods”. * indicates p < 0.05.

Article Snippet: HEK293, M059J, and M059K cells grown to 90% confluency in a 6-well plate were transiently cotransfected with 0.5 μg of either pGL3-Basic (an empty vector, Promega) or pGL3-wtCXCR1-luc or pGL3-mtCXCR1 −2114/−2103 -luc in combination with 1.0 μg of pCMV6-MYT1L (MYT1L-GFP, OriGene, Austin, TX, USA) and 5.0 ng of pRL-TK (Promega, Madison, WI, USA) using Lipofectamine 3000 (Invitrogen) according to the manufacturer’s instructions.

Techniques: Control, Western Blot, Isolation, Quantitative RT-PCR, Mutagenesis, Construct, Plasmid Preparation, Transfection, Luciferase, Activity Assay