ceramide c6 Search Results


recombinant proteins azithromycin synovo n a c6 nbd ceramide focus biomolecules biotrend  (MedChemExpress)
94
MedChemExpress recombinant proteins azithromycin synovo n a c6 nbd ceramide focus biomolecules biotrend
Recombinant Proteins Azithromycin Synovo N A C6 Nbd Ceramide Focus Biomolecules Biotrend, supplied by MedChemExpress, used in various techniques. Bioz Stars score: 94/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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c6 nbd ceramide 1 phosphate  (Avanti Polar)
94
Avanti Polar c6 nbd ceramide 1 phosphate
(A) Schematic representation of the sequence organization of OsCERK . The sizes of the 5′-UTR (white), exons (black) and 3′-UTR (grey) are indicated by numbers below the boxes. The sizes of introns are shown above. (B) Phylogenetic analysis of CERKs and the related enzymes sphingosine kinase (SPK)s, and diacylglycerol kinase (DAGK)s. The architecture of each gene was obtained from SMART using the architecture analysis function, and conserved domains (letters a–d) are indicated as follows: a, Pleckstrin homology domain (PH domain; accession number SM00233); b, Diacylglycerol kinase catalytic domain (DAGKc; SMART accession number SM00046); c, segments of low compositional complexity determined by the program; d, diacylgycerol kinase catalytic domain (DAGK_cat; Pfam accession number PF00781). Detailed sequences information is described in . AtCERK is ACD5. (C) CLUSTAL W alignment of CERK from various organisms showing the highly conserved CXXXCXXC motif. (D) RT-PCR expression analysis of OsCERK in leaves, stem and roots of 2-week-old seedlings. β-actin was used as a control. (E) CERK activity of rice extracts. Crude plant extracts made from leaves and roots of rice, respectively, were incubated at 40°C for 30 min as described in . The reaction mixture (1 µl) was resolved on a TLC plate. Purified OsCERK recombinant protein was used as the positive control (+CK). For the negative control (−CK) no extract was added. (F) Comparison of recombinant OsCERK and rACD5. The recombinant protein (1.0 µg) was added in 100 µl reaction system, and incubated at 40°C for 30 min. –CK, no recombinant protein was added. Arrows in (E and F) indicated the formed product <t>C6-NBD-ceramide-1-phosphate</t> (C6-NBD-Cer-1P).
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ceramide c6  (Santa Cruz Biotechnology)
93
Santa Cruz Biotechnology ceramide c6
(A) Schematic representation of the sequence organization of OsCERK . The sizes of the 5′-UTR (white), exons (black) and 3′-UTR (grey) are indicated by numbers below the boxes. The sizes of introns are shown above. (B) Phylogenetic analysis of CERKs and the related enzymes sphingosine kinase (SPK)s, and diacylglycerol kinase (DAGK)s. The architecture of each gene was obtained from SMART using the architecture analysis function, and conserved domains (letters a–d) are indicated as follows: a, Pleckstrin homology domain (PH domain; accession number SM00233); b, Diacylglycerol kinase catalytic domain (DAGKc; SMART accession number SM00046); c, segments of low compositional complexity determined by the program; d, diacylgycerol kinase catalytic domain (DAGK_cat; Pfam accession number PF00781). Detailed sequences information is described in . AtCERK is ACD5. (C) CLUSTAL W alignment of CERK from various organisms showing the highly conserved CXXXCXXC motif. (D) RT-PCR expression analysis of OsCERK in leaves, stem and roots of 2-week-old seedlings. β-actin was used as a control. (E) CERK activity of rice extracts. Crude plant extracts made from leaves and roots of rice, respectively, were incubated at 40°C for 30 min as described in . The reaction mixture (1 µl) was resolved on a TLC plate. Purified OsCERK recombinant protein was used as the positive control (+CK). For the negative control (−CK) no extract was added. (F) Comparison of recombinant OsCERK and rACD5. The recombinant protein (1.0 µg) was added in 100 µl reaction system, and incubated at 40°C for 30 min. –CK, no recombinant protein was added. Arrows in (E and F) indicated the formed product <t>C6-NBD-ceramide-1-phosphate</t> (C6-NBD-Cer-1P).
Ceramide C6, supplied by Santa Cruz Biotechnology, 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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c6 ceramide  (Avanti Polar)
95
Avanti Polar c6 ceramide
(A) Schematic representation of the sequence organization of OsCERK . The sizes of the 5′-UTR (white), exons (black) and 3′-UTR (grey) are indicated by numbers below the boxes. The sizes of introns are shown above. (B) Phylogenetic analysis of CERKs and the related enzymes sphingosine kinase (SPK)s, and diacylglycerol kinase (DAGK)s. The architecture of each gene was obtained from SMART using the architecture analysis function, and conserved domains (letters a–d) are indicated as follows: a, Pleckstrin homology domain (PH domain; accession number SM00233); b, Diacylglycerol kinase catalytic domain (DAGKc; SMART accession number SM00046); c, segments of low compositional complexity determined by the program; d, diacylgycerol kinase catalytic domain (DAGK_cat; Pfam accession number PF00781). Detailed sequences information is described in . AtCERK is ACD5. (C) CLUSTAL W alignment of CERK from various organisms showing the highly conserved CXXXCXXC motif. (D) RT-PCR expression analysis of OsCERK in leaves, stem and roots of 2-week-old seedlings. β-actin was used as a control. (E) CERK activity of rice extracts. Crude plant extracts made from leaves and roots of rice, respectively, were incubated at 40°C for 30 min as described in . The reaction mixture (1 µl) was resolved on a TLC plate. Purified OsCERK recombinant protein was used as the positive control (+CK). For the negative control (−CK) no extract was added. (F) Comparison of recombinant OsCERK and rACD5. The recombinant protein (1.0 µg) was added in 100 µl reaction system, and incubated at 40°C for 30 min. –CK, no recombinant protein was added. Arrows in (E and F) indicated the formed product <t>C6-NBD-ceramide-1-phosphate</t> (C6-NBD-Cer-1P).
C6 Ceramide, supplied by Avanti Polar, 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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c6 nbd ceramide  (Santa Cruz Biotechnology)
93
Santa Cruz Biotechnology c6 nbd ceramide
(A) Schematic representation of the sequence organization of OsCERK . The sizes of the 5′-UTR (white), exons (black) and 3′-UTR (grey) are indicated by numbers below the boxes. The sizes of introns are shown above. (B) Phylogenetic analysis of CERKs and the related enzymes sphingosine kinase (SPK)s, and diacylglycerol kinase (DAGK)s. The architecture of each gene was obtained from SMART using the architecture analysis function, and conserved domains (letters a–d) are indicated as follows: a, Pleckstrin homology domain (PH domain; accession number SM00233); b, Diacylglycerol kinase catalytic domain (DAGKc; SMART accession number SM00046); c, segments of low compositional complexity determined by the program; d, diacylgycerol kinase catalytic domain (DAGK_cat; Pfam accession number PF00781). Detailed sequences information is described in . AtCERK is ACD5. (C) CLUSTAL W alignment of CERK from various organisms showing the highly conserved CXXXCXXC motif. (D) RT-PCR expression analysis of OsCERK in leaves, stem and roots of 2-week-old seedlings. β-actin was used as a control. (E) CERK activity of rice extracts. Crude plant extracts made from leaves and roots of rice, respectively, were incubated at 40°C for 30 min as described in . The reaction mixture (1 µl) was resolved on a TLC plate. Purified OsCERK recombinant protein was used as the positive control (+CK). For the negative control (−CK) no extract was added. (F) Comparison of recombinant OsCERK and rACD5. The recombinant protein (1.0 µg) was added in 100 µl reaction system, and incubated at 40°C for 30 min. –CK, no recombinant protein was added. Arrows in (E and F) indicated the formed product <t>C6-NBD-ceramide-1-phosphate</t> (C6-NBD-Cer-1P).
C6 Nbd Ceramide, supplied by Santa Cruz Biotechnology, 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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c6ceramide  (Larodan)
90
Larodan c6ceramide
(A) Schematic representation of the sequence organization of OsCERK . The sizes of the 5′-UTR (white), exons (black) and 3′-UTR (grey) are indicated by numbers below the boxes. The sizes of introns are shown above. (B) Phylogenetic analysis of CERKs and the related enzymes sphingosine kinase (SPK)s, and diacylglycerol kinase (DAGK)s. The architecture of each gene was obtained from SMART using the architecture analysis function, and conserved domains (letters a–d) are indicated as follows: a, Pleckstrin homology domain (PH domain; accession number SM00233); b, Diacylglycerol kinase catalytic domain (DAGKc; SMART accession number SM00046); c, segments of low compositional complexity determined by the program; d, diacylgycerol kinase catalytic domain (DAGK_cat; Pfam accession number PF00781). Detailed sequences information is described in . AtCERK is ACD5. (C) CLUSTAL W alignment of CERK from various organisms showing the highly conserved CXXXCXXC motif. (D) RT-PCR expression analysis of OsCERK in leaves, stem and roots of 2-week-old seedlings. β-actin was used as a control. (E) CERK activity of rice extracts. Crude plant extracts made from leaves and roots of rice, respectively, were incubated at 40°C for 30 min as described in . The reaction mixture (1 µl) was resolved on a TLC plate. Purified OsCERK recombinant protein was used as the positive control (+CK). For the negative control (−CK) no extract was added. (F) Comparison of recombinant OsCERK and rACD5. The recombinant protein (1.0 µg) was added in 100 µl reaction system, and incubated at 40°C for 30 min. –CK, no recombinant protein was added. Arrows in (E and F) indicated the formed product <t>C6-NBD-ceramide-1-phosphate</t> (C6-NBD-Cer-1P).
C6ceramide, supplied by Larodan, 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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p90rsk agonist ceramide c6  (MedChemExpress)
93
MedChemExpress p90rsk agonist ceramide c6
(A) Schematic representation of the sequence organization of OsCERK . The sizes of the 5′-UTR (white), exons (black) and 3′-UTR (grey) are indicated by numbers below the boxes. The sizes of introns are shown above. (B) Phylogenetic analysis of CERKs and the related enzymes sphingosine kinase (SPK)s, and diacylglycerol kinase (DAGK)s. The architecture of each gene was obtained from SMART using the architecture analysis function, and conserved domains (letters a–d) are indicated as follows: a, Pleckstrin homology domain (PH domain; accession number SM00233); b, Diacylglycerol kinase catalytic domain (DAGKc; SMART accession number SM00046); c, segments of low compositional complexity determined by the program; d, diacylgycerol kinase catalytic domain (DAGK_cat; Pfam accession number PF00781). Detailed sequences information is described in . AtCERK is ACD5. (C) CLUSTAL W alignment of CERK from various organisms showing the highly conserved CXXXCXXC motif. (D) RT-PCR expression analysis of OsCERK in leaves, stem and roots of 2-week-old seedlings. β-actin was used as a control. (E) CERK activity of rice extracts. Crude plant extracts made from leaves and roots of rice, respectively, were incubated at 40°C for 30 min as described in . The reaction mixture (1 µl) was resolved on a TLC plate. Purified OsCERK recombinant protein was used as the positive control (+CK). For the negative control (−CK) no extract was added. (F) Comparison of recombinant OsCERK and rACD5. The recombinant protein (1.0 µg) was added in 100 µl reaction system, and incubated at 40°C for 30 min. –CK, no recombinant protein was added. Arrows in (E and F) indicated the formed product <t>C6-NBD-ceramide-1-phosphate</t> (C6-NBD-Cer-1P).
P90rsk Agonist Ceramide C6, supplied by MedChemExpress, 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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nbd c6 ceramide complex  (Biotium)
93
Biotium nbd c6 ceramide complex
(A) Schematic representation of the sequence organization of OsCERK . The sizes of the 5′-UTR (white), exons (black) and 3′-UTR (grey) are indicated by numbers below the boxes. The sizes of introns are shown above. (B) Phylogenetic analysis of CERKs and the related enzymes sphingosine kinase (SPK)s, and diacylglycerol kinase (DAGK)s. The architecture of each gene was obtained from SMART using the architecture analysis function, and conserved domains (letters a–d) are indicated as follows: a, Pleckstrin homology domain (PH domain; accession number SM00233); b, Diacylglycerol kinase catalytic domain (DAGKc; SMART accession number SM00046); c, segments of low compositional complexity determined by the program; d, diacylgycerol kinase catalytic domain (DAGK_cat; Pfam accession number PF00781). Detailed sequences information is described in . AtCERK is ACD5. (C) CLUSTAL W alignment of CERK from various organisms showing the highly conserved CXXXCXXC motif. (D) RT-PCR expression analysis of OsCERK in leaves, stem and roots of 2-week-old seedlings. β-actin was used as a control. (E) CERK activity of rice extracts. Crude plant extracts made from leaves and roots of rice, respectively, were incubated at 40°C for 30 min as described in . The reaction mixture (1 µl) was resolved on a TLC plate. Purified OsCERK recombinant protein was used as the positive control (+CK). For the negative control (−CK) no extract was added. (F) Comparison of recombinant OsCERK and rACD5. The recombinant protein (1.0 µg) was added in 100 µl reaction system, and incubated at 40°C for 30 min. –CK, no recombinant protein was added. Arrows in (E and F) indicated the formed product <t>C6-NBD-ceramide-1-phosphate</t> (C6-NBD-Cer-1P).
Nbd C6 Ceramide Complex, supplied by Biotium, 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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nbd c6 glccer  (Avanti Polar)
92
Avanti Polar nbd c6 glccer
(A) Schematic representation of the sequence organization of OsCERK . The sizes of the 5′-UTR (white), exons (black) and 3′-UTR (grey) are indicated by numbers below the boxes. The sizes of introns are shown above. (B) Phylogenetic analysis of CERKs and the related enzymes sphingosine kinase (SPK)s, and diacylglycerol kinase (DAGK)s. The architecture of each gene was obtained from SMART using the architecture analysis function, and conserved domains (letters a–d) are indicated as follows: a, Pleckstrin homology domain (PH domain; accession number SM00233); b, Diacylglycerol kinase catalytic domain (DAGKc; SMART accession number SM00046); c, segments of low compositional complexity determined by the program; d, diacylgycerol kinase catalytic domain (DAGK_cat; Pfam accession number PF00781). Detailed sequences information is described in . AtCERK is ACD5. (C) CLUSTAL W alignment of CERK from various organisms showing the highly conserved CXXXCXXC motif. (D) RT-PCR expression analysis of OsCERK in leaves, stem and roots of 2-week-old seedlings. β-actin was used as a control. (E) CERK activity of rice extracts. Crude plant extracts made from leaves and roots of rice, respectively, were incubated at 40°C for 30 min as described in . The reaction mixture (1 µl) was resolved on a TLC plate. Purified OsCERK recombinant protein was used as the positive control (+CK). For the negative control (−CK) no extract was added. (F) Comparison of recombinant OsCERK and rACD5. The recombinant protein (1.0 µg) was added in 100 µl reaction system, and incubated at 40°C for 30 min. –CK, no recombinant protein was added. Arrows in (E and F) indicated the formed product <t>C6-NBD-ceramide-1-phosphate</t> (C6-NBD-Cer-1P).
Nbd C6 Glccer, supplied by Avanti Polar, 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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n d m e t h o d s chemicals 1 acyl  (Avanti Polar)
90
Avanti Polar n d m e t h o d s chemicals 1 acyl
(A) Schematic representation of the sequence organization of OsCERK . The sizes of the 5′-UTR (white), exons (black) and 3′-UTR (grey) are indicated by numbers below the boxes. The sizes of introns are shown above. (B) Phylogenetic analysis of CERKs and the related enzymes sphingosine kinase (SPK)s, and diacylglycerol kinase (DAGK)s. The architecture of each gene was obtained from SMART using the architecture analysis function, and conserved domains (letters a–d) are indicated as follows: a, Pleckstrin homology domain (PH domain; accession number SM00233); b, Diacylglycerol kinase catalytic domain (DAGKc; SMART accession number SM00046); c, segments of low compositional complexity determined by the program; d, diacylgycerol kinase catalytic domain (DAGK_cat; Pfam accession number PF00781). Detailed sequences information is described in . AtCERK is ACD5. (C) CLUSTAL W alignment of CERK from various organisms showing the highly conserved CXXXCXXC motif. (D) RT-PCR expression analysis of OsCERK in leaves, stem and roots of 2-week-old seedlings. β-actin was used as a control. (E) CERK activity of rice extracts. Crude plant extracts made from leaves and roots of rice, respectively, were incubated at 40°C for 30 min as described in . The reaction mixture (1 µl) was resolved on a TLC plate. Purified OsCERK recombinant protein was used as the positive control (+CK). For the negative control (−CK) no extract was added. (F) Comparison of recombinant OsCERK and rACD5. The recombinant protein (1.0 µg) was added in 100 µl reaction system, and incubated at 40°C for 30 min. –CK, no recombinant protein was added. Arrows in (E and F) indicated the formed product <t>C6-NBD-ceramide-1-phosphate</t> (C6-NBD-Cer-1P).
N D M E T H O D S Chemicals 1 Acyl, supplied by Avanti Polar, 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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6-[n-(7-nitro-2,1,3benzoxadiazol-4-yl) amino]-hexanoyl ceramide (c6-nbdcer)  (Matreya LLC)
90
Matreya LLC 6-[n-(7-nitro-2,1,3benzoxadiazol-4-yl) amino]-hexanoyl ceramide (c6-nbdcer)
(A) Schematic representation of the sequence organization of OsCERK . The sizes of the 5′-UTR (white), exons (black) and 3′-UTR (grey) are indicated by numbers below the boxes. The sizes of introns are shown above. (B) Phylogenetic analysis of CERKs and the related enzymes sphingosine kinase (SPK)s, and diacylglycerol kinase (DAGK)s. The architecture of each gene was obtained from SMART using the architecture analysis function, and conserved domains (letters a–d) are indicated as follows: a, Pleckstrin homology domain (PH domain; accession number SM00233); b, Diacylglycerol kinase catalytic domain (DAGKc; SMART accession number SM00046); c, segments of low compositional complexity determined by the program; d, diacylgycerol kinase catalytic domain (DAGK_cat; Pfam accession number PF00781). Detailed sequences information is described in . AtCERK is ACD5. (C) CLUSTAL W alignment of CERK from various organisms showing the highly conserved CXXXCXXC motif. (D) RT-PCR expression analysis of OsCERK in leaves, stem and roots of 2-week-old seedlings. β-actin was used as a control. (E) CERK activity of rice extracts. Crude plant extracts made from leaves and roots of rice, respectively, were incubated at 40°C for 30 min as described in . The reaction mixture (1 µl) was resolved on a TLC plate. Purified OsCERK recombinant protein was used as the positive control (+CK). For the negative control (−CK) no extract was added. (F) Comparison of recombinant OsCERK and rACD5. The recombinant protein (1.0 µg) was added in 100 µl reaction system, and incubated at 40°C for 30 min. –CK, no recombinant protein was added. Arrows in (E and F) indicated the formed product <t>C6-NBD-ceramide-1-phosphate</t> (C6-NBD-Cer-1P).
6 [N (7 Nitro 2,1,3benzoxadiazol 4 Yl) Amino] Hexanoyl Ceramide (C6 Nbdcer), supplied by Matreya LLC, 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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c6-nbd-ceramide  (Interchim Chemicals)
90
Interchim Chemicals c6-nbd-ceramide
(A) Schematic representation of the sequence organization of OsCERK . The sizes of the 5′-UTR (white), exons (black) and 3′-UTR (grey) are indicated by numbers below the boxes. The sizes of introns are shown above. (B) Phylogenetic analysis of CERKs and the related enzymes sphingosine kinase (SPK)s, and diacylglycerol kinase (DAGK)s. The architecture of each gene was obtained from SMART using the architecture analysis function, and conserved domains (letters a–d) are indicated as follows: a, Pleckstrin homology domain (PH domain; accession number SM00233); b, Diacylglycerol kinase catalytic domain (DAGKc; SMART accession number SM00046); c, segments of low compositional complexity determined by the program; d, diacylgycerol kinase catalytic domain (DAGK_cat; Pfam accession number PF00781). Detailed sequences information is described in . AtCERK is ACD5. (C) CLUSTAL W alignment of CERK from various organisms showing the highly conserved CXXXCXXC motif. (D) RT-PCR expression analysis of OsCERK in leaves, stem and roots of 2-week-old seedlings. β-actin was used as a control. (E) CERK activity of rice extracts. Crude plant extracts made from leaves and roots of rice, respectively, were incubated at 40°C for 30 min as described in . The reaction mixture (1 µl) was resolved on a TLC plate. Purified OsCERK recombinant protein was used as the positive control (+CK). For the negative control (−CK) no extract was added. (F) Comparison of recombinant OsCERK and rACD5. The recombinant protein (1.0 µg) was added in 100 µl reaction system, and incubated at 40°C for 30 min. –CK, no recombinant protein was added. Arrows in (E and F) indicated the formed product <t>C6-NBD-ceramide-1-phosphate</t> (C6-NBD-Cer-1P).
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Image Search Results


(A) Schematic representation of the sequence organization of OsCERK . The sizes of the 5′-UTR (white), exons (black) and 3′-UTR (grey) are indicated by numbers below the boxes. The sizes of introns are shown above. (B) Phylogenetic analysis of CERKs and the related enzymes sphingosine kinase (SPK)s, and diacylglycerol kinase (DAGK)s. The architecture of each gene was obtained from SMART using the architecture analysis function, and conserved domains (letters a–d) are indicated as follows: a, Pleckstrin homology domain (PH domain; accession number SM00233); b, Diacylglycerol kinase catalytic domain (DAGKc; SMART accession number SM00046); c, segments of low compositional complexity determined by the program; d, diacylgycerol kinase catalytic domain (DAGK_cat; Pfam accession number PF00781). Detailed sequences information is described in . AtCERK is ACD5. (C) CLUSTAL W alignment of CERK from various organisms showing the highly conserved CXXXCXXC motif. (D) RT-PCR expression analysis of OsCERK in leaves, stem and roots of 2-week-old seedlings. β-actin was used as a control. (E) CERK activity of rice extracts. Crude plant extracts made from leaves and roots of rice, respectively, were incubated at 40°C for 30 min as described in . The reaction mixture (1 µl) was resolved on a TLC plate. Purified OsCERK recombinant protein was used as the positive control (+CK). For the negative control (−CK) no extract was added. (F) Comparison of recombinant OsCERK and rACD5. The recombinant protein (1.0 µg) was added in 100 µl reaction system, and incubated at 40°C for 30 min. –CK, no recombinant protein was added. Arrows in (E and F) indicated the formed product C6-NBD-ceramide-1-phosphate (C6-NBD-Cer-1P).

Journal: PLoS ONE

Article Title: A Conserved Cysteine Motif Is Critical for Rice Ceramide Kinase Activity and Function

doi: 10.1371/journal.pone.0018079

Figure Lengend Snippet: (A) Schematic representation of the sequence organization of OsCERK . The sizes of the 5′-UTR (white), exons (black) and 3′-UTR (grey) are indicated by numbers below the boxes. The sizes of introns are shown above. (B) Phylogenetic analysis of CERKs and the related enzymes sphingosine kinase (SPK)s, and diacylglycerol kinase (DAGK)s. The architecture of each gene was obtained from SMART using the architecture analysis function, and conserved domains (letters a–d) are indicated as follows: a, Pleckstrin homology domain (PH domain; accession number SM00233); b, Diacylglycerol kinase catalytic domain (DAGKc; SMART accession number SM00046); c, segments of low compositional complexity determined by the program; d, diacylgycerol kinase catalytic domain (DAGK_cat; Pfam accession number PF00781). Detailed sequences information is described in . AtCERK is ACD5. (C) CLUSTAL W alignment of CERK from various organisms showing the highly conserved CXXXCXXC motif. (D) RT-PCR expression analysis of OsCERK in leaves, stem and roots of 2-week-old seedlings. β-actin was used as a control. (E) CERK activity of rice extracts. Crude plant extracts made from leaves and roots of rice, respectively, were incubated at 40°C for 30 min as described in . The reaction mixture (1 µl) was resolved on a TLC plate. Purified OsCERK recombinant protein was used as the positive control (+CK). For the negative control (−CK) no extract was added. (F) Comparison of recombinant OsCERK and rACD5. The recombinant protein (1.0 µg) was added in 100 µl reaction system, and incubated at 40°C for 30 min. –CK, no recombinant protein was added. Arrows in (E and F) indicated the formed product C6-NBD-ceramide-1-phosphate (C6-NBD-Cer-1P).

Article Snippet: The C6-NBD-ceramide-1-phosphate products were identified by comparison with C6-NBD-ceramide (Avanti Polar lipids, Alabaster, AL, USA) and quantified using IQTL software (GE Healthcare).

Techniques: Sequencing, Reverse Transcription Polymerase Chain Reaction, Expressing, Activity Assay, Incubation, Purification, Recombinant, Positive Control, Negative Control

(A) Effects of cycloheximide (CHX) on C6 ceramide-induced cell death. (B) Effects of K252a on C6 ceramide-induced cell death. (C) Effects of C2 ceramide-1-phosphate on C2 ceramide induced cell death. Protoplasts were treated with less than 0.15% solvent or the indicated reagent. Percentage of surviving cells was estimated by FDA staining. These assays were repeated three times with similar results. Letters indicate that values are different using Fisher's PLSD test ( P <0.05). Bars show standard deviations.

Journal: PLoS ONE

Article Title: A Conserved Cysteine Motif Is Critical for Rice Ceramide Kinase Activity and Function

doi: 10.1371/journal.pone.0018079

Figure Lengend Snippet: (A) Effects of cycloheximide (CHX) on C6 ceramide-induced cell death. (B) Effects of K252a on C6 ceramide-induced cell death. (C) Effects of C2 ceramide-1-phosphate on C2 ceramide induced cell death. Protoplasts were treated with less than 0.15% solvent or the indicated reagent. Percentage of surviving cells was estimated by FDA staining. These assays were repeated three times with similar results. Letters indicate that values are different using Fisher's PLSD test ( P <0.05). Bars show standard deviations.

Article Snippet: The C6-NBD-ceramide-1-phosphate products were identified by comparison with C6-NBD-ceramide (Avanti Polar lipids, Alabaster, AL, USA) and quantified using IQTL software (GE Healthcare).

Techniques: Staining