anti-caga antibodies Search Results


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    Helicobacter pylori CagA Protein mouse monoclonal antibody clone 10E9 Aff Purified
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    88
    Austral Biologicals anti caga antibody
    Translocation of <t>CagA</t> into AGS cells. AGS gastric cells were cocultured with a wild-type H. pylori strain ( H. pylori strain 26695 Sc#7) (WT), with an isogenic mutant strain that lacked the cag PAI (Δ cag PAI), or without bacteria. After 6 h, the cells were lysed, and the proteins in the lysates were analyzed by immunoblotting with an anti-phosphotyrosine (anti-PY99) antibody (top panel). The membrane was then stripped and <t>immunoblotted</t> with an anti-CagA antibody (bottom panel). The wild-type strain expressed a CagA protein that underwent tyrosine phosphorylation (CagA p-Tyr ), and the isogenic cag PAI deletion mutant strain did not express CagA.
    Anti Caga Antibody, supplied by Austral Biologicals, used in various techniques. Bioz Stars score: 88/100, based on 53 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Austral Biologicals rabbit polyclonal α caga antibody
    Elevated E-cadherin cleavage and <t>CagA</t> translocation in host cells by overexpression of HtrA. Polarized MKN-28 cells were infected with IPTG-induced or control Hp wt or Hp htrA 26695 for 24 h. a Cell pellets and supernatants were prepared and subjected to α-E-cadherin Western blotting. The full-length E-cadherin signals (130 kDa) in the cell pellets and cleaved-off NTF-domain (90 kDa) in the culture supernatants are marked with arrows . b Translocation and phosphorylation of CagA ( arrow ) were analyzed using α-PY-99 and <t>α-CagA</t> antibodies, respectively. The asterisk marks a phosphorylated 125 kDa host cell protein migrating below CagA. c The band intensities of cleaved E-cadherin NTF-fragments and phospho-CagA signals were quantified densitometrically. The relative amounts of NTF-fragment and phosphorylated CagA are given in “fold change”. All experiments were done in triplicates
    Rabbit Polyclonal α Caga Antibody, supplied by Austral Biologicals, used in various techniques. Bioz Stars score: 88/100, based on 38 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    85
    Austral Biologicals anti caga polyclonal antibodies
    <t>CagA</t> with two variable-region TPMs is less phosphorylated than CagA with three variable-region TPMs. (A) AGS cells were cocultured with H. pylori strains possessing two (strain Z11) or three (strains Z4, Z7, and Z23) variable-region EPIYA motifs for 6 h at 37°C before the cells were lysed and the samples were subjected to sodium dodecyl sulfate-polyacrylamide gel electrophoresis and Western blotting with antiphosphotyrosine monoclonal antibodies (lower panel). The blots were then stripped and reprobed with anti-CagA <t>polyclonal</t> antibodies (upper panel). (B) Densitometric analysis of the degree of CagA phosphorylation, expressed as a ratio of phospho-Tyr intensity to CagA protein intensity. The data shown are representative of those from three separate experiments.
    Anti Caga Polyclonal Antibodies, supplied by Austral Biologicals, used in various techniques. Bioz Stars score: 85/100, based on 7 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    88
    Austral Biologicals pylori caga antibody
    <t>CagA</t> with two variable-region TPMs is less phosphorylated than CagA with three variable-region TPMs. (A) AGS cells were cocultured with H. pylori strains possessing two (strain Z11) or three (strains Z4, Z7, and Z23) variable-region EPIYA motifs for 6 h at 37°C before the cells were lysed and the samples were subjected to sodium dodecyl sulfate-polyacrylamide gel electrophoresis and Western blotting with antiphosphotyrosine monoclonal antibodies (lower panel). The blots were then stripped and reprobed with anti-CagA <t>polyclonal</t> antibodies (upper panel). (B) Densitometric analysis of the degree of CagA phosphorylation, expressed as a ratio of phospho-Tyr intensity to CagA protein intensity. The data shown are representative of those from three separate experiments.
    Pylori Caga Antibody, supplied by Austral Biologicals, used in various techniques. Bioz Stars score: 88/100, based on 31 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    92
    Abcam anti caga antibody
    <t>CagA</t> with two variable-region TPMs is less phosphorylated than CagA with three variable-region TPMs. (A) AGS cells were cocultured with H. pylori strains possessing two (strain Z11) or three (strains Z4, Z7, and Z23) variable-region EPIYA motifs for 6 h at 37°C before the cells were lysed and the samples were subjected to sodium dodecyl sulfate-polyacrylamide gel electrophoresis and Western blotting with antiphosphotyrosine monoclonal antibodies (lower panel). The blots were then stripped and reprobed with anti-CagA <t>polyclonal</t> antibodies (upper panel). (B) Densitometric analysis of the degree of CagA phosphorylation, expressed as a ratio of phospho-Tyr intensity to CagA protein intensity. The data shown are representative of those from three separate experiments.
    Anti Caga Antibody, supplied by Abcam, used in various techniques. Bioz Stars score: 92/100, based on 4 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Abcam monoclonal anti caga antibody
    <t>CagA</t> with two variable-region TPMs is less phosphorylated than CagA with three variable-region TPMs. (A) AGS cells were cocultured with H. pylori strains possessing two (strain Z11) or three (strains Z4, Z7, and Z23) variable-region EPIYA motifs for 6 h at 37°C before the cells were lysed and the samples were subjected to sodium dodecyl sulfate-polyacrylamide gel electrophoresis and Western blotting with antiphosphotyrosine monoclonal antibodies (lower panel). The blots were then stripped and reprobed with anti-CagA <t>polyclonal</t> antibodies (upper panel). (B) Densitometric analysis of the degree of CagA phosphorylation, expressed as a ratio of phospho-Tyr intensity to CagA protein intensity. The data shown are representative of those from three separate experiments.
    Monoclonal Anti Caga Antibody, supplied by Abcam, used in various techniques. Bioz Stars score: 86/100, based on 13 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Austral Biologicals monoclonal anti caga antibody
    Generation of phosphomimetic <t>CagA</t> mutants and their interaction with host signaling factors during H. pylori infection. ( A ) Site-directed Y > D mutagenesis of CagA EPIYA-A, EPIYA-B, and EPIYA-C to generate phosphomimetic mutants. Nontargeted tyrosines in adjacent EPIYA motifs were replaced by phenylalanines to avoid additional phosphorylation events per molecule. The resulting single, double, and triple mutants were named as indicated. ( B ) AGS cells were infected for 4 hours with CagA-expressing H. pylori strains as indicated. CagA phosphorylation was examined using <t>α–PY-99</t> and α-CagA antibodies (arrows). All strains expressed similar amounts of CagA, but only H. pylori expressing WT CagA revealed a phosphorylation signal. The asterisk in the lower panel indicates antibody cross-reactivity with an unknown phosphorylated host cell protein. ( C – E ) AGS cells were infected for 4 hours with CagA-expressing H. pylori strains as indicated, and cell lysates were subjected to IP with α-CagA antibodies. Western blotting using α-Csk ( C ), α-PI3K ( D ), or α–SHP-2 ( E ) indicated that each of these factors formed a complex with the respective phosphomimetic CagA EPIYA Y > D mutant, but not with the nonphosphorylatable triple EPIYA-ABC Y > F mutant as control.
    Monoclonal Anti Caga Antibody, supplied by Austral Biologicals, used in various techniques. Bioz Stars score: 85/100, based on 16 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Abcam mouse monoclonal antibody against human caga
    Generation of phosphomimetic <t>CagA</t> mutants and their interaction with host signaling factors during H. pylori infection. ( A ) Site-directed Y > D mutagenesis of CagA EPIYA-A, EPIYA-B, and EPIYA-C to generate phosphomimetic mutants. Nontargeted tyrosines in adjacent EPIYA motifs were replaced by phenylalanines to avoid additional phosphorylation events per molecule. The resulting single, double, and triple mutants were named as indicated. ( B ) AGS cells were infected for 4 hours with CagA-expressing H. pylori strains as indicated. CagA phosphorylation was examined using <t>α–PY-99</t> and α-CagA antibodies (arrows). All strains expressed similar amounts of CagA, but only H. pylori expressing WT CagA revealed a phosphorylation signal. The asterisk in the lower panel indicates antibody cross-reactivity with an unknown phosphorylated host cell protein. ( C – E ) AGS cells were infected for 4 hours with CagA-expressing H. pylori strains as indicated, and cell lysates were subjected to IP with α-CagA antibodies. Western blotting using α-Csk ( C ), α-PI3K ( D ), or α–SHP-2 ( E ) indicated that each of these factors formed a complex with the respective phosphomimetic CagA EPIYA Y > D mutant, but not with the nonphosphorylatable triple EPIYA-ABC Y > F mutant as control.
    Mouse Monoclonal Antibody Against Human Caga, supplied by Abcam, used in various techniques. Bioz Stars score: 99/100, based on 8 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Austral Biologicals mouse monoclonal α caga antibodies
    Sequence comparison of the three TPM sites in <t>CagA</t> proteins from different clinical H. pylori strains and specific detection of phosphorylated EPIYT-motif during co-culture. Panel A : CagA proteins of H. pylori vary in their carboxy-terminal TPM sites. These EPIYA-repeats serve as tyrosine phosphorylation sites of CagA and can be targeted by c-Abl and c-Src kinases. Three EPIYA- or EPIYT-segments at position A, B and C are shaded with yellow. One striking feature of B-TPM is the presence of a threonine residue in the +1 position (shaded with blue) relative to the phosphorylated tyrosine residue, which is highly conserved in most but not all H. pylori strains and may affect the capabilities of binding the p85 subunit of PI3-kinase, as discussed in the text. The CagA protein sequences were obtained from databases and sequence alignment was done using the ClustalW2 program ( http://www.ebi.ac.uk/Tools/msa/clustalw2/ ). Panel B : To investigate whether the EPIYT-motif can be phosphorylated during co-culture, AGS cells were co-incubated with the indicated CagA-expressing H. pylori strains for 6 h. Phosphorylation of CagA was examined using the phospho-specific α-pCagA-EPIYT-918 antibody. Loading of equal amounts of protein in each sample was confirmed by probing with monoclonal <t>α-CagA</t> and α-GAPDH antibodies.
    Mouse Monoclonal α Caga Antibodies, supplied by Austral Biologicals, used in various techniques. Bioz Stars score: 88/100, based on 44 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    92
    Abnova pylori caga antibody
    Sequence comparison of the three TPM sites in <t>CagA</t> proteins from different clinical H. pylori strains and specific detection of phosphorylated EPIYT-motif during co-culture. Panel A : CagA proteins of H. pylori vary in their carboxy-terminal TPM sites. These EPIYA-repeats serve as tyrosine phosphorylation sites of CagA and can be targeted by c-Abl and c-Src kinases. Three EPIYA- or EPIYT-segments at position A, B and C are shaded with yellow. One striking feature of B-TPM is the presence of a threonine residue in the +1 position (shaded with blue) relative to the phosphorylated tyrosine residue, which is highly conserved in most but not all H. pylori strains and may affect the capabilities of binding the p85 subunit of PI3-kinase, as discussed in the text. The CagA protein sequences were obtained from databases and sequence alignment was done using the ClustalW2 program ( http://www.ebi.ac.uk/Tools/msa/clustalw2/ ). Panel B : To investigate whether the EPIYT-motif can be phosphorylated during co-culture, AGS cells were co-incubated with the indicated CagA-expressing H. pylori strains for 6 h. Phosphorylation of CagA was examined using the phospho-specific α-pCagA-EPIYT-918 antibody. Loading of equal amounts of protein in each sample was confirmed by probing with monoclonal <t>α-CagA</t> and α-GAPDH antibodies.
    Pylori Caga Antibody, supplied by Abnova, used in various techniques. Bioz Stars score: 92/100, based on 12 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    80
    Oravax Inc anti recombinant caga protein mouse polyclonal antibody
    Sequence comparison of the three TPM sites in <t>CagA</t> proteins from different clinical H. pylori strains and specific detection of phosphorylated EPIYT-motif during co-culture. Panel A : CagA proteins of H. pylori vary in their carboxy-terminal TPM sites. These EPIYA-repeats serve as tyrosine phosphorylation sites of CagA and can be targeted by c-Abl and c-Src kinases. Three EPIYA- or EPIYT-segments at position A, B and C are shaded with yellow. One striking feature of B-TPM is the presence of a threonine residue in the +1 position (shaded with blue) relative to the phosphorylated tyrosine residue, which is highly conserved in most but not all H. pylori strains and may affect the capabilities of binding the p85 subunit of PI3-kinase, as discussed in the text. The CagA protein sequences were obtained from databases and sequence alignment was done using the ClustalW2 program ( http://www.ebi.ac.uk/Tools/msa/clustalw2/ ). Panel B : To investigate whether the EPIYT-motif can be phosphorylated during co-culture, AGS cells were co-incubated with the indicated CagA-expressing H. pylori strains for 6 h. Phosphorylation of CagA was examined using the phospho-specific α-pCagA-EPIYT-918 antibody. Loading of equal amounts of protein in each sample was confirmed by probing with monoclonal <t>α-CagA</t> and α-GAPDH antibodies.
    Anti Recombinant Caga Protein Mouse Polyclonal Antibody, supplied by Oravax Inc, used in various techniques. Bioz Stars score: 80/100, based on 4 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    88
    Santa Cruz Biotechnology anti caga antibody
    Sequence comparison of the three TPM sites in <t>CagA</t> proteins from different clinical H. pylori strains and specific detection of phosphorylated EPIYT-motif during co-culture. Panel A : CagA proteins of H. pylori vary in their carboxy-terminal TPM sites. These EPIYA-repeats serve as tyrosine phosphorylation sites of CagA and can be targeted by c-Abl and c-Src kinases. Three EPIYA- or EPIYT-segments at position A, B and C are shaded with yellow. One striking feature of B-TPM is the presence of a threonine residue in the +1 position (shaded with blue) relative to the phosphorylated tyrosine residue, which is highly conserved in most but not all H. pylori strains and may affect the capabilities of binding the p85 subunit of PI3-kinase, as discussed in the text. The CagA protein sequences were obtained from databases and sequence alignment was done using the ClustalW2 program ( http://www.ebi.ac.uk/Tools/msa/clustalw2/ ). Panel B : To investigate whether the EPIYT-motif can be phosphorylated during co-culture, AGS cells were co-incubated with the indicated CagA-expressing H. pylori strains for 6 h. Phosphorylation of CagA was examined using the phospho-specific α-pCagA-EPIYT-918 antibody. Loading of equal amounts of protein in each sample was confirmed by probing with monoclonal <t>α-CagA</t> and α-GAPDH antibodies.
    Anti Caga Antibody, supplied by Santa Cruz Biotechnology, used in various techniques. Bioz Stars score: 88/100, based on 67 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Austral Biologicals anti caga polyclonal antibody hpp 5003 9
    Sequence comparison of the three TPM sites in <t>CagA</t> proteins from different clinical H. pylori strains and specific detection of phosphorylated EPIYT-motif during co-culture. Panel A : CagA proteins of H. pylori vary in their carboxy-terminal TPM sites. These EPIYA-repeats serve as tyrosine phosphorylation sites of CagA and can be targeted by c-Abl and c-Src kinases. Three EPIYA- or EPIYT-segments at position A, B and C are shaded with yellow. One striking feature of B-TPM is the presence of a threonine residue in the +1 position (shaded with blue) relative to the phosphorylated tyrosine residue, which is highly conserved in most but not all H. pylori strains and may affect the capabilities of binding the p85 subunit of PI3-kinase, as discussed in the text. The CagA protein sequences were obtained from databases and sequence alignment was done using the ClustalW2 program ( http://www.ebi.ac.uk/Tools/msa/clustalw2/ ). Panel B : To investigate whether the EPIYT-motif can be phosphorylated during co-culture, AGS cells were co-incubated with the indicated CagA-expressing H. pylori strains for 6 h. Phosphorylation of CagA was examined using the phospho-specific α-pCagA-EPIYT-918 antibody. Loading of equal amounts of protein in each sample was confirmed by probing with monoclonal <t>α-CagA</t> and α-GAPDH antibodies.
    Anti Caga Polyclonal Antibody Hpp 5003 9, supplied by Austral Biologicals, used in various techniques. Bioz Stars score: 85/100, based on 2 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    85
    Novartis anti caga rabbit polyclonal antibodies
    Sequence comparison of the three TPM sites in <t>CagA</t> proteins from different clinical H. pylori strains and specific detection of phosphorylated EPIYT-motif during co-culture. Panel A : CagA proteins of H. pylori vary in their carboxy-terminal TPM sites. These EPIYA-repeats serve as tyrosine phosphorylation sites of CagA and can be targeted by c-Abl and c-Src kinases. Three EPIYA- or EPIYT-segments at position A, B and C are shaded with yellow. One striking feature of B-TPM is the presence of a threonine residue in the +1 position (shaded with blue) relative to the phosphorylated tyrosine residue, which is highly conserved in most but not all H. pylori strains and may affect the capabilities of binding the p85 subunit of PI3-kinase, as discussed in the text. The CagA protein sequences were obtained from databases and sequence alignment was done using the ClustalW2 program ( http://www.ebi.ac.uk/Tools/msa/clustalw2/ ). Panel B : To investigate whether the EPIYT-motif can be phosphorylated during co-culture, AGS cells were co-incubated with the indicated CagA-expressing H. pylori strains for 6 h. Phosphorylation of CagA was examined using the phospho-specific α-pCagA-EPIYT-918 antibody. Loading of equal amounts of protein in each sample was confirmed by probing with monoclonal <t>α-CagA</t> and α-GAPDH antibodies.
    Anti Caga Rabbit Polyclonal Antibodies, supplied by Novartis, used in various techniques. Bioz Stars score: 85/100, based on 2 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    90
    Agilent technologies anti east asian type caga specific antibody
    Sequence comparison of the three TPM sites in <t>CagA</t> proteins from different clinical H. pylori strains and specific detection of phosphorylated EPIYT-motif during co-culture. Panel A : CagA proteins of H. pylori vary in their carboxy-terminal TPM sites. These EPIYA-repeats serve as tyrosine phosphorylation sites of CagA and can be targeted by c-Abl and c-Src kinases. Three EPIYA- or EPIYT-segments at position A, B and C are shaded with yellow. One striking feature of B-TPM is the presence of a threonine residue in the +1 position (shaded with blue) relative to the phosphorylated tyrosine residue, which is highly conserved in most but not all H. pylori strains and may affect the capabilities of binding the p85 subunit of PI3-kinase, as discussed in the text. The CagA protein sequences were obtained from databases and sequence alignment was done using the ClustalW2 program ( http://www.ebi.ac.uk/Tools/msa/clustalw2/ ). Panel B : To investigate whether the EPIYT-motif can be phosphorylated during co-culture, AGS cells were co-incubated with the indicated CagA-expressing H. pylori strains for 6 h. Phosphorylation of CagA was examined using the phospho-specific α-pCagA-EPIYT-918 antibody. Loading of equal amounts of protein in each sample was confirmed by probing with monoclonal <t>α-CagA</t> and α-GAPDH antibodies.
    Anti East Asian Type Caga Specific Antibody, supplied by Agilent technologies, used in various techniques. Bioz Stars score: 90/100, based on 8 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Aalto Bio Reagents monoclonal caga antibody
    Sequence comparison of the three TPM sites in <t>CagA</t> proteins from different clinical H. pylori strains and specific detection of phosphorylated EPIYT-motif during co-culture. Panel A : CagA proteins of H. pylori vary in their carboxy-terminal TPM sites. These EPIYA-repeats serve as tyrosine phosphorylation sites of CagA and can be targeted by c-Abl and c-Src kinases. Three EPIYA- or EPIYT-segments at position A, B and C are shaded with yellow. One striking feature of B-TPM is the presence of a threonine residue in the +1 position (shaded with blue) relative to the phosphorylated tyrosine residue, which is highly conserved in most but not all H. pylori strains and may affect the capabilities of binding the p85 subunit of PI3-kinase, as discussed in the text. The CagA protein sequences were obtained from databases and sequence alignment was done using the ClustalW2 program ( http://www.ebi.ac.uk/Tools/msa/clustalw2/ ). Panel B : To investigate whether the EPIYT-motif can be phosphorylated during co-culture, AGS cells were co-incubated with the indicated CagA-expressing H. pylori strains for 6 h. Phosphorylation of CagA was examined using the phospho-specific α-pCagA-EPIYT-918 antibody. Loading of equal amounts of protein in each sample was confirmed by probing with monoclonal <t>α-CagA</t> and α-GAPDH antibodies.
    Monoclonal Caga Antibody, supplied by Aalto Bio Reagents, used in various techniques. Bioz Stars score: 85/100, based on 5 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Dia Pro Diagnostic Bioprobes Srl igg antibodies against caga protein
    Sequence comparison of the three TPM sites in <t>CagA</t> proteins from different clinical H. pylori strains and specific detection of phosphorylated EPIYT-motif during co-culture. Panel A : CagA proteins of H. pylori vary in their carboxy-terminal TPM sites. These EPIYA-repeats serve as tyrosine phosphorylation sites of CagA and can be targeted by c-Abl and c-Src kinases. Three EPIYA- or EPIYT-segments at position A, B and C are shaded with yellow. One striking feature of B-TPM is the presence of a threonine residue in the +1 position (shaded with blue) relative to the phosphorylated tyrosine residue, which is highly conserved in most but not all H. pylori strains and may affect the capabilities of binding the p85 subunit of PI3-kinase, as discussed in the text. The CagA protein sequences were obtained from databases and sequence alignment was done using the ClustalW2 program ( http://www.ebi.ac.uk/Tools/msa/clustalw2/ ). Panel B : To investigate whether the EPIYT-motif can be phosphorylated during co-culture, AGS cells were co-incubated with the indicated CagA-expressing H. pylori strains for 6 h. Phosphorylation of CagA was examined using the phospho-specific α-pCagA-EPIYT-918 antibody. Loading of equal amounts of protein in each sample was confirmed by probing with monoclonal <t>α-CagA</t> and α-GAPDH antibodies.
    Igg Antibodies Against Caga Protein, supplied by Dia Pro Diagnostic Bioprobes Srl, used in various techniques. Bioz Stars score: 85/100, based on 6 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    85
    Austral Biologicals rabbit anti caga antibody
    Sequence comparison of the three TPM sites in <t>CagA</t> proteins from different clinical H. pylori strains and specific detection of phosphorylated EPIYT-motif during co-culture. Panel A : CagA proteins of H. pylori vary in their carboxy-terminal TPM sites. These EPIYA-repeats serve as tyrosine phosphorylation sites of CagA and can be targeted by c-Abl and c-Src kinases. Three EPIYA- or EPIYT-segments at position A, B and C are shaded with yellow. One striking feature of B-TPM is the presence of a threonine residue in the +1 position (shaded with blue) relative to the phosphorylated tyrosine residue, which is highly conserved in most but not all H. pylori strains and may affect the capabilities of binding the p85 subunit of PI3-kinase, as discussed in the text. The CagA protein sequences were obtained from databases and sequence alignment was done using the ClustalW2 program ( http://www.ebi.ac.uk/Tools/msa/clustalw2/ ). Panel B : To investigate whether the EPIYT-motif can be phosphorylated during co-culture, AGS cells were co-incubated with the indicated CagA-expressing H. pylori strains for 6 h. Phosphorylation of CagA was examined using the phospho-specific α-pCagA-EPIYT-918 antibody. Loading of equal amounts of protein in each sample was confirmed by probing with monoclonal <t>α-CagA</t> and α-GAPDH antibodies.
    Rabbit Anti Caga Antibody, supplied by Austral Biologicals, used in various techniques. Bioz Stars score: 85/100, based on 6 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/rabbit anti caga antibody/product/Austral Biologicals
    Average 85 stars, based on 6 article reviews
    Price from $9.99 to $1999.99
    rabbit anti caga antibody - by Bioz Stars, 2021-01
    85/100 stars
      Buy from Supplier

    85
    Austral Biologicals rabbit anti caga peptide antibody
    Sequence comparison of the three TPM sites in <t>CagA</t> proteins from different clinical H. pylori strains and specific detection of phosphorylated EPIYT-motif during co-culture. Panel A : CagA proteins of H. pylori vary in their carboxy-terminal TPM sites. These EPIYA-repeats serve as tyrosine phosphorylation sites of CagA and can be targeted by c-Abl and c-Src kinases. Three EPIYA- or EPIYT-segments at position A, B and C are shaded with yellow. One striking feature of B-TPM is the presence of a threonine residue in the +1 position (shaded with blue) relative to the phosphorylated tyrosine residue, which is highly conserved in most but not all H. pylori strains and may affect the capabilities of binding the p85 subunit of PI3-kinase, as discussed in the text. The CagA protein sequences were obtained from databases and sequence alignment was done using the ClustalW2 program ( http://www.ebi.ac.uk/Tools/msa/clustalw2/ ). Panel B : To investigate whether the EPIYT-motif can be phosphorylated during co-culture, AGS cells were co-incubated with the indicated CagA-expressing H. pylori strains for 6 h. Phosphorylation of CagA was examined using the phospho-specific α-pCagA-EPIYT-918 antibody. Loading of equal amounts of protein in each sample was confirmed by probing with monoclonal <t>α-CagA</t> and α-GAPDH antibodies.
    Rabbit Anti Caga Peptide Antibody, supplied by Austral Biologicals, used in various techniques. Bioz Stars score: 85/100, based on 4 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/rabbit anti caga peptide antibody/product/Austral Biologicals
    Average 85 stars, based on 4 article reviews
    Price from $9.99 to $1999.99
    rabbit anti caga peptide antibody - by Bioz Stars, 2021-01
    85/100 stars
      Buy from Supplier

    N/A
    The protein encoded by this gene is a member of the S100 family of proteins containing 2 EF hand calcium binding motifs S100 proteins are localized in the cytoplasm and
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    Translocation of CagA into AGS cells. AGS gastric cells were cocultured with a wild-type H. pylori strain ( H. pylori strain 26695 Sc#7) (WT), with an isogenic mutant strain that lacked the cag PAI (Δ cag PAI), or without bacteria. After 6 h, the cells were lysed, and the proteins in the lysates were analyzed by immunoblotting with an anti-phosphotyrosine (anti-PY99) antibody (top panel). The membrane was then stripped and immunoblotted with an anti-CagA antibody (bottom panel). The wild-type strain expressed a CagA protein that underwent tyrosine phosphorylation (CagA p-Tyr ), and the isogenic cag PAI deletion mutant strain did not express CagA.

    Journal: Journal of Bacteriology

    Article Title: Protein-Protein Interactions among Helicobacter pylori Cag Proteins

    doi: 10.1128/JB.00066-06

    Figure Lengend Snippet: Translocation of CagA into AGS cells. AGS gastric cells were cocultured with a wild-type H. pylori strain ( H. pylori strain 26695 Sc#7) (WT), with an isogenic mutant strain that lacked the cag PAI (Δ cag PAI), or without bacteria. After 6 h, the cells were lysed, and the proteins in the lysates were analyzed by immunoblotting with an anti-phosphotyrosine (anti-PY99) antibody (top panel). The membrane was then stripped and immunoblotted with an anti-CagA antibody (bottom panel). The wild-type strain expressed a CagA protein that underwent tyrosine phosphorylation (CagA p-Tyr ), and the isogenic cag PAI deletion mutant strain did not express CagA.

    Article Snippet: After incubation of the membrane with a stripping buffer, the membrane was immunoblotted with an anti-CagA antibody (Austral Biologicals), followed by horseradish peroxidase-conjugated goat anti-rabbit IgG (Bio-Rad), each used at a dilution of 1:6,000 in TBS-Tween containing 2% nonfat powdered milk.

    Techniques: Translocation Assay, Mutagenesis

    Elevated E-cadherin cleavage and CagA translocation in host cells by overexpression of HtrA. Polarized MKN-28 cells were infected with IPTG-induced or control Hp wt or Hp htrA 26695 for 24 h. a Cell pellets and supernatants were prepared and subjected to α-E-cadherin Western blotting. The full-length E-cadherin signals (130 kDa) in the cell pellets and cleaved-off NTF-domain (90 kDa) in the culture supernatants are marked with arrows . b Translocation and phosphorylation of CagA ( arrow ) were analyzed using α-PY-99 and α-CagA antibodies, respectively. The asterisk marks a phosphorylated 125 kDa host cell protein migrating below CagA. c The band intensities of cleaved E-cadherin NTF-fragments and phospho-CagA signals were quantified densitometrically. The relative amounts of NTF-fragment and phosphorylated CagA are given in “fold change”. All experiments were done in triplicates

    Journal: Gut Pathogens

    Article Title: Overexpression of serine protease HtrA enhances disruption of adherens junctions, paracellular transmigration and type IV secretion of CagA by Helicobacter pylori

    doi: 10.1186/s13099-017-0189-6

    Figure Lengend Snippet: Elevated E-cadherin cleavage and CagA translocation in host cells by overexpression of HtrA. Polarized MKN-28 cells were infected with IPTG-induced or control Hp wt or Hp htrA 26695 for 24 h. a Cell pellets and supernatants were prepared and subjected to α-E-cadherin Western blotting. The full-length E-cadherin signals (130 kDa) in the cell pellets and cleaved-off NTF-domain (90 kDa) in the culture supernatants are marked with arrows . b Translocation and phosphorylation of CagA ( arrow ) were analyzed using α-PY-99 and α-CagA antibodies, respectively. The asterisk marks a phosphorylated 125 kDa host cell protein migrating below CagA. c The band intensities of cleaved E-cadherin NTF-fragments and phospho-CagA signals were quantified densitometrically. The relative amounts of NTF-fragment and phosphorylated CagA are given in “fold change”. All experiments were done in triplicates

    Article Snippet: Antibodies The following antibodies were purchased: rabbit polyclonal α-CagA antibody (Austral Biologicals, San Ramon, CA/USA), monoclonal pan-phosphotyrosine α-PY99 (Santa Cruz, Santa Cruz, CA/USA), rabbit α-GAPDH (Santa Cruz), rabbit α-H. pylori (Dako, Glostrup/Denmark) and two monoclonal antibodies directed against the extracellular domain of E-cadherin, H-108 (Santa Cruz) and CD324 (BD Biosciences, San Jose, CA/USA).

    Techniques: Translocation Assay, Over Expression, Infection, Western Blot

    Chromosomal introduction of a second, inducible htrA gene copy in H. pylori. a Schematic presentation of the IPTG-inducible htrA 26695 expression construct with the chloramphenicol acetyl-transferase (CAT) cassette ( top ), which was cloned into the plasticity region of H. pylori between genes HP0999 and HP1000 ( bottom ). b This construct was transformed into H. pylori wild-type ( Hp wt) strains 26695 and P12, and the expression of HtrA was investigated after 24 h growth in BHI medium in the presence or absence of 1 or 2 mM IPTG, respectively. The results for strain 26695 are shown. Western blotting using α-FlaA and α-CagA antibodies served as controls, indicating that equal amounts of proteins are present in the samples. c The band intensities of HtrA proteins were quantified densitometrically. The relative protein expression is given in “fold change”

    Journal: Gut Pathogens

    Article Title: Overexpression of serine protease HtrA enhances disruption of adherens junctions, paracellular transmigration and type IV secretion of CagA by Helicobacter pylori

    doi: 10.1186/s13099-017-0189-6

    Figure Lengend Snippet: Chromosomal introduction of a second, inducible htrA gene copy in H. pylori. a Schematic presentation of the IPTG-inducible htrA 26695 expression construct with the chloramphenicol acetyl-transferase (CAT) cassette ( top ), which was cloned into the plasticity region of H. pylori between genes HP0999 and HP1000 ( bottom ). b This construct was transformed into H. pylori wild-type ( Hp wt) strains 26695 and P12, and the expression of HtrA was investigated after 24 h growth in BHI medium in the presence or absence of 1 or 2 mM IPTG, respectively. The results for strain 26695 are shown. Western blotting using α-FlaA and α-CagA antibodies served as controls, indicating that equal amounts of proteins are present in the samples. c The band intensities of HtrA proteins were quantified densitometrically. The relative protein expression is given in “fold change”

    Article Snippet: Antibodies The following antibodies were purchased: rabbit polyclonal α-CagA antibody (Austral Biologicals, San Ramon, CA/USA), monoclonal pan-phosphotyrosine α-PY99 (Santa Cruz, Santa Cruz, CA/USA), rabbit α-GAPDH (Santa Cruz), rabbit α-H. pylori (Dako, Glostrup/Denmark) and two monoclonal antibodies directed against the extracellular domain of E-cadherin, H-108 (Santa Cruz) and CD324 (BD Biosciences, San Jose, CA/USA).

    Techniques: Expressing, Construct, Chloramphenicol Acetyltransferase Assay, Clone Assay, Transformation Assay, Western Blot

    Elevated HtrA secretion levels in H. pylori does not affect the extracellular delivery of VacA and GGT. a H. pylori wild-type strain 26695 ( Hp wt) and 26695 transformed with htrA 26695 were grown for 24 h in BHI broth medium with 1% β-cyclodextrin in the presence or absence of 1 or 2 mM IPTG, respectively. Bacteria-free supernatants were prepared and subjected to Western blotting using α-HtrA, α-VacA and α-GGT antibodies. The blots against α-CagA served as control. The band intensities of secreted HtrA ( b ) as well as VacA and GGT ( c ) were quantified densitometrically. The relative protein expression is given in “fold change”. All secretion assays were done in triplicates

    Journal: Gut Pathogens

    Article Title: Overexpression of serine protease HtrA enhances disruption of adherens junctions, paracellular transmigration and type IV secretion of CagA by Helicobacter pylori

    doi: 10.1186/s13099-017-0189-6

    Figure Lengend Snippet: Elevated HtrA secretion levels in H. pylori does not affect the extracellular delivery of VacA and GGT. a H. pylori wild-type strain 26695 ( Hp wt) and 26695 transformed with htrA 26695 were grown for 24 h in BHI broth medium with 1% β-cyclodextrin in the presence or absence of 1 or 2 mM IPTG, respectively. Bacteria-free supernatants were prepared and subjected to Western blotting using α-HtrA, α-VacA and α-GGT antibodies. The blots against α-CagA served as control. The band intensities of secreted HtrA ( b ) as well as VacA and GGT ( c ) were quantified densitometrically. The relative protein expression is given in “fold change”. All secretion assays were done in triplicates

    Article Snippet: Antibodies The following antibodies were purchased: rabbit polyclonal α-CagA antibody (Austral Biologicals, San Ramon, CA/USA), monoclonal pan-phosphotyrosine α-PY99 (Santa Cruz, Santa Cruz, CA/USA), rabbit α-GAPDH (Santa Cruz), rabbit α-H. pylori (Dako, Glostrup/Denmark) and two monoclonal antibodies directed against the extracellular domain of E-cadherin, H-108 (Santa Cruz) and CD324 (BD Biosciences, San Jose, CA/USA).

    Techniques: Transformation Assay, Western Blot, Expressing

    CagA with two variable-region TPMs is less phosphorylated than CagA with three variable-region TPMs. (A) AGS cells were cocultured with H. pylori strains possessing two (strain Z11) or three (strains Z4, Z7, and Z23) variable-region EPIYA motifs for 6 h at 37°C before the cells were lysed and the samples were subjected to sodium dodecyl sulfate-polyacrylamide gel electrophoresis and Western blotting with antiphosphotyrosine monoclonal antibodies (lower panel). The blots were then stripped and reprobed with anti-CagA polyclonal antibodies (upper panel). (B) Densitometric analysis of the degree of CagA phosphorylation, expressed as a ratio of phospho-Tyr intensity to CagA protein intensity. The data shown are representative of those from three separate experiments.

    Journal: Journal of Clinical Microbiology

    Article Title: Tyrosine Phosphorylation of CagA from Chinese Helicobacter pylori Isolates in AGS Gastric Epithelial Cells

    doi: 10.1128/JCM.43.2.786-790.2005

    Figure Lengend Snippet: CagA with two variable-region TPMs is less phosphorylated than CagA with three variable-region TPMs. (A) AGS cells were cocultured with H. pylori strains possessing two (strain Z11) or three (strains Z4, Z7, and Z23) variable-region EPIYA motifs for 6 h at 37°C before the cells were lysed and the samples were subjected to sodium dodecyl sulfate-polyacrylamide gel electrophoresis and Western blotting with antiphosphotyrosine monoclonal antibodies (lower panel). The blots were then stripped and reprobed with anti-CagA polyclonal antibodies (upper panel). (B) Densitometric analysis of the degree of CagA phosphorylation, expressed as a ratio of phospho-Tyr intensity to CagA protein intensity. The data shown are representative of those from three separate experiments.

    Article Snippet: Samples were heated at 100°C for 5 min before analysis by sodium dodecyl sulfate-polyacrylamide gel electrophoresis and Western blotting with antiphosphotyrosine monoclonal antibodies (clone PY99; Santa Cruz Biotechnology, Santa Cruz, Calif.) or anti-CagA polyclonal antibodies (Austral Biologicals, San Ramon, Calif.).

    Techniques: Polyacrylamide Gel Electrophoresis, Western Blot

    Phosphorylation and expression of CagA protein following infection of gastric epithelial cells with representative H. pylori clinical isolates. CagA tyrosine phosphorylation (A) was evaluated by immunoblotting (IB) with anti-CagA rabbit polyclonal antibody

    Journal: Journal of Clinical Microbiology

    Article Title: CagA and VacA Polymorphisms Do Not Correlate with Severity of Histopathological Lesions in Helicobacter pylori-Infected Greek Children ▿-Infected Greek Children ▿ †

    doi: 10.1128/JCM.00159-09

    Figure Lengend Snippet: Phosphorylation and expression of CagA protein following infection of gastric epithelial cells with representative H. pylori clinical isolates. CagA tyrosine phosphorylation (A) was evaluated by immunoblotting (IB) with anti-CagA rabbit polyclonal antibody

    Article Snippet: CagA expression was detected by Western blot analysis using an anti-CagA primary polyclonal antibody (Austral Biologicals, San Ramon, CA) followed by a secondary horseradish peroxidase-conjugated goat anti-mouse immunoglobulin G polyclonal antibody (Jackson ImmunoResearch Europe, Ltd., Soham, Cambridgeshire, United Kingdom), detected by the ECL Plus chemiluminescence detection system (Amersham, GE Healthcare UK, Ltd., Buckinghamshire, United Kingdom).

    Techniques: Expressing, Infection

    Generation of phosphomimetic CagA mutants and their interaction with host signaling factors during H. pylori infection. ( A ) Site-directed Y > D mutagenesis of CagA EPIYA-A, EPIYA-B, and EPIYA-C to generate phosphomimetic mutants. Nontargeted tyrosines in adjacent EPIYA motifs were replaced by phenylalanines to avoid additional phosphorylation events per molecule. The resulting single, double, and triple mutants were named as indicated. ( B ) AGS cells were infected for 4 hours with CagA-expressing H. pylori strains as indicated. CagA phosphorylation was examined using α–PY-99 and α-CagA antibodies (arrows). All strains expressed similar amounts of CagA, but only H. pylori expressing WT CagA revealed a phosphorylation signal. The asterisk in the lower panel indicates antibody cross-reactivity with an unknown phosphorylated host cell protein. ( C – E ) AGS cells were infected for 4 hours with CagA-expressing H. pylori strains as indicated, and cell lysates were subjected to IP with α-CagA antibodies. Western blotting using α-Csk ( C ), α-PI3K ( D ), or α–SHP-2 ( E ) indicated that each of these factors formed a complex with the respective phosphomimetic CagA EPIYA Y > D mutant, but not with the nonphosphorylatable triple EPIYA-ABC Y > F mutant as control.

    Journal: The Journal of Clinical Investigation

    Article Title: c-Src and c-Abl kinases control hierarchic phosphorylation and function of the CagA effector protein in Western and East Asian Helicobacter pylori strains

    doi: 10.1172/JCI61143

    Figure Lengend Snippet: Generation of phosphomimetic CagA mutants and their interaction with host signaling factors during H. pylori infection. ( A ) Site-directed Y > D mutagenesis of CagA EPIYA-A, EPIYA-B, and EPIYA-C to generate phosphomimetic mutants. Nontargeted tyrosines in adjacent EPIYA motifs were replaced by phenylalanines to avoid additional phosphorylation events per molecule. The resulting single, double, and triple mutants were named as indicated. ( B ) AGS cells were infected for 4 hours with CagA-expressing H. pylori strains as indicated. CagA phosphorylation was examined using α–PY-99 and α-CagA antibodies (arrows). All strains expressed similar amounts of CagA, but only H. pylori expressing WT CagA revealed a phosphorylation signal. The asterisk in the lower panel indicates antibody cross-reactivity with an unknown phosphorylated host cell protein. ( C – E ) AGS cells were infected for 4 hours with CagA-expressing H. pylori strains as indicated, and cell lysates were subjected to IP with α-CagA antibodies. Western blotting using α-Csk ( C ), α-PI3K ( D ), or α–SHP-2 ( E ) indicated that each of these factors formed a complex with the respective phosphomimetic CagA EPIYA Y > D mutant, but not with the nonphosphorylatable triple EPIYA-ABC Y > F mutant as control.

    Article Snippet: Membranes were incubated with mouse monoclonal α-phosphotyrosine antibody PY-99 (Santa Cruz) or monoclonal anti-CagA antibody (Austral Biologicals).

    Techniques: Infection, Mutagenesis, Expressing, Western Blot

    Dual infection of H. pylori strains expressing different single phosphorylatable or phosphomimetic EPIYA motifs induces AGS cell elongation. AGS cells were infected for 4 hours with CagA EPIYA Y > F ( A ) or EPIYA Y > D ( B ) mutant strains as indicated. The available single phosphorylatable or phosphomimetic EPIYA motifs for each double infection are indicated. The number of elongated cells in each experiment was quantitated in triplicate in 10 different 0.25-mm 2 fields, and CagA phosphorylation was examined using α–PY-99 and α-CagA antibodies (arrows). ( C ) AGS infection for 4 hours with the indicated CagA-expressing strains in the presence or absence of c-Src inhibitor PP2 (10 μM) or c-Abl inhibitor SKI-DV2-43 (1 μM) revealed significant changes in AGS cell elongation as quantitated in triplicate in 10 different 0.25-mm 2 fields. * P ≤ 0.01; ** P ≤ 0.001.

    Journal: The Journal of Clinical Investigation

    Article Title: c-Src and c-Abl kinases control hierarchic phosphorylation and function of the CagA effector protein in Western and East Asian Helicobacter pylori strains

    doi: 10.1172/JCI61143

    Figure Lengend Snippet: Dual infection of H. pylori strains expressing different single phosphorylatable or phosphomimetic EPIYA motifs induces AGS cell elongation. AGS cells were infected for 4 hours with CagA EPIYA Y > F ( A ) or EPIYA Y > D ( B ) mutant strains as indicated. The available single phosphorylatable or phosphomimetic EPIYA motifs for each double infection are indicated. The number of elongated cells in each experiment was quantitated in triplicate in 10 different 0.25-mm 2 fields, and CagA phosphorylation was examined using α–PY-99 and α-CagA antibodies (arrows). ( C ) AGS infection for 4 hours with the indicated CagA-expressing strains in the presence or absence of c-Src inhibitor PP2 (10 μM) or c-Abl inhibitor SKI-DV2-43 (1 μM) revealed significant changes in AGS cell elongation as quantitated in triplicate in 10 different 0.25-mm 2 fields. * P ≤ 0.01; ** P ≤ 0.001.

    Article Snippet: Membranes were incubated with mouse monoclonal α-phosphotyrosine antibody PY-99 (Santa Cruz) or monoclonal anti-CagA antibody (Austral Biologicals).

    Techniques: Infection, Expressing, Mutagenesis

    In vitro phosphorylation of CagA mutants by c-Abl or c-Src kinases. ( A ) Site-directed Y > F mutagenesis of CagA EPIYA-A, EPIYA-B, and EPIYA-C in strain 26695. Single, double, and triple mutants were named as indicated. ( B and C ) Lysates of H. pylori expressing the mutated CagA EPIYA motifs were subjected to in vitro phosphorylation assays using recombinant c-Src kinase ( B ) or c-Abl kinase ( C ). Immunoblotting using α–PY-99 and α-CagA antibodies (arrows) indicated that both c-Src and c-Abl phosphorylated CagA in a different fashion. ( D ) Schematic diagram of the data, showing that c-Src only phosphorylated Y-972 in EPIYA-C, while Abl can phosphorylate Y-899, Y-918, and Y-972 in EPIYA-A, EPIYA-B, and EPIYA-C, respectively.

    Journal: The Journal of Clinical Investigation

    Article Title: c-Src and c-Abl kinases control hierarchic phosphorylation and function of the CagA effector protein in Western and East Asian Helicobacter pylori strains

    doi: 10.1172/JCI61143

    Figure Lengend Snippet: In vitro phosphorylation of CagA mutants by c-Abl or c-Src kinases. ( A ) Site-directed Y > F mutagenesis of CagA EPIYA-A, EPIYA-B, and EPIYA-C in strain 26695. Single, double, and triple mutants were named as indicated. ( B and C ) Lysates of H. pylori expressing the mutated CagA EPIYA motifs were subjected to in vitro phosphorylation assays using recombinant c-Src kinase ( B ) or c-Abl kinase ( C ). Immunoblotting using α–PY-99 and α-CagA antibodies (arrows) indicated that both c-Src and c-Abl phosphorylated CagA in a different fashion. ( D ) Schematic diagram of the data, showing that c-Src only phosphorylated Y-972 in EPIYA-C, while Abl can phosphorylate Y-899, Y-918, and Y-972 in EPIYA-A, EPIYA-B, and EPIYA-C, respectively.

    Article Snippet: Membranes were incubated with mouse monoclonal α-phosphotyrosine antibody PY-99 (Santa Cruz) or monoclonal anti-CagA antibody (Austral Biologicals).

    Techniques: In Vitro, Mutagenesis, Expressing, Recombinant

    Role of EPIYA motifs in CagA phosphorylation and AGS cell elongation during H. pylori infection. ( A ) AGS cells were infected for 4 hours with CagA-expressing H. pylori strains as indicated. Phosphorylation of CagA was examined using α–PY-99 and α-CagA antibodies (arrows). ( B ) The number of elongated cells in each experiment was quantitated in triplicate in 10 different 0.25-mm 2 fields. ( C ) Phase-contrast micrographs of AGS cells infected with the different strains as indicated. ** P ≤ 0.001.

    Journal: The Journal of Clinical Investigation

    Article Title: c-Src and c-Abl kinases control hierarchic phosphorylation and function of the CagA effector protein in Western and East Asian Helicobacter pylori strains

    doi: 10.1172/JCI61143

    Figure Lengend Snippet: Role of EPIYA motifs in CagA phosphorylation and AGS cell elongation during H. pylori infection. ( A ) AGS cells were infected for 4 hours with CagA-expressing H. pylori strains as indicated. Phosphorylation of CagA was examined using α–PY-99 and α-CagA antibodies (arrows). ( B ) The number of elongated cells in each experiment was quantitated in triplicate in 10 different 0.25-mm 2 fields. ( C ) Phase-contrast micrographs of AGS cells infected with the different strains as indicated. ** P ≤ 0.001.

    Article Snippet: Membranes were incubated with mouse monoclonal α-phosphotyrosine antibody PY-99 (Santa Cruz) or monoclonal anti-CagA antibody (Austral Biologicals).

    Techniques: Infection, Expressing

    Analysis of CagA PY protein species during infection with H. pylori by 1-DE and 2-DE. ( A ). ( B ) AGS cells were infected for the indicated times with strain 26695. The resulting protein lysates were separated by 1-DE, and phosphorylation of injected CagA was examined using α–PY-99 and α-CagA antibodies (arrows). ( C ) Separation of CagA protein species from B by 2-DE. Depending on the time of infection, full-length CagA PY appeared as 1 spot (spot 1, red arrows, pI = 7.0) or 2 spots (spots 1 and 2; spot 2, green arrows, pI = 6.5) as indicated. The α-CagA antibody probe revealed a third spot (spot 3, blue arrows, pI = 7.5). Overlay of both exposures yielded 2 or 3 spots as shown. Strain TN2-GF4 exhibited the same pattern as 26695 (bottom). ( D ) Inhibition of Src with PP2 (10 μM) or Abl with SKI-DV2-43 (1 μM) revealed significant changes in spot intensity depending on the time of infection.

    Journal: The Journal of Clinical Investigation

    Article Title: c-Src and c-Abl kinases control hierarchic phosphorylation and function of the CagA effector protein in Western and East Asian Helicobacter pylori strains

    doi: 10.1172/JCI61143

    Figure Lengend Snippet: Analysis of CagA PY protein species during infection with H. pylori by 1-DE and 2-DE. ( A ). ( B ) AGS cells were infected for the indicated times with strain 26695. The resulting protein lysates were separated by 1-DE, and phosphorylation of injected CagA was examined using α–PY-99 and α-CagA antibodies (arrows). ( C ) Separation of CagA protein species from B by 2-DE. Depending on the time of infection, full-length CagA PY appeared as 1 spot (spot 1, red arrows, pI = 7.0) or 2 spots (spots 1 and 2; spot 2, green arrows, pI = 6.5) as indicated. The α-CagA antibody probe revealed a third spot (spot 3, blue arrows, pI = 7.5). Overlay of both exposures yielded 2 or 3 spots as shown. Strain TN2-GF4 exhibited the same pattern as 26695 (bottom). ( D ) Inhibition of Src with PP2 (10 μM) or Abl with SKI-DV2-43 (1 μM) revealed significant changes in spot intensity depending on the time of infection.

    Article Snippet: Membranes were incubated with mouse monoclonal α-phosphotyrosine antibody PY-99 (Santa Cruz) or monoclonal anti-CagA antibody (Austral Biologicals).

    Techniques: Infection, Injection, Inhibition

    Sequence comparison of the three TPM sites in CagA proteins from different clinical H. pylori strains and specific detection of phosphorylated EPIYT-motif during co-culture. Panel A : CagA proteins of H. pylori vary in their carboxy-terminal TPM sites. These EPIYA-repeats serve as tyrosine phosphorylation sites of CagA and can be targeted by c-Abl and c-Src kinases. Three EPIYA- or EPIYT-segments at position A, B and C are shaded with yellow. One striking feature of B-TPM is the presence of a threonine residue in the +1 position (shaded with blue) relative to the phosphorylated tyrosine residue, which is highly conserved in most but not all H. pylori strains and may affect the capabilities of binding the p85 subunit of PI3-kinase, as discussed in the text. The CagA protein sequences were obtained from databases and sequence alignment was done using the ClustalW2 program ( http://www.ebi.ac.uk/Tools/msa/clustalw2/ ). Panel B : To investigate whether the EPIYT-motif can be phosphorylated during co-culture, AGS cells were co-incubated with the indicated CagA-expressing H. pylori strains for 6 h. Phosphorylation of CagA was examined using the phospho-specific α-pCagA-EPIYT-918 antibody. Loading of equal amounts of protein in each sample was confirmed by probing with monoclonal α-CagA and α-GAPDH antibodies.

    Journal: PLoS Pathogens

    Article Title: A Specific A/T Polymorphism in Western Tyrosine Phosphorylation B-Motifs Regulates Helicobacter pylori CagA Epithelial Cell Interactions

    doi: 10.1371/journal.ppat.1004621

    Figure Lengend Snippet: Sequence comparison of the three TPM sites in CagA proteins from different clinical H. pylori strains and specific detection of phosphorylated EPIYT-motif during co-culture. Panel A : CagA proteins of H. pylori vary in their carboxy-terminal TPM sites. These EPIYA-repeats serve as tyrosine phosphorylation sites of CagA and can be targeted by c-Abl and c-Src kinases. Three EPIYA- or EPIYT-segments at position A, B and C are shaded with yellow. One striking feature of B-TPM is the presence of a threonine residue in the +1 position (shaded with blue) relative to the phosphorylated tyrosine residue, which is highly conserved in most but not all H. pylori strains and may affect the capabilities of binding the p85 subunit of PI3-kinase, as discussed in the text. The CagA protein sequences were obtained from databases and sequence alignment was done using the ClustalW2 program ( http://www.ebi.ac.uk/Tools/msa/clustalw2/ ). Panel B : To investigate whether the EPIYT-motif can be phosphorylated during co-culture, AGS cells were co-incubated with the indicated CagA-expressing H. pylori strains for 6 h. Phosphorylation of CagA was examined using the phospho-specific α-pCagA-EPIYT-918 antibody. Loading of equal amounts of protein in each sample was confirmed by probing with monoclonal α-CagA and α-GAPDH antibodies.

    Article Snippet: Two micrograms of the monoclonal α-CagA antibody (Austral Biologicals, San Ramon CA) or polyclonal antibody against the p85 subunit of PI3-kinase (Santa Cruz Biotechnology, Dallas TX) were added to the supernatant and incubated overnight at 4°C on a shaker.

    Techniques: Sequencing, Co-Culture Assay, Binding Assay, Incubation, Expressing

    The EPIYT site at B-TPM of CagA is phosphorylated and necessary for interaction with PI3-kinase. Panel A : Site-directed mutagenesis of CagA TPM-motifs A, B and C was performed to generate the indicated phospho-resistant variants. Tyrosine residues in adjacent TPM-motifs were replaced by phenylalanines. The resulting single and double mutants are indicated and complemented into the H. pylori Δ cagA mutant. Panel B : AGS cells were co-cultured with the various CagA-expressing H. pylori strains for 6 h as indicated. Cell extracts were harvested and subjected to immunoprecipitation (IP) using α-CagA antibodies. CagA phosphorylation in the IPs was examined using α-pY-99 and α-CagA antibodies (arrows). All strains expressed similar amounts of CagA, and H. pylori expressing CagA wild-type (wt), EPIYT-AC Y > F , and EPIYT-B Y > F all showed phosphorylation signal. Western blotting using α-PI3-kinase antibody revealed that only CagA wt and EPIYT-AC Y > F can bind to PI3-kinase, but not the EPIYT-B Y > F mutant, suggesting that EPIYT-B is phosphorylated and necessary for the interaction.

    Journal: PLoS Pathogens

    Article Title: A Specific A/T Polymorphism in Western Tyrosine Phosphorylation B-Motifs Regulates Helicobacter pylori CagA Epithelial Cell Interactions

    doi: 10.1371/journal.ppat.1004621

    Figure Lengend Snippet: The EPIYT site at B-TPM of CagA is phosphorylated and necessary for interaction with PI3-kinase. Panel A : Site-directed mutagenesis of CagA TPM-motifs A, B and C was performed to generate the indicated phospho-resistant variants. Tyrosine residues in adjacent TPM-motifs were replaced by phenylalanines. The resulting single and double mutants are indicated and complemented into the H. pylori Δ cagA mutant. Panel B : AGS cells were co-cultured with the various CagA-expressing H. pylori strains for 6 h as indicated. Cell extracts were harvested and subjected to immunoprecipitation (IP) using α-CagA antibodies. CagA phosphorylation in the IPs was examined using α-pY-99 and α-CagA antibodies (arrows). All strains expressed similar amounts of CagA, and H. pylori expressing CagA wild-type (wt), EPIYT-AC Y > F , and EPIYT-B Y > F all showed phosphorylation signal. Western blotting using α-PI3-kinase antibody revealed that only CagA wt and EPIYT-AC Y > F can bind to PI3-kinase, but not the EPIYT-B Y > F mutant, suggesting that EPIYT-B is phosphorylated and necessary for the interaction.

    Article Snippet: Two micrograms of the monoclonal α-CagA antibody (Austral Biologicals, San Ramon CA) or polyclonal antibody against the p85 subunit of PI3-kinase (Santa Cruz Biotechnology, Dallas TX) were added to the supernatant and incubated overnight at 4°C on a shaker.

    Techniques: Mutagenesis, Cell Culture, Expressing, Immunoprecipitation, Western Blot

    PI3-kinase can interact with B-TPM of CagA during co-culture. Panel A : Site-directed mutagenesis of CagA TPM-motifs A, B and C was performed to generate the indicated phospho-resistant variants. Tyrosine residues in adjacent TPM-motifs were replaced by phenylalanines. The resulting single and triple mutants were named as indicated and complemented into the H. pylori Δ cagA mutant. Panel B : AGS cells were co-cultured with the various CagA-expressing H. pylori strains for 6 h as indicated. Cell extracts were harvested and subjected to reverse immunoprecipitation (IP) using α-PI3-kinase antibodies. All samples contained similar amounts of PI3-kinase in the input control. CagA presence and phosphorylation at the EPIYT-site in the IPs was examined using phospho-specific α-pCagA-EPIYT-918 and α-CagA antibodies (arrows). Only the lane with H. pylori expressing CagA wt revealed a signal for CagA and phosphorylation at EPIYT-918 in the IP, indicating that phosphorylated EPIYT-B is necessary for the interaction with PI3-kinase. Panel C : After 24 h co-culture of AGS cells with the isogenic H. pylori strains containing the engineered CagA molecules, whole cell lysates were subjected to immunoprecipitation with an anti-CagA antibody. The anti-CagA immunoprecipitates (IP) were separated on SDS-PAGE, followed by western blot with anti-PI3-kinase (p85), which indicated that the engineered CagA B-EPIYA and CagA B-EPIYT molecules have different affinity to the PI3-kinase protein in AGS cells.

    Journal: PLoS Pathogens

    Article Title: A Specific A/T Polymorphism in Western Tyrosine Phosphorylation B-Motifs Regulates Helicobacter pylori CagA Epithelial Cell Interactions

    doi: 10.1371/journal.ppat.1004621

    Figure Lengend Snippet: PI3-kinase can interact with B-TPM of CagA during co-culture. Panel A : Site-directed mutagenesis of CagA TPM-motifs A, B and C was performed to generate the indicated phospho-resistant variants. Tyrosine residues in adjacent TPM-motifs were replaced by phenylalanines. The resulting single and triple mutants were named as indicated and complemented into the H. pylori Δ cagA mutant. Panel B : AGS cells were co-cultured with the various CagA-expressing H. pylori strains for 6 h as indicated. Cell extracts were harvested and subjected to reverse immunoprecipitation (IP) using α-PI3-kinase antibodies. All samples contained similar amounts of PI3-kinase in the input control. CagA presence and phosphorylation at the EPIYT-site in the IPs was examined using phospho-specific α-pCagA-EPIYT-918 and α-CagA antibodies (arrows). Only the lane with H. pylori expressing CagA wt revealed a signal for CagA and phosphorylation at EPIYT-918 in the IP, indicating that phosphorylated EPIYT-B is necessary for the interaction with PI3-kinase. Panel C : After 24 h co-culture of AGS cells with the isogenic H. pylori strains containing the engineered CagA molecules, whole cell lysates were subjected to immunoprecipitation with an anti-CagA antibody. The anti-CagA immunoprecipitates (IP) were separated on SDS-PAGE, followed by western blot with anti-PI3-kinase (p85), which indicated that the engineered CagA B-EPIYA and CagA B-EPIYT molecules have different affinity to the PI3-kinase protein in AGS cells.

    Article Snippet: Two micrograms of the monoclonal α-CagA antibody (Austral Biologicals, San Ramon CA) or polyclonal antibody against the p85 subunit of PI3-kinase (Santa Cruz Biotechnology, Dallas TX) were added to the supernatant and incubated overnight at 4°C on a shaker.

    Techniques: Co-Culture Assay, Mutagenesis, Cell Culture, Expressing, Immunoprecipitation, SDS Page, Western Blot

    Sequence comparison of the three TPM sites in CagA proteins from different clinical H. pylori strains and specific detection of phosphorylated EPIYT-motif during co-culture. Panel A : CagA proteins of H. pylori vary in their carboxy-terminal TPM sites. These EPIYA-repeats serve as tyrosine phosphorylation sites of CagA and can be targeted by c-Abl and c-Src kinases. Three EPIYA- or EPIYT-segments at position A, B and C are shaded with yellow. One striking feature of B-TPM is the presence of a threonine residue in the +1 position (shaded with blue) relative to the phosphorylated tyrosine residue, which is highly conserved in most but not all H. pylori strains and may affect the capabilities of binding the p85 subunit of PI3-kinase, as discussed in the text. The CagA protein sequences were obtained from databases and sequence alignment was done using the ClustalW2 program ( http://www.ebi.ac.uk/Tools/msa/clustalw2/ ). Panel B : To investigate whether the EPIYT-motif can be phosphorylated during co-culture, AGS cells were co-incubated with the indicated CagA-expressing H. pylori strains for 6 h. Phosphorylation of CagA was examined using the phospho-specific α-pCagA-EPIYT-918 antibody. Loading of equal amounts of protein in each sample was confirmed by probing with monoclonal α-CagA and α-GAPDH antibodies.

    Journal: PLoS Pathogens

    Article Title: A Specific A/T Polymorphism in Western Tyrosine Phosphorylation B-Motifs Regulates Helicobacter pylori CagA Epithelial Cell Interactions

    doi: 10.1371/journal.ppat.1004621

    Figure Lengend Snippet: Sequence comparison of the three TPM sites in CagA proteins from different clinical H. pylori strains and specific detection of phosphorylated EPIYT-motif during co-culture. Panel A : CagA proteins of H. pylori vary in their carboxy-terminal TPM sites. These EPIYA-repeats serve as tyrosine phosphorylation sites of CagA and can be targeted by c-Abl and c-Src kinases. Three EPIYA- or EPIYT-segments at position A, B and C are shaded with yellow. One striking feature of B-TPM is the presence of a threonine residue in the +1 position (shaded with blue) relative to the phosphorylated tyrosine residue, which is highly conserved in most but not all H. pylori strains and may affect the capabilities of binding the p85 subunit of PI3-kinase, as discussed in the text. The CagA protein sequences were obtained from databases and sequence alignment was done using the ClustalW2 program ( http://www.ebi.ac.uk/Tools/msa/clustalw2/ ). Panel B : To investigate whether the EPIYT-motif can be phosphorylated during co-culture, AGS cells were co-incubated with the indicated CagA-expressing H. pylori strains for 6 h. Phosphorylation of CagA was examined using the phospho-specific α-pCagA-EPIYT-918 antibody. Loading of equal amounts of protein in each sample was confirmed by probing with monoclonal α-CagA and α-GAPDH antibodies.

    Article Snippet: Rabbit polyclonal and mouse monoclonal α-CagA antibodies were from Austral Biologicals, or from Emd Millipore Corporation (Billerica MA).

    Techniques: Sequencing, Co-Culture Assay, Binding Assay, Incubation, Expressing

    The EPIYT site at B-TPM of CagA is phosphorylated and necessary for interaction with PI3-kinase. Panel A : Site-directed mutagenesis of CagA TPM-motifs A, B and C was performed to generate the indicated phospho-resistant variants. Tyrosine residues in adjacent TPM-motifs were replaced by phenylalanines. The resulting single and double mutants are indicated and complemented into the H. pylori Δ cagA mutant. Panel B : AGS cells were co-cultured with the various CagA-expressing H. pylori strains for 6 h as indicated. Cell extracts were harvested and subjected to immunoprecipitation (IP) using α-CagA antibodies. CagA phosphorylation in the IPs was examined using α-pY-99 and α-CagA antibodies (arrows). All strains expressed similar amounts of CagA, and H. pylori expressing CagA wild-type (wt), EPIYT-AC Y > F , and EPIYT-B Y > F all showed phosphorylation signal. Western blotting using α-PI3-kinase antibody revealed that only CagA wt and EPIYT-AC Y > F can bind to PI3-kinase, but not the EPIYT-B Y > F mutant, suggesting that EPIYT-B is phosphorylated and necessary for the interaction.

    Journal: PLoS Pathogens

    Article Title: A Specific A/T Polymorphism in Western Tyrosine Phosphorylation B-Motifs Regulates Helicobacter pylori CagA Epithelial Cell Interactions

    doi: 10.1371/journal.ppat.1004621

    Figure Lengend Snippet: The EPIYT site at B-TPM of CagA is phosphorylated and necessary for interaction with PI3-kinase. Panel A : Site-directed mutagenesis of CagA TPM-motifs A, B and C was performed to generate the indicated phospho-resistant variants. Tyrosine residues in adjacent TPM-motifs were replaced by phenylalanines. The resulting single and double mutants are indicated and complemented into the H. pylori Δ cagA mutant. Panel B : AGS cells were co-cultured with the various CagA-expressing H. pylori strains for 6 h as indicated. Cell extracts were harvested and subjected to immunoprecipitation (IP) using α-CagA antibodies. CagA phosphorylation in the IPs was examined using α-pY-99 and α-CagA antibodies (arrows). All strains expressed similar amounts of CagA, and H. pylori expressing CagA wild-type (wt), EPIYT-AC Y > F , and EPIYT-B Y > F all showed phosphorylation signal. Western blotting using α-PI3-kinase antibody revealed that only CagA wt and EPIYT-AC Y > F can bind to PI3-kinase, but not the EPIYT-B Y > F mutant, suggesting that EPIYT-B is phosphorylated and necessary for the interaction.

    Article Snippet: Rabbit polyclonal and mouse monoclonal α-CagA antibodies were from Austral Biologicals, or from Emd Millipore Corporation (Billerica MA).

    Techniques: Mutagenesis, Cell Culture, Expressing, Immunoprecipitation, Western Blot

    PI3-kinase can interact with B-TPM of CagA during co-culture. Panel A : Site-directed mutagenesis of CagA TPM-motifs A, B and C was performed to generate the indicated phospho-resistant variants. Tyrosine residues in adjacent TPM-motifs were replaced by phenylalanines. The resulting single and triple mutants were named as indicated and complemented into the H. pylori Δ cagA mutant. Panel B : AGS cells were co-cultured with the various CagA-expressing H. pylori strains for 6 h as indicated. Cell extracts were harvested and subjected to reverse immunoprecipitation (IP) using α-PI3-kinase antibodies. All samples contained similar amounts of PI3-kinase in the input control. CagA presence and phosphorylation at the EPIYT-site in the IPs was examined using phospho-specific α-pCagA-EPIYT-918 and α-CagA antibodies (arrows). Only the lane with H. pylori expressing CagA wt revealed a signal for CagA and phosphorylation at EPIYT-918 in the IP, indicating that phosphorylated EPIYT-B is necessary for the interaction with PI3-kinase. Panel C : After 24 h co-culture of AGS cells with the isogenic H. pylori strains containing the engineered CagA molecules, whole cell lysates were subjected to immunoprecipitation with an anti-CagA antibody. The anti-CagA immunoprecipitates (IP) were separated on SDS-PAGE, followed by western blot with anti-PI3-kinase (p85), which indicated that the engineered CagA B-EPIYA and CagA B-EPIYT molecules have different affinity to the PI3-kinase protein in AGS cells.

    Journal: PLoS Pathogens

    Article Title: A Specific A/T Polymorphism in Western Tyrosine Phosphorylation B-Motifs Regulates Helicobacter pylori CagA Epithelial Cell Interactions

    doi: 10.1371/journal.ppat.1004621

    Figure Lengend Snippet: PI3-kinase can interact with B-TPM of CagA during co-culture. Panel A : Site-directed mutagenesis of CagA TPM-motifs A, B and C was performed to generate the indicated phospho-resistant variants. Tyrosine residues in adjacent TPM-motifs were replaced by phenylalanines. The resulting single and triple mutants were named as indicated and complemented into the H. pylori Δ cagA mutant. Panel B : AGS cells were co-cultured with the various CagA-expressing H. pylori strains for 6 h as indicated. Cell extracts were harvested and subjected to reverse immunoprecipitation (IP) using α-PI3-kinase antibodies. All samples contained similar amounts of PI3-kinase in the input control. CagA presence and phosphorylation at the EPIYT-site in the IPs was examined using phospho-specific α-pCagA-EPIYT-918 and α-CagA antibodies (arrows). Only the lane with H. pylori expressing CagA wt revealed a signal for CagA and phosphorylation at EPIYT-918 in the IP, indicating that phosphorylated EPIYT-B is necessary for the interaction with PI3-kinase. Panel C : After 24 h co-culture of AGS cells with the isogenic H. pylori strains containing the engineered CagA molecules, whole cell lysates were subjected to immunoprecipitation with an anti-CagA antibody. The anti-CagA immunoprecipitates (IP) were separated on SDS-PAGE, followed by western blot with anti-PI3-kinase (p85), which indicated that the engineered CagA B-EPIYA and CagA B-EPIYT molecules have different affinity to the PI3-kinase protein in AGS cells.

    Article Snippet: Rabbit polyclonal and mouse monoclonal α-CagA antibodies were from Austral Biologicals, or from Emd Millipore Corporation (Billerica MA).

    Techniques: Co-Culture Assay, Mutagenesis, Cell Culture, Expressing, Immunoprecipitation, SDS Page, Western Blot