anti-caga antibodies Search Results


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  • 89
    Santa Cruz Biotechnology anti caga antibody
    Variation in the <t>CagA</t> amino acid sequence of East Asian-type and Western-type CagA. The target <t>α-EAS</t> sequences “AINRKIDRINKIASAGKG” in EPIYA segment D, based on the sequences of Japanese strains, are shown in frame. A sequence analysis revealed that the typical target sequences in Bhutanese strains was “(T/K)IN(G/R)KID(Q/R)(L/I)N(R/K)(T/I)ASA(G/N)KG,” where (X/Y) means X and Y as the two major amino acids. The sequences of the EPIYA-D segments in Bhutanese strains were highly variable compared to strains deposited in GenBank. In contrast, the East Asian-type and Western-type sequences of EPIYA-C segments in Bhutanese strains were largely identical, similar to the typical sequences of EPIYA-C segments deposited in GenBank. Reference strains used in Fig. 1A (strain name [accession number]) were 103a (AB110966.1), 105a (AB110967.1), 106a (AB110968.1), 108a (AB110969.1), 113b (AB110970.1), 120a (AB110971.1), 122b (AB110972.1), 125b (AB110973.1), 128a (AB110974.1), FJT77 (KF028580.1), 04-518 (AB267252.1), 03-166 (AB267253.1), 04-264 (AB267254.1), THP1477 (AB116744.1), 04-334 (AB267249.1), 03-292 (AB267250.1), 04-366 (AB267251.1), THP1260 (AB116742.1), M3 (AB116740.1), THP463 (AB116735.1), Korea23 (AB057044.1), Korea 12 (AB057043.1), K69 (FJ458129.1), Korea2-3 (AB057040.1), k266 (FJ458163.1), K265 (FJ458162.1), K264 (FJ458161.1), K261 (FJ458158.1), K260 (FJ458157.1), K259 (FJ458156.1). Reference strains used in Fig. 1C (strain name [accession number]) were India41 (AF222807.1), India99 (AF222809.1), OSC40A (EU089774.1), OSC42B (EU089775.1), PCR-156i (EU368669.1), PCR218vi (EU089766.1). RIGLD-OC149 (JX428784.1), SAN53 (EU089771.1), PD682 (EF450167.1), PD636 (EF450165.1), PD308 (EF450162.1), PD488 (EF450161.1), PD537 (EF450160.1), PD501 (EF450159.1), PD351 (EF450158.1), PD348 (EF450157.1), PD6481K (EF450153.1), 216G (GQ899171.1), 1407 (GU143415.1), HPI-14 (FJ849792.1), HPI-13 (FJ849791.1), HPI-11 (FJ849789.1), USA2791 (AB057099.1), Kazak3 (AB057098.1), USA35 (AB057095.1), Italy329 (AB057094.1), Arizona2 (AB057075.1), Arizona1 (AB057074.1).
    Anti Caga Antibody, supplied by Santa Cruz Biotechnology, used in various techniques. Bioz Stars score: 89/100, based on 70 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Austral Biologicals rabbit polyclonal α caga antibody
    <t>CagA</t> injection by H. pylori cannot be achieved in infected HEK293 cell lines but in AGS gastric epithelial cells. Western blot analysis of (A) AGS, (B) HEK293-TLR2, (C) HEK293 and (D) HEK293-TLR5 cells infected with H. pylori wild-type strains P1, P12, P310 or 26695 for 6 hours. Phosphorylation of injected CagA was monitored using phosphotyrosine α-PY-99 and <t>α-CagA</t> antibodies. Red arrows indicate the position of phosphorylated CagA on the blot. Western blots for the house keeping gene GAPDH served as loading control.
    Rabbit Polyclonal α Caga Antibody, supplied by Austral Biologicals, used in various techniques. Bioz Stars score: 88/100, based on 37 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Austral Biologicals anti caga antibodies
    PC activation induced by <t>VacA</t> and <t>CagA</t> on THP-1 cells. Both VacA and CagA significantly ( P
    Anti Caga Antibodies, supplied by Austral Biologicals, used in various techniques. Bioz Stars score: 90/100, based on 5 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    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 6 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.
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    Santa Cruz Biotechnology mouse anti caga antibody
    Ubiquitination of total proteins and proteasome activity in AGS cells co-cultured with H. pylori . (A) Representative immunoblot of total ubiquitinated protein from AGS cells infected with H. pylori (MOI = 300) for 8 h, in presence or absence of 2.5 μM Lactacystin. <t>Tubulin</t> is shown as loading control and <t>CagA</t> as infection control. (B) Quantification of proteasome activity in AGS cells infected with H. pylori wild type and its isogenic mutants cagA, vacA and, in presence of 2.5 μM Lactacystin for 8 h. Asterisk represents significant difference ( p
    Mouse Anti Caga Antibody, supplied by Santa Cruz Biotechnology, used in various techniques. Bioz Stars score: 93/100, based on 16 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Santa Cruz Biotechnology polyclonal anti caga antibody
    Analysis of the <t>CagA</t> protein patterns by immunoblot analysis. (A) Lysates of H . pylori isolated from the tissue specimens of various gastric patients were prepared as described in the Materials and Methods. Aliquots of protein (20 μg) were separated by SDS-gel electrophoresis and CagA was detected by immunoblotting using <t>polyclonal</t> anti-CagA antibody b-300 (Santa Cruz). The H . pylori 60190 and 26695 cagA knockout mutant (26695ΔA) were included as a positive and negative control, respectively, for CagA expression. The numbers at the top identify the various H . pylori colonies, while those on the left indicate the molecular weight markers in kDa. It can be seen that there are four types of CagA fragmentation patterns. (B) The percentages of ABD subtypes based on the CagA protein fragmentation patterns across the 96 H . pylori cagA ABD colonies that were analyzed and clearly ABD-2 is the predominant type.
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    Abcam anti caga antibody
    Analysis of the <t>CagA</t> protein patterns by immunoblot analysis. (A) Lysates of H . pylori isolated from the tissue specimens of various gastric patients were prepared as described in the Materials and Methods. Aliquots of protein (20 μg) were separated by SDS-gel electrophoresis and CagA was detected by immunoblotting using <t>polyclonal</t> anti-CagA antibody b-300 (Santa Cruz). The H . pylori 60190 and 26695 cagA knockout mutant (26695ΔA) were included as a positive and negative control, respectively, for CagA expression. The numbers at the top identify the various H . pylori colonies, while those on the left indicate the molecular weight markers in kDa. It can be seen that there are four types of CagA fragmentation patterns. (B) The percentages of ABD subtypes based on the CagA protein fragmentation patterns across the 96 H . pylori cagA ABD colonies that were analyzed and clearly ABD-2 is the predominant type.
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    Santa Cruz Biotechnology rabbit polyclonal anti caga antibody b 300
    Analysis of the <t>CagA</t> protein patterns by immunoblot analysis. (A) Lysates of H . pylori isolated from the tissue specimens of various gastric patients were prepared as described in the Materials and Methods. Aliquots of protein (20 μg) were separated by SDS-gel electrophoresis and CagA was detected by immunoblotting using <t>polyclonal</t> anti-CagA antibody b-300 (Santa Cruz). The H . pylori 60190 and 26695 cagA knockout mutant (26695ΔA) were included as a positive and negative control, respectively, for CagA expression. The numbers at the top identify the various H . pylori colonies, while those on the left indicate the molecular weight markers in kDa. It can be seen that there are four types of CagA fragmentation patterns. (B) The percentages of ABD subtypes based on the CagA protein fragmentation patterns across the 96 H . pylori cagA ABD colonies that were analyzed and clearly ABD-2 is the predominant type.
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    Santa Cruz Biotechnology anti caga monoclonal antibody
    Analysis of the <t>CagA</t> protein patterns by immunoblot analysis. (A) Lysates of H . pylori isolated from the tissue specimens of various gastric patients were prepared as described in the Materials and Methods. Aliquots of protein (20 μg) were separated by SDS-gel electrophoresis and CagA was detected by immunoblotting using <t>polyclonal</t> anti-CagA antibody b-300 (Santa Cruz). The H . pylori 60190 and 26695 cagA knockout mutant (26695ΔA) were included as a positive and negative control, respectively, for CagA expression. The numbers at the top identify the various H . pylori colonies, while those on the left indicate the molecular weight markers in kDa. It can be seen that there are four types of CagA fragmentation patterns. (B) The percentages of ABD subtypes based on the CagA protein fragmentation patterns across the 96 H . pylori cagA ABD colonies that were analyzed and clearly ABD-2 is the predominant type.
    Anti Caga Monoclonal Antibody, supplied by Santa Cruz Biotechnology, used in various techniques. Bioz Stars score: 85/100, based on 12 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    92
    Santa Cruz Biotechnology anti caga rabbit polyclonal antibody
    Analysis of the <t>CagA</t> protein patterns by immunoblot analysis. (A) Lysates of H . pylori isolated from the tissue specimens of various gastric patients were prepared as described in the Materials and Methods. Aliquots of protein (20 μg) were separated by SDS-gel electrophoresis and CagA was detected by immunoblotting using <t>polyclonal</t> anti-CagA antibody b-300 (Santa Cruz). The H . pylori 60190 and 26695 cagA knockout mutant (26695ΔA) were included as a positive and negative control, respectively, for CagA expression. The numbers at the top identify the various H . pylori colonies, while those on the left indicate the molecular weight markers in kDa. It can be seen that there are four types of CagA fragmentation patterns. (B) The percentages of ABD subtypes based on the CagA protein fragmentation patterns across the 96 H . pylori cagA ABD colonies that were analyzed and clearly ABD-2 is the predominant type.
    Anti Caga Rabbit Polyclonal Antibody, supplied by Santa Cruz Biotechnology, 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
    Analysis of the <t>CagA</t> protein patterns by immunoblot analysis. (A) Lysates of H . pylori isolated from the tissue specimens of various gastric patients were prepared as described in the Materials and Methods. Aliquots of protein (20 μg) were separated by SDS-gel electrophoresis and CagA was detected by immunoblotting using <t>polyclonal</t> anti-CagA antibody b-300 (Santa Cruz). The H . pylori 60190 and 26695 cagA knockout mutant (26695ΔA) were included as a positive and negative control, respectively, for CagA expression. The numbers at the top identify the various H . pylori colonies, while those on the left indicate the molecular weight markers in kDa. It can be seen that there are four types of CagA fragmentation patterns. (B) The percentages of ABD subtypes based on the CagA protein fragmentation patterns across the 96 H . pylori cagA ABD colonies that were analyzed and clearly ABD-2 is the predominant type.
    Monoclonal Anti Caga Antibody, supplied by Abcam, used in various techniques. Bioz Stars score: 88/100, 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.
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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.
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    Austral Biologicals mouse monoclonal α caga antibody
    Role of EPIYA motifs in <t>CagA</t> phosphorylation during H. pylori infection was investigated with seven different α-phosphotyrosine antibodies. AGS cells were infected for 6-expressing H. pylori strains as indicated. The samples in Figure 4 were harvested after photographing. Phosphorylation of CagA was examined using the indicated α–phosphotyrosine antibodies. Loading of equal amounts of CagA from each sample was confirmed by probing with a monoclonal <t>α-CagA</t> antibody. A larger section of the ∼120−180 kDa range is shown and contains the phospho-CagA bands of different sizes (arrows) as well as a set of tyrosine-phosphorylated host cell proteins (red asterisks). The blue asterisk indicates a putative N-terminal fragment of CagA which sometimes appears on SDS-PAGE gels [23] .
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    Santa Cruz Biotechnology polyclonal anti caga antibody b 300
    Analysis of the <t>CagA</t> protein patterns by immunoblot analysis. (A) Lysates of H . pylori isolated from the tissue specimens of various gastric patients were prepared as described in the Materials and Methods. Aliquots of protein (20 μg) were separated by SDS-gel electrophoresis and CagA was detected by immunoblotting using <t>polyclonal</t> anti-CagA antibody b-300 (Santa Cruz). The H . pylori 60190 and 26695 cagA knockout mutant (26695ΔA) were included as a positive and negative control, respectively, for CagA expression. The numbers at the top identify the various H . pylori colonies, while those on the left indicate the molecular weight markers in kDa. It can be seen that there are four types of CagA fragmentation patterns. (B) The percentages of ABD subtypes based on the CagA protein fragmentation patterns across the 96 H . pylori cagA ABD colonies that were analyzed and clearly ABD-2 is the predominant type.
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    Abnova pylori caga antibody
    (A) Comparison of the overall protein profiles in crude cell extracts of wild-type H . pylori 26695 (red) and the Hp Δ glyA deletion strain (blue) as determined by capillary electrophoresis (Experion, Bio-Rad) (left) and by migration on SDS-PAGE (right). The absence of a discrete protein band (*) with a molecular mass around 145 kDa is observed in the ΔglyA strain. (B) Immunoblot of crude extracts of H . pylori strains: 26695 ( glyA + , <t>cagA</t> + ), Hp Δ glyA ( glyA - , cagA + ) and X47 ( glyA + , cagA - ). Immunodetection was performed with antibodies directed against CagA and <t>UreB</t> proteins from H . pylori . (C) PCR amplification of a 307 bp internal fragment of cagA performed with the primer pair SA82/SA83 ( Table 1 ). Amplification is observed in the wild-type 26695 and Hp Δ glyA deletion strains, but not in the X47 control strain.
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    Oravax Inc anti recombinant caga protein mouse polyclonal antibody
    (A) Comparison of the overall protein profiles in crude cell extracts of wild-type H . pylori 26695 (red) and the Hp Δ glyA deletion strain (blue) as determined by capillary electrophoresis (Experion, Bio-Rad) (left) and by migration on SDS-PAGE (right). The absence of a discrete protein band (*) with a molecular mass around 145 kDa is observed in the ΔglyA strain. (B) Immunoblot of crude extracts of H . pylori strains: 26695 ( glyA + , <t>cagA</t> + ), Hp Δ glyA ( glyA - , cagA + ) and X47 ( glyA + , cagA - ). Immunodetection was performed with antibodies directed against CagA and <t>UreB</t> proteins from H . pylori . (C) PCR amplification of a 307 bp internal fragment of cagA performed with the primer pair SA82/SA83 ( Table 1 ). Amplification is observed in the wild-type 26695 and Hp Δ glyA deletion strains, but not in the X47 control strain.
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    Austral Biologicals anti caga polyclonal antibody hpp 5003 9
    (A) Comparison of the overall protein profiles in crude cell extracts of wild-type H . pylori 26695 (red) and the Hp Δ glyA deletion strain (blue) as determined by capillary electrophoresis (Experion, Bio-Rad) (left) and by migration on SDS-PAGE (right). The absence of a discrete protein band (*) with a molecular mass around 145 kDa is observed in the ΔglyA strain. (B) Immunoblot of crude extracts of H . pylori strains: 26695 ( glyA + , <t>cagA</t> + ), Hp Δ glyA ( glyA - , cagA + ) and X47 ( glyA + , cagA - ). Immunodetection was performed with antibodies directed against CagA and <t>UreB</t> proteins from H . pylori . (C) PCR amplification of a 307 bp internal fragment of cagA performed with the primer pair SA82/SA83 ( Table 1 ). Amplification is observed in the wild-type 26695 and Hp Δ glyA deletion strains, but not in the X47 control strain.
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    Novartis anti caga rabbit polyclonal antibodies
    (A) Comparison of the overall protein profiles in crude cell extracts of wild-type H . pylori 26695 (red) and the Hp Δ glyA deletion strain (blue) as determined by capillary electrophoresis (Experion, Bio-Rad) (left) and by migration on SDS-PAGE (right). The absence of a discrete protein band (*) with a molecular mass around 145 kDa is observed in the ΔglyA strain. (B) Immunoblot of crude extracts of H . pylori strains: 26695 ( glyA + , <t>cagA</t> + ), Hp Δ glyA ( glyA - , cagA + ) and X47 ( glyA + , cagA - ). Immunodetection was performed with antibodies directed against CagA and <t>UreB</t> proteins from H . pylori . (C) PCR amplification of a 307 bp internal fragment of cagA performed with the primer pair SA82/SA83 ( Table 1 ). Amplification is observed in the wild-type 26695 and Hp Δ glyA deletion strains, but not in the X47 control strain.
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    Variation in the CagA amino acid sequence of East Asian-type and Western-type CagA. The target α-EAS sequences “AINRKIDRINKIASAGKG” in EPIYA segment D, based on the sequences of Japanese strains, are shown in frame. A sequence analysis revealed that the typical target sequences in Bhutanese strains was “(T/K)IN(G/R)KID(Q/R)(L/I)N(R/K)(T/I)ASA(G/N)KG,” where (X/Y) means X and Y as the two major amino acids. The sequences of the EPIYA-D segments in Bhutanese strains were highly variable compared to strains deposited in GenBank. In contrast, the East Asian-type and Western-type sequences of EPIYA-C segments in Bhutanese strains were largely identical, similar to the typical sequences of EPIYA-C segments deposited in GenBank. Reference strains used in Fig. 1A (strain name [accession number]) were 103a (AB110966.1), 105a (AB110967.1), 106a (AB110968.1), 108a (AB110969.1), 113b (AB110970.1), 120a (AB110971.1), 122b (AB110972.1), 125b (AB110973.1), 128a (AB110974.1), FJT77 (KF028580.1), 04-518 (AB267252.1), 03-166 (AB267253.1), 04-264 (AB267254.1), THP1477 (AB116744.1), 04-334 (AB267249.1), 03-292 (AB267250.1), 04-366 (AB267251.1), THP1260 (AB116742.1), M3 (AB116740.1), THP463 (AB116735.1), Korea23 (AB057044.1), Korea 12 (AB057043.1), K69 (FJ458129.1), Korea2-3 (AB057040.1), k266 (FJ458163.1), K265 (FJ458162.1), K264 (FJ458161.1), K261 (FJ458158.1), K260 (FJ458157.1), K259 (FJ458156.1). Reference strains used in Fig. 1C (strain name [accession number]) were India41 (AF222807.1), India99 (AF222809.1), OSC40A (EU089774.1), OSC42B (EU089775.1), PCR-156i (EU368669.1), PCR218vi (EU089766.1). RIGLD-OC149 (JX428784.1), SAN53 (EU089771.1), PD682 (EF450167.1), PD636 (EF450165.1), PD308 (EF450162.1), PD488 (EF450161.1), PD537 (EF450160.1), PD501 (EF450159.1), PD351 (EF450158.1), PD348 (EF450157.1), PD6481K (EF450153.1), 216G (GQ899171.1), 1407 (GU143415.1), HPI-14 (FJ849792.1), HPI-13 (FJ849791.1), HPI-11 (FJ849789.1), USA2791 (AB057099.1), Kazak3 (AB057098.1), USA35 (AB057095.1), Italy329 (AB057094.1), Arizona2 (AB057075.1), Arizona1 (AB057074.1).

    Journal: Scientific Reports

    Article Title: Rare Helicobacter pylori Virulence Genotypes in Bhutan

    doi: 10.1038/srep22584

    Figure Lengend Snippet: Variation in the CagA amino acid sequence of East Asian-type and Western-type CagA. The target α-EAS sequences “AINRKIDRINKIASAGKG” in EPIYA segment D, based on the sequences of Japanese strains, are shown in frame. A sequence analysis revealed that the typical target sequences in Bhutanese strains was “(T/K)IN(G/R)KID(Q/R)(L/I)N(R/K)(T/I)ASA(G/N)KG,” where (X/Y) means X and Y as the two major amino acids. The sequences of the EPIYA-D segments in Bhutanese strains were highly variable compared to strains deposited in GenBank. In contrast, the East Asian-type and Western-type sequences of EPIYA-C segments in Bhutanese strains were largely identical, similar to the typical sequences of EPIYA-C segments deposited in GenBank. Reference strains used in Fig. 1A (strain name [accession number]) were 103a (AB110966.1), 105a (AB110967.1), 106a (AB110968.1), 108a (AB110969.1), 113b (AB110970.1), 120a (AB110971.1), 122b (AB110972.1), 125b (AB110973.1), 128a (AB110974.1), FJT77 (KF028580.1), 04-518 (AB267252.1), 03-166 (AB267253.1), 04-264 (AB267254.1), THP1477 (AB116744.1), 04-334 (AB267249.1), 03-292 (AB267250.1), 04-366 (AB267251.1), THP1260 (AB116742.1), M3 (AB116740.1), THP463 (AB116735.1), Korea23 (AB057044.1), Korea 12 (AB057043.1), K69 (FJ458129.1), Korea2-3 (AB057040.1), k266 (FJ458163.1), K265 (FJ458162.1), K264 (FJ458161.1), K261 (FJ458158.1), K260 (FJ458157.1), K259 (FJ458156.1). Reference strains used in Fig. 1C (strain name [accession number]) were India41 (AF222807.1), India99 (AF222809.1), OSC40A (EU089774.1), OSC42B (EU089775.1), PCR-156i (EU368669.1), PCR218vi (EU089766.1). RIGLD-OC149 (JX428784.1), SAN53 (EU089771.1), PD682 (EF450167.1), PD636 (EF450165.1), PD308 (EF450162.1), PD488 (EF450161.1), PD537 (EF450160.1), PD501 (EF450159.1), PD351 (EF450158.1), PD348 (EF450157.1), PD6481K (EF450153.1), 216G (GQ899171.1), 1407 (GU143415.1), HPI-14 (FJ849792.1), HPI-13 (FJ849791.1), HPI-11 (FJ849789.1), USA2791 (AB057099.1), Kazak3 (AB057098.1), USA35 (AB057095.1), Italy329 (AB057094.1), Arizona2 (AB057075.1), Arizona1 (AB057074.1).

    Article Snippet: Briefly, after antigen retrieval and inactivation of endogenous peroxidase activity, tissue sections were incubated with α-H. pylori antibody (DAKO, Glostrup, Denmark), anti-CagA antibody (b-300; Santa Cruz Biotechnology, Santa Cruz, CA, USA) or α-EAS Ab diluted 1:2,000 with diluting solution (DAKO) overnight at 4 °C.

    Techniques: Sequencing, Western Blot, Polymerase Chain Reaction

    CagA injection by H. pylori cannot be achieved in infected HEK293 cell lines but in AGS gastric epithelial cells. Western blot analysis of (A) AGS, (B) HEK293-TLR2, (C) HEK293 and (D) HEK293-TLR5 cells infected with H. pylori wild-type strains P1, P12, P310 or 26695 for 6 hours. Phosphorylation of injected CagA was monitored using phosphotyrosine α-PY-99 and α-CagA antibodies. Red arrows indicate the position of phosphorylated CagA on the blot. Western blots for the house keeping gene GAPDH served as loading control.

    Journal: PLoS ONE

    Article Title: Induction of TLR-2 and TLR-5 Expression by Helicobacter pylori Switches cagPAI-Dependent Signalling Leading to the Secretion of IL-8 and TNF-?

    doi: 10.1371/journal.pone.0019614

    Figure Lengend Snippet: CagA injection by H. pylori cannot be achieved in infected HEK293 cell lines but in AGS gastric epithelial cells. Western blot analysis of (A) AGS, (B) HEK293-TLR2, (C) HEK293 and (D) HEK293-TLR5 cells infected with H. pylori wild-type strains P1, P12, P310 or 26695 for 6 hours. Phosphorylation of injected CagA was monitored using phosphotyrosine α-PY-99 and α-CagA antibodies. Red arrows indicate the position of phosphorylated CagA on the blot. Western blots for the house keeping gene GAPDH served as loading control.

    Article Snippet: Phosphorylated and non-phosphorylated CagA proteins were detected by incubation of the membranes with a mouse monoclonal α-phosphotyrosine antibody PY99 (Santa Cruz, USA) and a rabbit polyclonal α-CagA antibody (Austral Biologicals, USA).

    Techniques: Injection, Infection, Western Blot

    PC activation induced by VacA and CagA on THP-1 cells. Both VacA and CagA significantly ( P

    Journal: Infection and Immunity

    Article Title: Role of Activated Protein C in Helicobacter pylori-Associated Gastritis

    doi:

    Figure Lengend Snippet: PC activation induced by VacA and CagA on THP-1 cells. Both VacA and CagA significantly ( P

    Article Snippet: Recombinant VacA toxin, recombinant CagA from H. pylori , and polyclonal anti-VacA and anti-CagA antibodies were purchased from Austral Biologicals (San Ramon, Calif.).

    Techniques: Activation Assay

    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

    Ubiquitination of total proteins and proteasome activity in AGS cells co-cultured with H. pylori . (A) Representative immunoblot of total ubiquitinated protein from AGS cells infected with H. pylori (MOI = 300) for 8 h, in presence or absence of 2.5 μM Lactacystin. Tubulin is shown as loading control and CagA as infection control. (B) Quantification of proteasome activity in AGS cells infected with H. pylori wild type and its isogenic mutants cagA, vacA and, in presence of 2.5 μM Lactacystin for 8 h. Asterisk represents significant difference ( p

    Journal: Frontiers in Cellular and Infection Microbiology

    Article Title: KCTD5 and Ubiquitin Proteasome Signaling Are Required for Helicobacter pylori Adherence

    doi: 10.3389/fcimb.2017.00450

    Figure Lengend Snippet: Ubiquitination of total proteins and proteasome activity in AGS cells co-cultured with H. pylori . (A) Representative immunoblot of total ubiquitinated protein from AGS cells infected with H. pylori (MOI = 300) for 8 h, in presence or absence of 2.5 μM Lactacystin. Tubulin is shown as loading control and CagA as infection control. (B) Quantification of proteasome activity in AGS cells infected with H. pylori wild type and its isogenic mutants cagA, vacA and, in presence of 2.5 μM Lactacystin for 8 h. Asterisk represents significant difference ( p

    Article Snippet: Anti-Tubulin mouse monoclonal (Catalog number: T5168, Sigma-Aldrich, St Louis, Missouri, USA) and anti-CagA mouse monoclonal antibodies (Catalog number: sc-28368, Santa Cruz Biotechnology, Dallas, Texas, USA) were used as loading and H. pylori -infection controls, respectively.

    Techniques: Activity Assay, Cell Culture, Infection

    Analysis of the CagA protein patterns by immunoblot analysis. (A) Lysates of H . pylori isolated from the tissue specimens of various gastric patients were prepared as described in the Materials and Methods. Aliquots of protein (20 μg) were separated by SDS-gel electrophoresis and CagA was detected by immunoblotting using polyclonal anti-CagA antibody b-300 (Santa Cruz). The H . pylori 60190 and 26695 cagA knockout mutant (26695ΔA) were included as a positive and negative control, respectively, for CagA expression. The numbers at the top identify the various H . pylori colonies, while those on the left indicate the molecular weight markers in kDa. It can be seen that there are four types of CagA fragmentation patterns. (B) The percentages of ABD subtypes based on the CagA protein fragmentation patterns across the 96 H . pylori cagA ABD colonies that were analyzed and clearly ABD-2 is the predominant type.

    Journal: PLoS ONE

    Article Title: Fragmentation of CagA Reduces Hummingbird Phenotype Induction by Helicobactor pylori

    doi: 10.1371/journal.pone.0150061

    Figure Lengend Snippet: Analysis of the CagA protein patterns by immunoblot analysis. (A) Lysates of H . pylori isolated from the tissue specimens of various gastric patients were prepared as described in the Materials and Methods. Aliquots of protein (20 μg) were separated by SDS-gel electrophoresis and CagA was detected by immunoblotting using polyclonal anti-CagA antibody b-300 (Santa Cruz). The H . pylori 60190 and 26695 cagA knockout mutant (26695ΔA) were included as a positive and negative control, respectively, for CagA expression. The numbers at the top identify the various H . pylori colonies, while those on the left indicate the molecular weight markers in kDa. It can be seen that there are four types of CagA fragmentation patterns. (B) The percentages of ABD subtypes based on the CagA protein fragmentation patterns across the 96 H . pylori cagA ABD colonies that were analyzed and clearly ABD-2 is the predominant type.

    Article Snippet: Three μg anti-CagA polyclonal antibody (b-300 Santa Cruz Biotechnology) were added to the supernatant and incubated overnight at 4°C.

    Techniques: Isolation, SDS-Gel, Electrophoresis, Knock-Out, Mutagenesis, Negative Control, Expressing, Molecular Weight

    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

    Role of EPIYA motifs in CagA phosphorylation during H. pylori infection was investigated with seven different α-phosphotyrosine antibodies. AGS cells were infected for 6-expressing H. pylori strains as indicated. The samples in Figure 4 were harvested after photographing. Phosphorylation of CagA was examined using the indicated α–phosphotyrosine antibodies. Loading of equal amounts of CagA from each sample was confirmed by probing with a monoclonal α-CagA antibody. A larger section of the ∼120−180 kDa range is shown and contains the phospho-CagA bands of different sizes (arrows) as well as a set of tyrosine-phosphorylated host cell proteins (red asterisks). The blue asterisk indicates a putative N-terminal fragment of CagA which sometimes appears on SDS-PAGE gels [23] .

    Journal: PLoS ONE

    Article Title: Systematic Analysis of Phosphotyrosine Antibodies Recognizing Single Phosphorylated EPIYA-Motifs in CagA of Western-Type Helicobacter pylori Strains

    doi: 10.1371/journal.pone.0096488

    Figure Lengend Snippet: Role of EPIYA motifs in CagA phosphorylation during H. pylori infection was investigated with seven different α-phosphotyrosine antibodies. AGS cells were infected for 6-expressing H. pylori strains as indicated. The samples in Figure 4 were harvested after photographing. Phosphorylation of CagA was examined using the indicated α–phosphotyrosine antibodies. Loading of equal amounts of CagA from each sample was confirmed by probing with a monoclonal α-CagA antibody. A larger section of the ∼120−180 kDa range is shown and contains the phospho-CagA bands of different sizes (arrows) as well as a set of tyrosine-phosphorylated host cell proteins (red asterisks). The blue asterisk indicates a putative N-terminal fragment of CagA which sometimes appears on SDS-PAGE gels [23] .

    Article Snippet: Membranes were incubated with the seven α-phosphotyrosine antibodies ( ) or mouse monoclonal α-CagA antibody (Austral Biologicals, San Ramon, CA, USA) according to the instructions of the manufacturer.

    Techniques: Infection, Expressing, SDS Page

    Analysis of the CagA protein patterns by immunoblot analysis. (A) Lysates of H . pylori isolated from the tissue specimens of various gastric patients were prepared as described in the Materials and Methods. Aliquots of protein (20 μg) were separated by SDS-gel electrophoresis and CagA was detected by immunoblotting using polyclonal anti-CagA antibody b-300 (Santa Cruz). The H . pylori 60190 and 26695 cagA knockout mutant (26695ΔA) were included as a positive and negative control, respectively, for CagA expression. The numbers at the top identify the various H . pylori colonies, while those on the left indicate the molecular weight markers in kDa. It can be seen that there are four types of CagA fragmentation patterns. (B) The percentages of ABD subtypes based on the CagA protein fragmentation patterns across the 96 H . pylori cagA ABD colonies that were analyzed and clearly ABD-2 is the predominant type.

    Journal: PLoS ONE

    Article Title: Fragmentation of CagA Reduces Hummingbird Phenotype Induction by Helicobactor pylori

    doi: 10.1371/journal.pone.0150061

    Figure Lengend Snippet: Analysis of the CagA protein patterns by immunoblot analysis. (A) Lysates of H . pylori isolated from the tissue specimens of various gastric patients were prepared as described in the Materials and Methods. Aliquots of protein (20 μg) were separated by SDS-gel electrophoresis and CagA was detected by immunoblotting using polyclonal anti-CagA antibody b-300 (Santa Cruz). The H . pylori 60190 and 26695 cagA knockout mutant (26695ΔA) were included as a positive and negative control, respectively, for CagA expression. The numbers at the top identify the various H . pylori colonies, while those on the left indicate the molecular weight markers in kDa. It can be seen that there are four types of CagA fragmentation patterns. (B) The percentages of ABD subtypes based on the CagA protein fragmentation patterns across the 96 H . pylori cagA ABD colonies that were analyzed and clearly ABD-2 is the predominant type.

    Article Snippet: Proteins (20 μg) were separated by SDS-gel electrophoresis, amounts of CagA and phosphorylated-CagA were determined by immunoblotting using polyclonal anti-CagA antibody b-300 (Santa Cruz) and anti-phosphotyrosine antibody PY99 (Santa Cruz).

    Techniques: Isolation, SDS-Gel, Electrophoresis, Knock-Out, Mutagenesis, Negative Control, Expressing, Molecular Weight

    (A) Comparison of the overall protein profiles in crude cell extracts of wild-type H . pylori 26695 (red) and the Hp Δ glyA deletion strain (blue) as determined by capillary electrophoresis (Experion, Bio-Rad) (left) and by migration on SDS-PAGE (right). The absence of a discrete protein band (*) with a molecular mass around 145 kDa is observed in the ΔglyA strain. (B) Immunoblot of crude extracts of H . pylori strains: 26695 ( glyA + , cagA + ), Hp Δ glyA ( glyA - , cagA + ) and X47 ( glyA + , cagA - ). Immunodetection was performed with antibodies directed against CagA and UreB proteins from H . pylori . (C) PCR amplification of a 307 bp internal fragment of cagA performed with the primer pair SA82/SA83 ( Table 1 ). Amplification is observed in the wild-type 26695 and Hp Δ glyA deletion strains, but not in the X47 control strain.

    Journal: PLoS ONE

    Article Title: Structural and functional insight into serine hydroxymethyltransferase from Helicobacter pylori

    doi: 10.1371/journal.pone.0208850

    Figure Lengend Snippet: (A) Comparison of the overall protein profiles in crude cell extracts of wild-type H . pylori 26695 (red) and the Hp Δ glyA deletion strain (blue) as determined by capillary electrophoresis (Experion, Bio-Rad) (left) and by migration on SDS-PAGE (right). The absence of a discrete protein band (*) with a molecular mass around 145 kDa is observed in the ΔglyA strain. (B) Immunoblot of crude extracts of H . pylori strains: 26695 ( glyA + , cagA + ), Hp Δ glyA ( glyA - , cagA + ) and X47 ( glyA + , cagA - ). Immunodetection was performed with antibodies directed against CagA and UreB proteins from H . pylori . (C) PCR amplification of a 307 bp internal fragment of cagA performed with the primer pair SA82/SA83 ( Table 1 ). Amplification is observed in the wild-type 26695 and Hp Δ glyA deletion strains, but not in the X47 control strain.

    Article Snippet: Mouse monoclonal anti-polyhistidine antibody (Sigma-Aldrich), rabbit polyclonal anti-UreB antibody (Abcam), and mouse monoclonal anti-H . pylori CagA antibody (Abnova) were used as primary antibodies.

    Techniques: Electrophoresis, Migration, SDS Page, Immunodetection, Polymerase Chain Reaction, Amplification