anti-syntaxin1a Search Results


anti-syntaxin-1a  (GeneTex)
90
GeneTex anti-syntaxin-1a
Anti Syntaxin 1a, supplied by GeneTex, used in various techniques. Bioz Stars score: 90/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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a rabbit polyclonal antibody against syntaxin 16  (Synaptic Systems)
90
Synaptic Systems a rabbit polyclonal antibody against syntaxin 16
( A , B ) Gastric cell populations were cultured for 24 h in gland culture medium containing ranitidine, then fixed in 1% formaldehyde, permeabilized and stained with either ( A ) TRITC-conjugated phalloidin, mouse anti-GM130 ( cis -Golgi marker) antibodies followed by FITC-conjugated anti-mouse IgG and rabbit anti-syntaxin 16 (TGN marker) antibodies followed by Alexa Fluor® 647-conjugated to anti-rabbit IgG or ( B ) mouse anti-HKα antibodies followed by FITC-conjugated anti-mouse IgG, rabbit antiGCC185 (TGN marker) antibodies followed by Alexa Fluor® 647-conjugated anti-rabbit IgG (pseudo-coloured green) and human anti-p230 (TGN marker) antibodies followed by Alexa Fluor® 594-conjugated to anti-human IgG. ( C ) Gastric glands were isolated from the stomachs of wild-type mice (fasted) and fixed in 4% PFA, permeabilized and stained with TRITC-conjugated phalloidin, mouse anti-GM130 antibodies followed by FITC-conjugated anti-mouse IgG and rabbit anti-syntaxin16 (TGN marker) antibodies followed by Alexa Fluor® 647-conjugated to anti-rabbit IgG. Scale bars represent 10 μm.
A Rabbit Polyclonal Antibody Against Syntaxin 16, supplied by Synaptic Systems, used in various techniques. Bioz Stars score: 90/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
https://www.bioz.com/result/a rabbit polyclonal antibody against syntaxin 16/product/Synaptic Systems
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a rabbit polyclonal antibody against syntaxin 16 - by Bioz Stars, 2026-06
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rabbit anti-syntaxin 13  (Synaptic Systems)
90
Synaptic Systems rabbit anti-syntaxin 13
Immunogold EM of hippocampal neurons labeled with 10 nm protein A gold for Rab4 and with 15 nm protein A gold for GRASP-1 (A), with 10 nm protein A gold for <t>syntaxin</t> <t>13</t> and with 15 nm protein A gold for GRASP-1 (B), with 10 nm protein A gold for syntaxin 13 and with 15 nm protein A gold for Rab4 (C), or with 15 nm protein A gold for GRASP-1, with 5 nm protein gold for syntaxin 13, and with 10 nm protein A gold for rab4 (D). Arrow denotes tubular endosomal membrane to which GRASP-1, syntaxin 13, and Rab4 localized. EE indicates early endosomes and scale bar is 100 nm.
Rabbit Anti Syntaxin 13, supplied by Synaptic Systems, used in various techniques. Bioz Stars score: 90/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
https://www.bioz.com/result/rabbit anti-syntaxin 13/product/Synaptic Systems
Average 90 stars, based on 1 article reviews
rabbit anti-syntaxin 13 - by Bioz Stars, 2026-06
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mouse anti-syntaxin-1a monoclonal antibody  (FUJIFILM)
90
FUJIFILM mouse anti-syntaxin-1a monoclonal antibody
Immunogold EM of hippocampal neurons labeled with 10 nm protein A gold for Rab4 and with 15 nm protein A gold for GRASP-1 (A), with 10 nm protein A gold for <t>syntaxin</t> <t>13</t> and with 15 nm protein A gold for GRASP-1 (B), with 10 nm protein A gold for syntaxin 13 and with 15 nm protein A gold for Rab4 (C), or with 15 nm protein A gold for GRASP-1, with 5 nm protein gold for syntaxin 13, and with 10 nm protein A gold for rab4 (D). Arrow denotes tubular endosomal membrane to which GRASP-1, syntaxin 13, and Rab4 localized. EE indicates early endosomes and scale bar is 100 nm.
Mouse Anti Syntaxin 1a Monoclonal Antibody, supplied by FUJIFILM, used in various techniques. Bioz Stars score: 90/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
https://www.bioz.com/result/mouse anti-syntaxin-1a monoclonal antibody/product/FUJIFILM
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mouse anti-syntaxin-1a monoclonal antibody - by Bioz Stars, 2026-06
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mouse monoclonal 110 111 (clone 78.3) anti-syntaxin-1a  (Synaptic Systems)
90
Synaptic Systems mouse monoclonal 110 111 (clone 78.3) anti-syntaxin-1a
Immunogold EM of hippocampal neurons labeled with 10 nm protein A gold for Rab4 and with 15 nm protein A gold for GRASP-1 (A), with 10 nm protein A gold for <t>syntaxin</t> <t>13</t> and with 15 nm protein A gold for GRASP-1 (B), with 10 nm protein A gold for syntaxin 13 and with 15 nm protein A gold for Rab4 (C), or with 15 nm protein A gold for GRASP-1, with 5 nm protein gold for syntaxin 13, and with 10 nm protein A gold for rab4 (D). Arrow denotes tubular endosomal membrane to which GRASP-1, syntaxin 13, and Rab4 localized. EE indicates early endosomes and scale bar is 100 nm.
Mouse Monoclonal 110 111 (Clone 78.3) Anti Syntaxin 1a, supplied by Synaptic Systems, used in various techniques. Bioz Stars score: 90/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
https://www.bioz.com/result/mouse monoclonal 110 111 (clone 78.3) anti-syntaxin-1a/product/Synaptic Systems
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anti-syntaxin 1a b 78.2, ascites  (Synaptic Systems)
90
Synaptic Systems anti-syntaxin 1a b 78.2, ascites
( A ) A rat brain preparation enriched in synaptosomes was incubated for 30 min at 37°C with increasing concentrations of His 6 -BoNT/B-LC (left) of His 6 -TAT-BoNT/B-LC (right) as described in the Methods section. Reactions were terminated by addition of SDS sample buffer and the amount of intact synaptobrevin-2 was analyzed by Western blot using the 69.1 antibody as probe. The arrow indicates the electrophoretic mobility of synaptobrevin-2 (Syb). <t>Anti-syntaxin</t> 1 (Stx1) blot shows equal loading of synaptosomal proteins. Shown are blots representative of three repetitions. ( B ) Quantification was expressed as the ratio of the signal intensities of synaptobrevin-2 relative to syntaxin1 when synaptosomes were incubated with His 6 -TAT-BoNT/B-LC (squares) or His 6 -BoNT/B-LC (circles). ( C ) 100 nM His 6 -BoNT/B-LC and His 6 -TAT-BoNT/B-LC (black bar) were introduced into SLO-permeabilized sperm cells by incubating at 37°C for 15 min. The AR was induced with 0.5 mM CaCl 2 and incubating as before. Sperm cells were fixed and the AR was measured by FITC-PSA binding as described in . Gray bars represent controls: background (control), AR stimulated by 0.5 mM CaCl 2 (Ca), lack of effect of the toxins on the basal AR (BoNT/B, TAT-BoNT/B) and inhibitory effect of the not-membrane penetrating version on the AR elicited by CaCl 2 . The data represent the mean ± SEM of at least three independent experiments. Data were evaluated before normalization with the program GraphPad Prism 8 using the one way Anova, Dunnett's multiple comparisons test. Different letters indicate statistical significance ( P <0.05).
Anti Syntaxin 1a B 78.2, Ascites, supplied by Synaptic Systems, used in various techniques. Bioz Stars score: 90/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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mouse monoclonal antibodies that recognize syntaxin-1  (Synaptic Systems)
90
Synaptic Systems mouse monoclonal antibodies that recognize syntaxin-1
( A ) A rat brain preparation enriched in synaptosomes was incubated for 30 min at 37°C with increasing concentrations of His 6 -BoNT/B-LC (left) of His 6 -TAT-BoNT/B-LC (right) as described in the Methods section. Reactions were terminated by addition of SDS sample buffer and the amount of intact synaptobrevin-2 was analyzed by Western blot using the 69.1 antibody as probe. The arrow indicates the electrophoretic mobility of synaptobrevin-2 (Syb). <t>Anti-syntaxin</t> 1 (Stx1) blot shows equal loading of synaptosomal proteins. Shown are blots representative of three repetitions. ( B ) Quantification was expressed as the ratio of the signal intensities of synaptobrevin-2 relative to syntaxin1 when synaptosomes were incubated with His 6 -TAT-BoNT/B-LC (squares) or His 6 -BoNT/B-LC (circles). ( C ) 100 nM His 6 -BoNT/B-LC and His 6 -TAT-BoNT/B-LC (black bar) were introduced into SLO-permeabilized sperm cells by incubating at 37°C for 15 min. The AR was induced with 0.5 mM CaCl 2 and incubating as before. Sperm cells were fixed and the AR was measured by FITC-PSA binding as described in . Gray bars represent controls: background (control), AR stimulated by 0.5 mM CaCl 2 (Ca), lack of effect of the toxins on the basal AR (BoNT/B, TAT-BoNT/B) and inhibitory effect of the not-membrane penetrating version on the AR elicited by CaCl 2 . The data represent the mean ± SEM of at least three independent experiments. Data were evaluated before normalization with the program GraphPad Prism 8 using the one way Anova, Dunnett's multiple comparisons test. Different letters indicate statistical significance ( P <0.05).
Mouse Monoclonal Antibodies That Recognize Syntaxin 1, supplied by Synaptic Systems, used in various techniques. Bioz Stars score: 90/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
https://www.bioz.com/result/mouse monoclonal antibodies that recognize syntaxin-1/product/Synaptic Systems
Average 90 stars, based on 1 article reviews
mouse monoclonal antibodies that recognize syntaxin-1 - by Bioz Stars, 2026-06
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mouse monoclonal 110 111 78.3) anti-syntaxin-1a  (Synaptic Systems)
90
Synaptic Systems mouse monoclonal 110 111 78.3) anti-syntaxin-1a
( A ) A rat brain preparation enriched in synaptosomes was incubated for 30 min at 37°C with increasing concentrations of His 6 -BoNT/B-LC (left) of His 6 -TAT-BoNT/B-LC (right) as described in the Methods section. Reactions were terminated by addition of SDS sample buffer and the amount of intact synaptobrevin-2 was analyzed by Western blot using the 69.1 antibody as probe. The arrow indicates the electrophoretic mobility of synaptobrevin-2 (Syb). <t>Anti-syntaxin</t> 1 (Stx1) blot shows equal loading of synaptosomal proteins. Shown are blots representative of three repetitions. ( B ) Quantification was expressed as the ratio of the signal intensities of synaptobrevin-2 relative to syntaxin1 when synaptosomes were incubated with His 6 -TAT-BoNT/B-LC (squares) or His 6 -BoNT/B-LC (circles). ( C ) 100 nM His 6 -BoNT/B-LC and His 6 -TAT-BoNT/B-LC (black bar) were introduced into SLO-permeabilized sperm cells by incubating at 37°C for 15 min. The AR was induced with 0.5 mM CaCl 2 and incubating as before. Sperm cells were fixed and the AR was measured by FITC-PSA binding as described in . Gray bars represent controls: background (control), AR stimulated by 0.5 mM CaCl 2 (Ca), lack of effect of the toxins on the basal AR (BoNT/B, TAT-BoNT/B) and inhibitory effect of the not-membrane penetrating version on the AR elicited by CaCl 2 . The data represent the mean ± SEM of at least three independent experiments. Data were evaluated before normalization with the program GraphPad Prism 8 using the one way Anova, Dunnett's multiple comparisons test. Different letters indicate statistical significance ( P <0.05).
Mouse Monoclonal 110 111 78.3) Anti Syntaxin 1a, supplied by Synaptic Systems, used in various techniques. Bioz Stars score: 90/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
https://www.bioz.com/result/mouse monoclonal 110 111 78.3) anti-syntaxin-1a/product/Synaptic Systems
Average 90 stars, based on 1 article reviews
mouse monoclonal 110 111 78.3) anti-syntaxin-1a - by Bioz Stars, 2026-06
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anti-syntaxin 1a monoclonal antibody  (FUJIFILM)
90
FUJIFILM anti-syntaxin 1a monoclonal antibody
( A ) A rat brain preparation enriched in synaptosomes was incubated for 30 min at 37°C with increasing concentrations of His 6 -BoNT/B-LC (left) of His 6 -TAT-BoNT/B-LC (right) as described in the Methods section. Reactions were terminated by addition of SDS sample buffer and the amount of intact synaptobrevin-2 was analyzed by Western blot using the 69.1 antibody as probe. The arrow indicates the electrophoretic mobility of synaptobrevin-2 (Syb). <t>Anti-syntaxin</t> 1 (Stx1) blot shows equal loading of synaptosomal proteins. Shown are blots representative of three repetitions. ( B ) Quantification was expressed as the ratio of the signal intensities of synaptobrevin-2 relative to syntaxin1 when synaptosomes were incubated with His 6 -TAT-BoNT/B-LC (squares) or His 6 -BoNT/B-LC (circles). ( C ) 100 nM His 6 -BoNT/B-LC and His 6 -TAT-BoNT/B-LC (black bar) were introduced into SLO-permeabilized sperm cells by incubating at 37°C for 15 min. The AR was induced with 0.5 mM CaCl 2 and incubating as before. Sperm cells were fixed and the AR was measured by FITC-PSA binding as described in . Gray bars represent controls: background (control), AR stimulated by 0.5 mM CaCl 2 (Ca), lack of effect of the toxins on the basal AR (BoNT/B, TAT-BoNT/B) and inhibitory effect of the not-membrane penetrating version on the AR elicited by CaCl 2 . The data represent the mean ± SEM of at least three independent experiments. Data were evaluated before normalization with the program GraphPad Prism 8 using the one way Anova, Dunnett's multiple comparisons test. Different letters indicate statistical significance ( P <0.05).
Anti Syntaxin 1a Monoclonal Antibody, supplied by FUJIFILM, used in various techniques. Bioz Stars score: 90/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
https://www.bioz.com/result/anti-syntaxin 1a monoclonal antibody/product/FUJIFILM
Average 90 stars, based on 1 article reviews
anti-syntaxin 1a monoclonal antibody - by Bioz Stars, 2026-06
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rabbit monoclonal anti-syntaxin-1a  (ABclonal Biotechnology)
90
ABclonal Biotechnology rabbit monoclonal anti-syntaxin-1a
( A ) A rat brain preparation enriched in synaptosomes was incubated for 30 min at 37°C with increasing concentrations of His 6 -BoNT/B-LC (left) of His 6 -TAT-BoNT/B-LC (right) as described in the Methods section. Reactions were terminated by addition of SDS sample buffer and the amount of intact synaptobrevin-2 was analyzed by Western blot using the 69.1 antibody as probe. The arrow indicates the electrophoretic mobility of synaptobrevin-2 (Syb). <t>Anti-syntaxin</t> 1 (Stx1) blot shows equal loading of synaptosomal proteins. Shown are blots representative of three repetitions. ( B ) Quantification was expressed as the ratio of the signal intensities of synaptobrevin-2 relative to syntaxin1 when synaptosomes were incubated with His 6 -TAT-BoNT/B-LC (squares) or His 6 -BoNT/B-LC (circles). ( C ) 100 nM His 6 -BoNT/B-LC and His 6 -TAT-BoNT/B-LC (black bar) were introduced into SLO-permeabilized sperm cells by incubating at 37°C for 15 min. The AR was induced with 0.5 mM CaCl 2 and incubating as before. Sperm cells were fixed and the AR was measured by FITC-PSA binding as described in . Gray bars represent controls: background (control), AR stimulated by 0.5 mM CaCl 2 (Ca), lack of effect of the toxins on the basal AR (BoNT/B, TAT-BoNT/B) and inhibitory effect of the not-membrane penetrating version on the AR elicited by CaCl 2 . The data represent the mean ± SEM of at least three independent experiments. Data were evaluated before normalization with the program GraphPad Prism 8 using the one way Anova, Dunnett's multiple comparisons test. Different letters indicate statistical significance ( P <0.05).
Rabbit Monoclonal Anti Syntaxin 1a, supplied by ABclonal Biotechnology, used in various techniques. Bioz Stars score: 90/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
https://www.bioz.com/result/rabbit monoclonal anti-syntaxin-1a/product/ABclonal Biotechnology
Average 90 stars, based on 1 article reviews
rabbit monoclonal anti-syntaxin-1a - by Bioz Stars, 2026-06
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rabbit anti-syntaxin1a  (GeneTex)
90
GeneTex rabbit anti-syntaxin1a
( A ) A rat brain preparation enriched in synaptosomes was incubated for 30 min at 37°C with increasing concentrations of His 6 -BoNT/B-LC (left) of His 6 -TAT-BoNT/B-LC (right) as described in the Methods section. Reactions were terminated by addition of SDS sample buffer and the amount of intact synaptobrevin-2 was analyzed by Western blot using the 69.1 antibody as probe. The arrow indicates the electrophoretic mobility of synaptobrevin-2 (Syb). <t>Anti-syntaxin</t> 1 (Stx1) blot shows equal loading of synaptosomal proteins. Shown are blots representative of three repetitions. ( B ) Quantification was expressed as the ratio of the signal intensities of synaptobrevin-2 relative to syntaxin1 when synaptosomes were incubated with His 6 -TAT-BoNT/B-LC (squares) or His 6 -BoNT/B-LC (circles). ( C ) 100 nM His 6 -BoNT/B-LC and His 6 -TAT-BoNT/B-LC (black bar) were introduced into SLO-permeabilized sperm cells by incubating at 37°C for 15 min. The AR was induced with 0.5 mM CaCl 2 and incubating as before. Sperm cells were fixed and the AR was measured by FITC-PSA binding as described in . Gray bars represent controls: background (control), AR stimulated by 0.5 mM CaCl 2 (Ca), lack of effect of the toxins on the basal AR (BoNT/B, TAT-BoNT/B) and inhibitory effect of the not-membrane penetrating version on the AR elicited by CaCl 2 . The data represent the mean ± SEM of at least three independent experiments. Data were evaluated before normalization with the program GraphPad Prism 8 using the one way Anova, Dunnett's multiple comparisons test. Different letters indicate statistical significance ( P <0.05).
Rabbit Anti Syntaxin1a, supplied by GeneTex, used in various techniques. Bioz Stars score: 90/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
https://www.bioz.com/result/rabbit anti-syntaxin1a/product/GeneTex
Average 90 stars, based on 1 article reviews
rabbit anti-syntaxin1a - by Bioz Stars, 2026-06
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rabbit polyclonal antisyntaxin1a sysy 110 302  (Synaptic Systems)
90
Synaptic Systems rabbit polyclonal antisyntaxin1a sysy 110 302
( A ) A rat brain preparation enriched in synaptosomes was incubated for 30 min at 37°C with increasing concentrations of His 6 -BoNT/B-LC (left) of His 6 -TAT-BoNT/B-LC (right) as described in the Methods section. Reactions were terminated by addition of SDS sample buffer and the amount of intact synaptobrevin-2 was analyzed by Western blot using the 69.1 antibody as probe. The arrow indicates the electrophoretic mobility of synaptobrevin-2 (Syb). <t>Anti-syntaxin</t> 1 (Stx1) blot shows equal loading of synaptosomal proteins. Shown are blots representative of three repetitions. ( B ) Quantification was expressed as the ratio of the signal intensities of synaptobrevin-2 relative to syntaxin1 when synaptosomes were incubated with His 6 -TAT-BoNT/B-LC (squares) or His 6 -BoNT/B-LC (circles). ( C ) 100 nM His 6 -BoNT/B-LC and His 6 -TAT-BoNT/B-LC (black bar) were introduced into SLO-permeabilized sperm cells by incubating at 37°C for 15 min. The AR was induced with 0.5 mM CaCl 2 and incubating as before. Sperm cells were fixed and the AR was measured by FITC-PSA binding as described in . Gray bars represent controls: background (control), AR stimulated by 0.5 mM CaCl 2 (Ca), lack of effect of the toxins on the basal AR (BoNT/B, TAT-BoNT/B) and inhibitory effect of the not-membrane penetrating version on the AR elicited by CaCl 2 . The data represent the mean ± SEM of at least three independent experiments. Data were evaluated before normalization with the program GraphPad Prism 8 using the one way Anova, Dunnett's multiple comparisons test. Different letters indicate statistical significance ( P <0.05).
Rabbit Polyclonal Antisyntaxin1a Sysy 110 302, supplied by Synaptic Systems, used in various techniques. Bioz Stars score: 90/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
https://www.bioz.com/result/rabbit polyclonal antisyntaxin1a sysy 110 302/product/Synaptic Systems
Average 90 stars, based on 1 article reviews
rabbit polyclonal antisyntaxin1a sysy 110 302 - by Bioz Stars, 2026-06
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Image Search Results


( A , B ) Gastric cell populations were cultured for 24 h in gland culture medium containing ranitidine, then fixed in 1% formaldehyde, permeabilized and stained with either ( A ) TRITC-conjugated phalloidin, mouse anti-GM130 ( cis -Golgi marker) antibodies followed by FITC-conjugated anti-mouse IgG and rabbit anti-syntaxin 16 (TGN marker) antibodies followed by Alexa Fluor® 647-conjugated to anti-rabbit IgG or ( B ) mouse anti-HKα antibodies followed by FITC-conjugated anti-mouse IgG, rabbit antiGCC185 (TGN marker) antibodies followed by Alexa Fluor® 647-conjugated anti-rabbit IgG (pseudo-coloured green) and human anti-p230 (TGN marker) antibodies followed by Alexa Fluor® 594-conjugated to anti-human IgG. ( C ) Gastric glands were isolated from the stomachs of wild-type mice (fasted) and fixed in 4% PFA, permeabilized and stained with TRITC-conjugated phalloidin, mouse anti-GM130 antibodies followed by FITC-conjugated anti-mouse IgG and rabbit anti-syntaxin16 (TGN marker) antibodies followed by Alexa Fluor® 647-conjugated to anti-rabbit IgG. Scale bars represent 10 μm.

Journal: Biology of the Cell

Article Title: The Golgi apparatus in the endomembrane-rich gastric parietal cells exist as functional stable mini-stacks dispersed throughout the cytoplasm

doi: 10.1042/BC20110074

Figure Lengend Snippet: ( A , B ) Gastric cell populations were cultured for 24 h in gland culture medium containing ranitidine, then fixed in 1% formaldehyde, permeabilized and stained with either ( A ) TRITC-conjugated phalloidin, mouse anti-GM130 ( cis -Golgi marker) antibodies followed by FITC-conjugated anti-mouse IgG and rabbit anti-syntaxin 16 (TGN marker) antibodies followed by Alexa Fluor® 647-conjugated to anti-rabbit IgG or ( B ) mouse anti-HKα antibodies followed by FITC-conjugated anti-mouse IgG, rabbit antiGCC185 (TGN marker) antibodies followed by Alexa Fluor® 647-conjugated anti-rabbit IgG (pseudo-coloured green) and human anti-p230 (TGN marker) antibodies followed by Alexa Fluor® 594-conjugated to anti-human IgG. ( C ) Gastric glands were isolated from the stomachs of wild-type mice (fasted) and fixed in 4% PFA, permeabilized and stained with TRITC-conjugated phalloidin, mouse anti-GM130 antibodies followed by FITC-conjugated anti-mouse IgG and rabbit anti-syntaxin16 (TGN marker) antibodies followed by Alexa Fluor® 647-conjugated to anti-rabbit IgG. Scale bars represent 10 μm.

Article Snippet: A rabbit polyclonal antibody against syntaxin 16 was purchased from Synaptic Systems (Gottingen, Germany).

Techniques: Cell Culture, Staining, Marker, Isolation

Immunogold EM of hippocampal neurons labeled with 10 nm protein A gold for Rab4 and with 15 nm protein A gold for GRASP-1 (A), with 10 nm protein A gold for syntaxin 13 and with 15 nm protein A gold for GRASP-1 (B), with 10 nm protein A gold for syntaxin 13 and with 15 nm protein A gold for Rab4 (C), or with 15 nm protein A gold for GRASP-1, with 5 nm protein gold for syntaxin 13, and with 10 nm protein A gold for rab4 (D). Arrow denotes tubular endosomal membrane to which GRASP-1, syntaxin 13, and Rab4 localized. EE indicates early endosomes and scale bar is 100 nm.

Journal: PLoS Biology

Article Title: Neuron Specific Rab4 Effector GRASP-1 Coordinates Membrane Specialization and Maturation of Recycling Endosomes

doi: 10.1371/journal.pbio.1000283

Figure Lengend Snippet: Immunogold EM of hippocampal neurons labeled with 10 nm protein A gold for Rab4 and with 15 nm protein A gold for GRASP-1 (A), with 10 nm protein A gold for syntaxin 13 and with 15 nm protein A gold for GRASP-1 (B), with 10 nm protein A gold for syntaxin 13 and with 15 nm protein A gold for Rab4 (C), or with 15 nm protein A gold for GRASP-1, with 5 nm protein gold for syntaxin 13, and with 10 nm protein A gold for rab4 (D). Arrow denotes tubular endosomal membrane to which GRASP-1, syntaxin 13, and Rab4 localized. EE indicates early endosomes and scale bar is 100 nm.

Article Snippet: The following antibodies were obtained from commercial sources: rabbit anti-GRASP-1 (AB96361), mouse anti-β-actin, mouse anti-GluR2 (Chemicon), mouse anti-Rab4, mouse anti-EEA1 (BD Biosciences), mouse anti-FLAG, mouse anti-MAP2, mouse anti-αtubulin (Sigma), mouse anti-GFP (Roche), mouse anti-bassoon (Stressgen), rabbit anti-β-galactosidase (MP Biomedicals), mouse anti-β-galactosidase (Promega), rabbit anti-GluR1 (Calbiochem), mouse anti-HA (Roche), rabbit anti-myc (Upstate Biotechnology), mouse LAMP-1 (Stressgen), mouse anti-myc, rabbit anti-Rab5 (Santa Cruz Biotechnology), rabbit anti-syntaxin 13 (Synaptic Systems), human anti-EEA1, and mouse anti-human TfR (ATCC).

Techniques: Labeling

(A) Lysates of COS-7 cells cotransfected with GFP-GRASP-1 and myc-syntaxins were immunoprecipitated with anti-GFP antibody and analyzed by Western blot. (B) Lysates of COS-7 cells cotransfected with GFP-syntaxin 13 and full-length myc-GRASP-1 (1–837) or truncated myc-GRASP-1 constructs (1–695 or 695–837) were immunoprecipitated with anti-GFP antibody and analyzed by Western blot. Asterisk indicates background band. Arrows point to co-precipitated GRASP-1 proteins. (C) Binding assay using lysates of COS-7 cells expressing myc-syntaxin 13 with or without GFP-GRASP-1 and GMP-PNP-charged GST-rab4. Note that myc-syntaxin 13 is only isolated on the beads in the presence of GRASP-1. (D) Binding assay using lysate of COS-7 cells transfected with GFP-GRASP-1(594–837) and GST-syntaxins without transmembrane domain (ΔTM). GRASP-1 was analyzed by Western blot with antibody against GFP. (E) Binding assay of 35 S-labeled GRASP-1 and immobilized GST-syntaxin 13ΔTM.

Journal: PLoS Biology

Article Title: Neuron Specific Rab4 Effector GRASP-1 Coordinates Membrane Specialization and Maturation of Recycling Endosomes

doi: 10.1371/journal.pbio.1000283

Figure Lengend Snippet: (A) Lysates of COS-7 cells cotransfected with GFP-GRASP-1 and myc-syntaxins were immunoprecipitated with anti-GFP antibody and analyzed by Western blot. (B) Lysates of COS-7 cells cotransfected with GFP-syntaxin 13 and full-length myc-GRASP-1 (1–837) or truncated myc-GRASP-1 constructs (1–695 or 695–837) were immunoprecipitated with anti-GFP antibody and analyzed by Western blot. Asterisk indicates background band. Arrows point to co-precipitated GRASP-1 proteins. (C) Binding assay using lysates of COS-7 cells expressing myc-syntaxin 13 with or without GFP-GRASP-1 and GMP-PNP-charged GST-rab4. Note that myc-syntaxin 13 is only isolated on the beads in the presence of GRASP-1. (D) Binding assay using lysate of COS-7 cells transfected with GFP-GRASP-1(594–837) and GST-syntaxins without transmembrane domain (ΔTM). GRASP-1 was analyzed by Western blot with antibody against GFP. (E) Binding assay of 35 S-labeled GRASP-1 and immobilized GST-syntaxin 13ΔTM.

Article Snippet: The following antibodies were obtained from commercial sources: rabbit anti-GRASP-1 (AB96361), mouse anti-β-actin, mouse anti-GluR2 (Chemicon), mouse anti-Rab4, mouse anti-EEA1 (BD Biosciences), mouse anti-FLAG, mouse anti-MAP2, mouse anti-αtubulin (Sigma), mouse anti-GFP (Roche), mouse anti-bassoon (Stressgen), rabbit anti-β-galactosidase (MP Biomedicals), mouse anti-β-galactosidase (Promega), rabbit anti-GluR1 (Calbiochem), mouse anti-HA (Roche), rabbit anti-myc (Upstate Biotechnology), mouse LAMP-1 (Stressgen), mouse anti-myc, rabbit anti-Rab5 (Santa Cruz Biotechnology), rabbit anti-syntaxin 13 (Synaptic Systems), human anti-EEA1, and mouse anti-human TfR (ATCC).

Techniques: Immunoprecipitation, Western Blot, Construct, Binding Assay, Expressing, Isolation, Transfection, Labeling

(A) Representative image of hippocampal neuron triple transfected at DIV13 for 4 d with GFP-Rab4, HA-GRASP-1, and myc-syntaxin 13 and labeled with anti-HA (blue) or anti-myc (red) antibodies. Magnified region of the cell body is shown to indicate the strong colocalization of GRASP-1, Rab4, and syntaxin 13. (B) Representative images of dendrites of hippocampal neurons transfected at DIV13 with GFP-GRASP-1 for 4 d and labeled with anti-syntaxin 13 (red). (C) Representative images of dendrites of hippocampal neurons transfected at DIV13 with GFP-GRASP-1 for 4 d and labeled with anti-Neep21 (red). (D) Representative images of dendrites of hippocampal neurons cotransfected at DIV13 for 4 d with myc-syntaxin 13 and control vector or HA-GRASP-1 and labeled with anti-myc (green), anti-HA (blue), and anti-Neep21 (red). (E) Representative images of dendrites of hippocampal neurons cotransfected at DIV13 for 4 d with GFP-Rab4, HA-Rab11, and control vector or myc-syntaxin 13ΔTM and labeled with anti-myc (blue) and anti-HA (red). (F) Percentage of colocalization between HA-GRASP-1 and myc-syntaxin 1 or myc-syntaxin 13 in neurons. (G) Percentage of colocalization between myc-syntaxin 13 and Neep21 in dendrites as indicated in (D). (H) Percentage of colocalization between GFP-Rab4 and HA-Rab11 domains in dendrites expressing myc-syntaxin 13ΔTM as indicated in (E). Error bars indicate S.E.M. ** p <0.005. *** p <0.0005. Bar in A is 10 µm; Bar in (B–E) is 1 µm.

Journal: PLoS Biology

Article Title: Neuron Specific Rab4 Effector GRASP-1 Coordinates Membrane Specialization and Maturation of Recycling Endosomes

doi: 10.1371/journal.pbio.1000283

Figure Lengend Snippet: (A) Representative image of hippocampal neuron triple transfected at DIV13 for 4 d with GFP-Rab4, HA-GRASP-1, and myc-syntaxin 13 and labeled with anti-HA (blue) or anti-myc (red) antibodies. Magnified region of the cell body is shown to indicate the strong colocalization of GRASP-1, Rab4, and syntaxin 13. (B) Representative images of dendrites of hippocampal neurons transfected at DIV13 with GFP-GRASP-1 for 4 d and labeled with anti-syntaxin 13 (red). (C) Representative images of dendrites of hippocampal neurons transfected at DIV13 with GFP-GRASP-1 for 4 d and labeled with anti-Neep21 (red). (D) Representative images of dendrites of hippocampal neurons cotransfected at DIV13 for 4 d with myc-syntaxin 13 and control vector or HA-GRASP-1 and labeled with anti-myc (green), anti-HA (blue), and anti-Neep21 (red). (E) Representative images of dendrites of hippocampal neurons cotransfected at DIV13 for 4 d with GFP-Rab4, HA-Rab11, and control vector or myc-syntaxin 13ΔTM and labeled with anti-myc (blue) and anti-HA (red). (F) Percentage of colocalization between HA-GRASP-1 and myc-syntaxin 1 or myc-syntaxin 13 in neurons. (G) Percentage of colocalization between myc-syntaxin 13 and Neep21 in dendrites as indicated in (D). (H) Percentage of colocalization between GFP-Rab4 and HA-Rab11 domains in dendrites expressing myc-syntaxin 13ΔTM as indicated in (E). Error bars indicate S.E.M. ** p <0.005. *** p <0.0005. Bar in A is 10 µm; Bar in (B–E) is 1 µm.

Article Snippet: The following antibodies were obtained from commercial sources: rabbit anti-GRASP-1 (AB96361), mouse anti-β-actin, mouse anti-GluR2 (Chemicon), mouse anti-Rab4, mouse anti-EEA1 (BD Biosciences), mouse anti-FLAG, mouse anti-MAP2, mouse anti-αtubulin (Sigma), mouse anti-GFP (Roche), mouse anti-bassoon (Stressgen), rabbit anti-β-galactosidase (MP Biomedicals), mouse anti-β-galactosidase (Promega), rabbit anti-GluR1 (Calbiochem), mouse anti-HA (Roche), rabbit anti-myc (Upstate Biotechnology), mouse LAMP-1 (Stressgen), mouse anti-myc, rabbit anti-Rab5 (Santa Cruz Biotechnology), rabbit anti-syntaxin 13 (Synaptic Systems), human anti-EEA1, and mouse anti-human TfR (ATCC).

Techniques: Transfection, Labeling, Plasmid Preparation, Expressing

Endosomes can be viewed as mosaic distribution of Rab4, Rab5, and Rab11 domains that dynamically interact via effector proteins and SNAREs. The Rab5 domain allows entry into the early/sorting endosome, whereas the Rab4 and Rab11 domains contain the machinery that is necessary for sorting and recycling membranes and receptors back to the plasma membrane. (A) GRASP-1 binds to Rab4 and syntaxin 13 and couples Rab4 and Rab11 recycling endosomes. The complex formed between GRASP-1 and t-SNARE syntaxin 13 might mediate fusion between Rab4 and Rab11 endosomes. (B) Absence of GRASP-1 interferes with complex formation at the recycling step, causing cargo accumulation in early endosomes, impairment of receptor expression, and changes in spine morphology. (C) Overexpression of GRASP-1 leads to recruitment of syntaxin 13 and strongly couples Rab4 and Rab11 domains, causing accumulation of internalized receptors in recycling endosomes. Consistent with the observed decrease in AMPAR clusters , Caspase-3 cleavage of GRASP-1 might separate the N-terminal Rab4 domain from the C-terminal syntaxin 13 binding site and disrupt the coupling between Rab4 and Rab11 domains.

Journal: PLoS Biology

Article Title: Neuron Specific Rab4 Effector GRASP-1 Coordinates Membrane Specialization and Maturation of Recycling Endosomes

doi: 10.1371/journal.pbio.1000283

Figure Lengend Snippet: Endosomes can be viewed as mosaic distribution of Rab4, Rab5, and Rab11 domains that dynamically interact via effector proteins and SNAREs. The Rab5 domain allows entry into the early/sorting endosome, whereas the Rab4 and Rab11 domains contain the machinery that is necessary for sorting and recycling membranes and receptors back to the plasma membrane. (A) GRASP-1 binds to Rab4 and syntaxin 13 and couples Rab4 and Rab11 recycling endosomes. The complex formed between GRASP-1 and t-SNARE syntaxin 13 might mediate fusion between Rab4 and Rab11 endosomes. (B) Absence of GRASP-1 interferes with complex formation at the recycling step, causing cargo accumulation in early endosomes, impairment of receptor expression, and changes in spine morphology. (C) Overexpression of GRASP-1 leads to recruitment of syntaxin 13 and strongly couples Rab4 and Rab11 domains, causing accumulation of internalized receptors in recycling endosomes. Consistent with the observed decrease in AMPAR clusters , Caspase-3 cleavage of GRASP-1 might separate the N-terminal Rab4 domain from the C-terminal syntaxin 13 binding site and disrupt the coupling between Rab4 and Rab11 domains.

Article Snippet: The following antibodies were obtained from commercial sources: rabbit anti-GRASP-1 (AB96361), mouse anti-β-actin, mouse anti-GluR2 (Chemicon), mouse anti-Rab4, mouse anti-EEA1 (BD Biosciences), mouse anti-FLAG, mouse anti-MAP2, mouse anti-αtubulin (Sigma), mouse anti-GFP (Roche), mouse anti-bassoon (Stressgen), rabbit anti-β-galactosidase (MP Biomedicals), mouse anti-β-galactosidase (Promega), rabbit anti-GluR1 (Calbiochem), mouse anti-HA (Roche), rabbit anti-myc (Upstate Biotechnology), mouse LAMP-1 (Stressgen), mouse anti-myc, rabbit anti-Rab5 (Santa Cruz Biotechnology), rabbit anti-syntaxin 13 (Synaptic Systems), human anti-EEA1, and mouse anti-human TfR (ATCC).

Techniques: Expressing, Over Expression, Binding Assay

( A ) A rat brain preparation enriched in synaptosomes was incubated for 30 min at 37°C with increasing concentrations of His 6 -BoNT/B-LC (left) of His 6 -TAT-BoNT/B-LC (right) as described in the Methods section. Reactions were terminated by addition of SDS sample buffer and the amount of intact synaptobrevin-2 was analyzed by Western blot using the 69.1 antibody as probe. The arrow indicates the electrophoretic mobility of synaptobrevin-2 (Syb). Anti-syntaxin 1 (Stx1) blot shows equal loading of synaptosomal proteins. Shown are blots representative of three repetitions. ( B ) Quantification was expressed as the ratio of the signal intensities of synaptobrevin-2 relative to syntaxin1 when synaptosomes were incubated with His 6 -TAT-BoNT/B-LC (squares) or His 6 -BoNT/B-LC (circles). ( C ) 100 nM His 6 -BoNT/B-LC and His 6 -TAT-BoNT/B-LC (black bar) were introduced into SLO-permeabilized sperm cells by incubating at 37°C for 15 min. The AR was induced with 0.5 mM CaCl 2 and incubating as before. Sperm cells were fixed and the AR was measured by FITC-PSA binding as described in . Gray bars represent controls: background (control), AR stimulated by 0.5 mM CaCl 2 (Ca), lack of effect of the toxins on the basal AR (BoNT/B, TAT-BoNT/B) and inhibitory effect of the not-membrane penetrating version on the AR elicited by CaCl 2 . The data represent the mean ± SEM of at least three independent experiments. Data were evaluated before normalization with the program GraphPad Prism 8 using the one way Anova, Dunnett's multiple comparisons test. Different letters indicate statistical significance ( P <0.05).

Journal: Bioscience Reports

Article Title: Expression, purification and application of a recombinant, membrane permeating version of the light chain of botulinum toxin B

doi: 10.1042/BSR20240117

Figure Lengend Snippet: ( A ) A rat brain preparation enriched in synaptosomes was incubated for 30 min at 37°C with increasing concentrations of His 6 -BoNT/B-LC (left) of His 6 -TAT-BoNT/B-LC (right) as described in the Methods section. Reactions were terminated by addition of SDS sample buffer and the amount of intact synaptobrevin-2 was analyzed by Western blot using the 69.1 antibody as probe. The arrow indicates the electrophoretic mobility of synaptobrevin-2 (Syb). Anti-syntaxin 1 (Stx1) blot shows equal loading of synaptosomal proteins. Shown are blots representative of three repetitions. ( B ) Quantification was expressed as the ratio of the signal intensities of synaptobrevin-2 relative to syntaxin1 when synaptosomes were incubated with His 6 -TAT-BoNT/B-LC (squares) or His 6 -BoNT/B-LC (circles). ( C ) 100 nM His 6 -BoNT/B-LC and His 6 -TAT-BoNT/B-LC (black bar) were introduced into SLO-permeabilized sperm cells by incubating at 37°C for 15 min. The AR was induced with 0.5 mM CaCl 2 and incubating as before. Sperm cells were fixed and the AR was measured by FITC-PSA binding as described in . Gray bars represent controls: background (control), AR stimulated by 0.5 mM CaCl 2 (Ca), lack of effect of the toxins on the basal AR (BoNT/B, TAT-BoNT/B) and inhibitory effect of the not-membrane penetrating version on the AR elicited by CaCl 2 . The data represent the mean ± SEM of at least three independent experiments. Data were evaluated before normalization with the program GraphPad Prism 8 using the one way Anova, Dunnett's multiple comparisons test. Different letters indicate statistical significance ( P <0.05).

Article Snippet: The mouse monoclonals anti-synaptobrevin-2 antibody (clone 69.1, purified IgG) and anti-syntaxin 1A and B (clone 78.2, ascites) were from Synaptic Systems (Göttingen, Germany).

Techniques: Incubation, Western Blot, Binding Assay, Control, Membrane