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anti human arc antibody (Proteintech)

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Proteintech anti human arc antibody
Anti Human Arc Antibody, supplied by Proteintech, used in various techniques. Bioz Stars score: 93/100, based on 24 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
https://www.bioz.com/product/anti+human+arc+antibody/ARC+Antibody/pmc12739127-157-7-20
Average 93 stars, based on 24 article reviews

anti human arc antibody - by Bioz Stars, 2026-10

93/100 stars

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Incubation:

Article Title: The inner ear is a barometric pressure sensor-change in barometric pressure induces vestibular ganglion cell activation in mice.
Article Snippet: They were then washed with PBS for 20 min, with one buffer change, and rinsed with 0.3% Triton X-100 in 0.1 M PBS (PBST, pH 7.4) for 20 min, with one buffer change; nonspecific binding sites were blocked by incubating the sections in 25% Block Ace (DS Pharma Biomedical, Osaka, Japan) in PBST for 20 min at room temperature. .. The sections were subsequently incubated with an anti-human Arc antibody (rabbit polyclonal IgG: ARC/ARC3.1 Rabbit Polyclonal antibody; lot number 00094124; Proteintech, Rosemont, IL, USA) diluted 5,000 times with 10% Block Ace in PBST for approximately 40 h at 4°C. .. After three 10-min washes with PBST, the sections were incubated with a biotinylated anti-rabbit secondary antibody (BA-1000; Vector Laboratories, Burlingame, CA, USA) diluted 500 times with PBST for 2 h at room temperature and then processed with the ABC kit (VECTASTAIN Elite ABC kit PK-6100; Vector Laboratories, Burlingame, CA) appropriately diluted with PBST.

Article Title: The inner ear is a barometric pressure sensor—change in barometric pressure induces vestibular ganglion cell activation in mice
Article Snippet: They were then washed with PBS for 20 min, with one buffer change, and rinsed with 0.3% Triton X-100 in 0.1 M PBS (PBST, pH 7.4) for 20 min, with one buffer change; nonspecific binding sites were blocked by incubating the sections in 25% Block Ace (DS Pharma Biomedical, Osaka, Japan) in PBST for 20 min at room temperature. .. The sections were subsequently incubated with an anti-human Arc antibody (rabbit polyclonal IgG: ARC/ARC3.1 Rabbit Polyclonal antibody; lot number 00094124; Proteintech, Rosemont, IL, USA) diluted 5,000 times with 10% Block Ace in PBST for approximately 40 h at 4°C. .. After three 10-min washes with PBST, the sections were incubated with a biotinylated anti-rabbit secondary antibody (BA-1000; Vector Laboratories, Burlingame, CA, USA) diluted 500 times with PBST for 2 h at room temperature and then processed with the ABC kit (VECTASTAIN Elite ABC kit PK-6100; Vector Laboratories, Burlingame, CA) appropriately diluted with PBST.

Blocking Assay:

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( A ) PC12 cells were incubated for 2 min in low-K + solution. DA release during this period represented the basal release. The buffer was immediately changed every 2 min with the low-K + solution with or without 1 μM ionomycin to stimulate Ca 2+ influx. Samples were collected and tested according to the release assay in Materials and Methods. The amount of DA release in the medium was expressed as the percentage of total cellular content (n = 8/sample). ( B ) Cerebellar granule cells were performed similar to PC12 cells, and the amount of Glu released in the medium was expressed as the percentage of total cellular content (n = 8/sample). ( C ) PC12 cells were incubated with or without 1 μM ionomycin, lysed, and subjected to western blot using an anti-phosphotyrosine (P-Tyr, 4G10) antibody. Arrows indicate the major immunoreactive bands co-migrating with Pyk2 (upper) and paxillin (lower). ( D ) PC12 cell lysates were subjected to IP with anti-Pyk2 antibody, followed by IB with anti-phosphotyrosine and anti-Pyk2 antibodies. ( E ) PC12 cells were incubated for 2 min at 37°C in low-K + solution with or without 1 μM ionomycin. Equal amounts of cell lysates were immunoprecipitated with anti-Pyk2 antibody, and immunoblotted with phosphotyrosine antibody. Phosphorylation levels were studied in different experiments from 0 min to 5 min (right, inset) or from 1.5 min to 60 min (left, main graph). Pyk2 phosphorylation at each indicated time is expressed as a percentage of the maximal level of induced phosphorylation. Note: Duration of ionomycin treatment is indicated by the horizontal bar, which represents 2 min. The values in the results are expressed as mean ± S.E.M. from four representative experiments ( ## p<0.01, **p<0.01, ***p<0.001).

Journal: PLoS ONE

Article Title: Tyrosine 402 Phosphorylation of Pyk2 Is Involved in Ionomycin-Induced Neurotransmitter Release

doi: 10.1371/journal.pone.0094574

Figure Lengend Snippet: ( A ) PC12 cells were incubated for 2 min in low-K + solution. DA release during this period represented the basal release. The buffer was immediately changed every 2 min with the low-K + solution with or without 1 μM ionomycin to stimulate Ca 2+ influx. Samples were collected and tested according to the release assay in Materials and Methods. The amount of DA release in the medium was expressed as the percentage of total cellular content (n = 8/sample). ( B ) Cerebellar granule cells were performed similar to PC12 cells, and the amount of Glu released in the medium was expressed as the percentage of total cellular content (n = 8/sample). ( C ) PC12 cells were incubated with or without 1 μM ionomycin, lysed, and subjected to western blot using an anti-phosphotyrosine (P-Tyr, 4G10) antibody. Arrows indicate the major immunoreactive bands co-migrating with Pyk2 (upper) and paxillin (lower). ( D ) PC12 cell lysates were subjected to IP with anti-Pyk2 antibody, followed by IB with anti-phosphotyrosine and anti-Pyk2 antibodies. ( E ) PC12 cells were incubated for 2 min at 37°C in low-K + solution with or without 1 μM ionomycin. Equal amounts of cell lysates were immunoprecipitated with anti-Pyk2 antibody, and immunoblotted with phosphotyrosine antibody. Phosphorylation levels were studied in different experiments from 0 min to 5 min (right, inset) or from 1.5 min to 60 min (left, main graph). Pyk2 phosphorylation at each indicated time is expressed as a percentage of the maximal level of induced phosphorylation. Note: Duration of ionomycin treatment is indicated by the horizontal bar, which represents 2 min. The values in the results are expressed as mean ± S.E.M. from four representative experiments ( ## p<0.01, **p<0.01, ***p<0.001).

Article Snippet: The following antibodies were used: rabbit polyclonal Pyk2 phosphorylation site-specific antibody (pY402, pY579, pY580, and pY881) from Biosource International (Camarillo, CA, USA); anti-phosphotyrosine monoclonal antibody (clone 4G10) from Upstate Biotechnology (Lake Placid, NY, USA); mouse monoclonal anti-Pyk2, actin antibodies, rabbit polyclonal anti-FAK, Src, actin, Fyn antibodies, horseradish peroxidase (HRP)-conjugated anti-mouse IgG, anti-rabbit IgG, rhodamine (TRITC)-conjugated anti-rabbit IgG, FITC-conjugated anti-mouse IgG, and Human Pyk2 transfected 293T whole cell lysate (sc-115595) were obtained from Santa Cruz Biotechnology (Santa Cruz, CA, USA).

Techniques: Incubation, Release Assay, Western Blot, Immunoprecipitation, Phospho-proteomics

( A ) PC12 cells were transfected with 50 nM Pyk2 siRNA or control siRNA for 48 h, and tested for the expression of Pyk2 and FAK. PC12 cells were transfected with 50 nM Pyk2 siRNA for 24, 48, and 72 h or 50 nM control siRNA for 72 h, and harvested for western blot. PC12 cells were transfected with 10, 50, and 100 nM Pyk2 siRNA or 100 nM control siRNA for 48 h, and harvested for western blot. ( B ) PC12 cells were transfected with 100 nM Pyk2 siRNA or control siRNA for 48 h, and used for release assay with or without 1 μM ionomycin for 2 min. Samples were collected and tested for DA release. The amount of DA released in the medium was expressed as the percentage of total cellular content (n = 8/sample). The values are expressed as mean ± S.E.M. from four representative experiments (**p<0.01, ***p<0.001).

Journal: PLoS ONE

Article Title: Tyrosine 402 Phosphorylation of Pyk2 Is Involved in Ionomycin-Induced Neurotransmitter Release

doi: 10.1371/journal.pone.0094574

Figure Lengend Snippet: ( A ) PC12 cells were transfected with 50 nM Pyk2 siRNA or control siRNA for 48 h, and tested for the expression of Pyk2 and FAK. PC12 cells were transfected with 50 nM Pyk2 siRNA for 24, 48, and 72 h or 50 nM control siRNA for 72 h, and harvested for western blot. PC12 cells were transfected with 10, 50, and 100 nM Pyk2 siRNA or 100 nM control siRNA for 48 h, and harvested for western blot. ( B ) PC12 cells were transfected with 100 nM Pyk2 siRNA or control siRNA for 48 h, and used for release assay with or without 1 μM ionomycin for 2 min. Samples were collected and tested for DA release. The amount of DA released in the medium was expressed as the percentage of total cellular content (n = 8/sample). The values are expressed as mean ± S.E.M. from four representative experiments (**p<0.01, ***p<0.001).

Techniques: Transfection, Control, Expressing, Western Blot, Release Assay

( A ) PC12 cells were exposed to 1 μM ionomycin at 37°C for 90 s. The cells were fixed, and phosphotyrosine proteins were labeled by phosphorylation site-specific antibodies against Tyr-402 (pY402), Tyr-579 (pY579), Tyr-580 (pY580), or Tyr-881 (pY881) and TRITC-conjugated second antibody (red), respectively. Pyk2 expression was labeled by FITC (green). Scale bar, 5 μm. ( B ) PC12 cells were treated with or without 1 μM ionomycin at 37°C for 90 s, then lysed, and subjected to western blot using phosphorylation site-specific antibodies. Pyk2 transfected 293T cell lysate was used as a positive control. The total amount of Pyk2 was used as an internal control. ( C ) The time course of Pyk2 phosphorylation at Tyr-402 by ionomycin in PC12 cells. PC12 cells were exposed to 1 μM ionomycin at 37°C for the indicated times. The cells were labeled with pY402 site-specific polyclonal antibody, followed by a secondary layer of TRITC-conjugated second antibody to rabbit IgG (red). Scale bar, 10 μm.

Journal: PLoS ONE

Article Title: Tyrosine 402 Phosphorylation of Pyk2 Is Involved in Ionomycin-Induced Neurotransmitter Release

doi: 10.1371/journal.pone.0094574

Figure Lengend Snippet: ( A ) PC12 cells were exposed to 1 μM ionomycin at 37°C for 90 s. The cells were fixed, and phosphotyrosine proteins were labeled by phosphorylation site-specific antibodies against Tyr-402 (pY402), Tyr-579 (pY579), Tyr-580 (pY580), or Tyr-881 (pY881) and TRITC-conjugated second antibody (red), respectively. Pyk2 expression was labeled by FITC (green). Scale bar, 5 μm. ( B ) PC12 cells were treated with or without 1 μM ionomycin at 37°C for 90 s, then lysed, and subjected to western blot using phosphorylation site-specific antibodies. Pyk2 transfected 293T cell lysate was used as a positive control. The total amount of Pyk2 was used as an internal control. ( C ) The time course of Pyk2 phosphorylation at Tyr-402 by ionomycin in PC12 cells. PC12 cells were exposed to 1 μM ionomycin at 37°C for the indicated times. The cells were labeled with pY402 site-specific polyclonal antibody, followed by a secondary layer of TRITC-conjugated second antibody to rabbit IgG (red). Scale bar, 10 μm.

Techniques: Labeling, Phospho-proteomics, Expressing, Western Blot, Transfection, Positive Control, Control

( A ) Phosphorylation of Pyk2 Tyr-402 in PC12 cells transfected with Pyk2-related plasmids. PC12 cells were transfected with 4 μg of empty vectors (Mock), Pyk2-WT, Pyk2-Y402F, Pyk2-Y579F, Pyk2-Y580F, and Pyk2-Y881F for 48 h. The cells were treated with 1 μM ionomycin for 2 min, and immediately harvested for analysis of Pyk2 Tyr-402 phosphorylation by western blot. Actin was used as an internal control. The average Pyk2 phosphorylation in the Mock group was set as 100% for all experiments to standardize the results with the same plasmids from repeated experiments (n = 4/sample). ( B ) PC12 cells were transfected with 4 μg of empty vector (Mock), Pyk2-WT, Pyk2-Y402F, Pyk2-Y579F, Pyk2-Y580F, and Pyk2-Y881F for 48 h, and used for DA release assay with 1 μM ionomycin. The amount of DA released in the medium was expressed as the percentage of the total cellular content (n = 8/sample). The values are expressed as means ± S.E.M. from four representative experiments (*p<0.05, **p<0.01, ### p<0.001).

Journal: PLoS ONE

Article Title: Tyrosine 402 Phosphorylation of Pyk2 Is Involved in Ionomycin-Induced Neurotransmitter Release

doi: 10.1371/journal.pone.0094574

Figure Lengend Snippet: ( A ) Phosphorylation of Pyk2 Tyr-402 in PC12 cells transfected with Pyk2-related plasmids. PC12 cells were transfected with 4 μg of empty vectors (Mock), Pyk2-WT, Pyk2-Y402F, Pyk2-Y579F, Pyk2-Y580F, and Pyk2-Y881F for 48 h. The cells were treated with 1 μM ionomycin for 2 min, and immediately harvested for analysis of Pyk2 Tyr-402 phosphorylation by western blot. Actin was used as an internal control. The average Pyk2 phosphorylation in the Mock group was set as 100% for all experiments to standardize the results with the same plasmids from repeated experiments (n = 4/sample). ( B ) PC12 cells were transfected with 4 μg of empty vector (Mock), Pyk2-WT, Pyk2-Y402F, Pyk2-Y579F, Pyk2-Y580F, and Pyk2-Y881F for 48 h, and used for DA release assay with 1 μM ionomycin. The amount of DA released in the medium was expressed as the percentage of the total cellular content (n = 8/sample). The values are expressed as means ± S.E.M. from four representative experiments (*p<0.05, **p<0.01, ### p<0.001).

Techniques: Phospho-proteomics, Transfection, Western Blot, Control, Plasmid Preparation, Release Assay

( A ) PC12 cells were transfected with 4 μg of empty vectors (Mock), Pyk2-Y402F, Pyk2-Y579F, Pyk2-Y580F, and Pyk2-Y881F for 48 h, respectively. The cells were washed thrice with low-K + solution, and incubated for 20 min in low-K + solution with or without (Mock) 20 μM PP2 for 20 min. The cells were washed thrice, and sequentially incubated for 2 min in low-K + solution with 1 μM ionomycin. The amount of DA release in the medium was expressed as the percentage of the total cellular content (n = 8/sample). ( B ) Another parallel performed groups after stimulated by ionomycin for 2 min were immediately harvested, and lysed for western blot using phosphorylation site-specific antibodies against Tyr-402 (pY402), Tyr-579 (pY579), Tyr-580 (pY580), or Tyr-881 (pY881). Actin was used as an internal control (n = 4/sample). ( C ) Equal amount of cell lysates harvested at different incubation period were immunoprecipitated with anti-Src antibody, then immunoblotted with anti-phosphotyrosine antibody. The total amount of Src was used as an internal control. The values are expressed as means ± S.E.M. from four representative experiments (*p<0.05, # p<0.05, ## p<0.01, ### p<0.001).

Journal: PLoS ONE

Article Title: Tyrosine 402 Phosphorylation of Pyk2 Is Involved in Ionomycin-Induced Neurotransmitter Release

doi: 10.1371/journal.pone.0094574

Figure Lengend Snippet: ( A ) PC12 cells were transfected with 4 μg of empty vectors (Mock), Pyk2-Y402F, Pyk2-Y579F, Pyk2-Y580F, and Pyk2-Y881F for 48 h, respectively. The cells were washed thrice with low-K + solution, and incubated for 20 min in low-K + solution with or without (Mock) 20 μM PP2 for 20 min. The cells were washed thrice, and sequentially incubated for 2 min in low-K + solution with 1 μM ionomycin. The amount of DA release in the medium was expressed as the percentage of the total cellular content (n = 8/sample). ( B ) Another parallel performed groups after stimulated by ionomycin for 2 min were immediately harvested, and lysed for western blot using phosphorylation site-specific antibodies against Tyr-402 (pY402), Tyr-579 (pY579), Tyr-580 (pY580), or Tyr-881 (pY881). Actin was used as an internal control (n = 4/sample). ( C ) Equal amount of cell lysates harvested at different incubation period were immunoprecipitated with anti-Src antibody, then immunoblotted with anti-phosphotyrosine antibody. The total amount of Src was used as an internal control. The values are expressed as means ± S.E.M. from four representative experiments (*p<0.05, # p<0.05, ## p<0.01, ### p<0.001).

Techniques: Transfection, Incubation, Western Blot, Phospho-proteomics, Control, Immunoprecipitation

( A ) PC12 cells were transfected with 100 nM Src siRNA or control siRNA for 48 h, then harvested, and lysed for western blot using anti-Src antibody and anti-Pyk2 antibody. ( B ) PC12 cells were transfected with 100 nM Src siRNA or control siRNA for 48 h, and incubated with 1 μM ionomycin for 2 min. Cells were harvested at 0, 2, 4, and 6 min after ionomycin treatment, then immunoprecipitated with anti-Src antibody, then immunoblotted with anti-phosphotyrosine antibody. The total amount of Src was used as an internal control. ( C ) PC12 cells were transfected with 100 nM Src siRNA or control siRNA for 48 h, and incubated with 1 μM ionomycin for 2 min. Cells were harvested at 0, 2, 4, and 6 min after ionomycin treatment for western blot to test the expression of pY402 and total Pyk2. Pyk2 Y402 phosphorylation at each indicated time is expressed as a percentage of the maximal level of induced phosphorylation. ( D ) PC12 cells were transfected with 100 nM Src siRNA or control siRNA for 48 h, and incubated with 1 μM ionomycin for 2 min. The amount of DA released in the medium was expressed as the percentage of the total cellular content. The values are expressed as mean ± S.E.M. from four representative experiments (*p<0.05).

Journal: PLoS ONE

Article Title: Tyrosine 402 Phosphorylation of Pyk2 Is Involved in Ionomycin-Induced Neurotransmitter Release

doi: 10.1371/journal.pone.0094574

Figure Lengend Snippet: ( A ) PC12 cells were transfected with 100 nM Src siRNA or control siRNA for 48 h, then harvested, and lysed for western blot using anti-Src antibody and anti-Pyk2 antibody. ( B ) PC12 cells were transfected with 100 nM Src siRNA or control siRNA for 48 h, and incubated with 1 μM ionomycin for 2 min. Cells were harvested at 0, 2, 4, and 6 min after ionomycin treatment, then immunoprecipitated with anti-Src antibody, then immunoblotted with anti-phosphotyrosine antibody. The total amount of Src was used as an internal control. ( C ) PC12 cells were transfected with 100 nM Src siRNA or control siRNA for 48 h, and incubated with 1 μM ionomycin for 2 min. Cells were harvested at 0, 2, 4, and 6 min after ionomycin treatment for western blot to test the expression of pY402 and total Pyk2. Pyk2 Y402 phosphorylation at each indicated time is expressed as a percentage of the maximal level of induced phosphorylation. ( D ) PC12 cells were transfected with 100 nM Src siRNA or control siRNA for 48 h, and incubated with 1 μM ionomycin for 2 min. The amount of DA released in the medium was expressed as the percentage of the total cellular content. The values are expressed as mean ± S.E.M. from four representative experiments (*p<0.05).

Techniques: Transfection, Control, Western Blot, Incubation, Immunoprecipitation, Expressing, Phospho-proteomics

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anti human arc antibody (Proteintech)

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