Anti-Synaptogyrin 1 抗体 [EPR25081-18]

• IHC-P

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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Synaptogyrin 1 antibody [EPR25081-18] (AB308617)

Immunohistochemical analysis of paraffin-embedded Human hypothalamus tissue labelling Synaptogyrin 1 with ab308617 at 1/5000 (0.1 ug/ml) followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection). Cytoplasmic staining on human hypothalamus. The section was incubated with ab308617 for 10 mins at room temperature. The immunostaining was performed on a Leica Biosystems BOND® RX instrument. Incubate slides with 3% Hydrogen Peroxide for 10 mins at room temperature after secondary antibody incubation to reduce the background. Counterstained with Hematoxylin. Secondary antibody only control : Secondary antibody is a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection). Heat mediated antigen retrieval was performed with Tris-EDTA buffer (pH 9.0, Epitope Retrieval Solution2) for 20 mins

• IHC-P

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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Synaptogyrin 1 antibody [EPR25081-18] (AB308617)

Immunohistochemical analysis of paraffin-embedded Human lung tissue labelling Synaptogyrin 1 with ab308617 at 1/5000 (0.1 ug/ml) followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection). Negative control : No staining on human lung. The section was incubated with ab308617 for 10 mins at room temperature. The immunostaining was performed on a Leica Biosystems BOND® RX instrument. Incubate slides with 3% Hydrogen Peroxide for 10 mins at room temperature after secondary antibody incubation to reduce the background. Counterstained with Hematoxylin. Secondary antibody only control : Secondary antibody is a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection). Heat mediated antigen retrieval was performed with Tris-EDTA buffer (pH 9.0, Epitope Retrieval Solution2) for 20 mins

• IHC-P

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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Synaptogyrin 1 antibody [EPR25081-18] (AB308617)

Immunohistochemical analysis of paraffin-embedded Human cerebrum tissue labelling Synaptogyrin 1 with ab308617 at 1/5000 (0.1 ug/ml) followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection). Cytoplasmic staining on human cerebrum (PMID : 10595519). The section was incubated with ab308617 for 10 mins at room temperature. The immunostaining was performed on a Leica Biosystems BOND® RX instrument. Incubate slides with 3% Hydrogen Peroxide for 10 mins at room temperature after secondary antibody incubation to reduce the background. Counterstained with Hematoxylin. Secondary antibody only control : Secondary antibody is a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection). Heat mediated antigen retrieval was performed with Tris-EDTA buffer (pH 9.0, Epitope Retrieval Solution2) for 20 mins

• IP

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Immunoprecipitation - Anti-Synaptogyrin 1 antibody [EPR25081-18] (AB308617)

Synaptogyrin 1 was immunoprecipitated from 0.35 mg Human hypothalamus tissue lysate with ab308617 at 1/30 dilution (2ug in 0.35mg lysates). Western blot was performed on the immunoprecipitate using ab308617 at 1/1000 dilution. VeriBlot for IP secondary antibody (HRP) (ab131366) was used at 1/5000 dilution.

Lane 1 : Human hypothalamus tissue lysate
Lane 2 : ab308617 IP in Human hypothalamus tissue lysate
Lane 3 : Rabbit monoclonal IgG (ab172730) instead of ab308617 in human hypothalamus tissue lysate

Blocking and dilution buffer and concentration : 5% NFDM/TBST.

Exposure time : 1 second

All lanes:

Immunoprecipitation - Anti-Synaptogyrin 1 antibody [EPR25081-18] (ab308617) at 1/1000 dilution

All lanes:

Human hypothalamus tissue lysate at 10 µg

Secondary

All lanes:

Immunoprecipitation - VeriBlot for IP Detection Reagent (HRP) (<a href='/products/reagents/veriblot-for-ip-detection-reagent-hrp-ab131366'>ab131366</a>) at 1/5000 dilution

Observed band size: 26 kDa

false

Exposure time: 1s

• Flow Cyt (Intra)

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Flow Cytometry (Intracellular) - Anti-Synaptogyrin 1 antibody [EPR25081-18] (AB308617)

Flow cytometric analysis of 4% paraformaldehyde fixed 90% methanol permeabilized Rat primary neuron cells labelling Synaptogyrin 1 with ab308617 at 1/500 dilution (0.1 ug)/Right (Red) compared with a Rabbit monoclonal IgG (ab172730) / Left isotype control and an unlabelled control (cells without incubation with primary antibody and secondary antibody) (Blue). A Goat Anti-Rabbit IgG (Alexa Fluor® 488, ab150081) at 1/5000 dilution was used as the secondary antibody.

• Flow Cyt (Intra)

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Flow Cytometry (Intracellular) - Anti-Synaptogyrin 1 antibody [EPR25081-18] (AB308617)

Flow cytometric analysis of 4% paraformaldehyde fixed 90% methanol permeabilized Mouse primary neuron cells labelling Synaptogyrin 1 with ab308617 at 1/500 dilution (0.1 ug)/Right (Red) compared with a Rabbit monoclonal IgG (ab172730) / Left isotype control and an unlabelled control (cells without incubation with primary antibody and secondary antibody) (Blue). A Goat Anti-Rabbit IgG (Alexa Fluor® 488, ab150081) at 1/5000 dilution was used as the secondary antibody.

• ICC/IF

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Immunocytochemistry/ Immunofluorescence - Anti-Synaptogyrin 1 antibody [EPR25081-18] (AB308617)

Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% TritonX-100 permeabilized rat primary neural/glia cell cells labelling Synaptogyrin 1 with ab308617 at 1/100 (4.985 ug/ml) dilution, followed by ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed antibody at 1/1000 2 ug/ml dilution (Green). Confocal image showing positive staining in rat primary neural/glia cell. Confocal scanning Z step was set as 0.3 µm followed by image processing with maximum Z projection. Image was taken with a confocal microscope (Leica-Microsystems, TCS SP8). ab11267 Anti-MAP2 mouse monoclonal antibody was used to counterstain tubulin at 1/500 4ug/ml dilution (Red). The Nuclear counterstain was DAPI (Blue). Secondary antibody only control : Secondary antibody is ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed at 1/1000 2 ug/ml dilution.

• ICC/IF

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Immunocytochemistry/ Immunofluorescence - Anti-Synaptogyrin 1 antibody [EPR25081-18] (AB308617)

Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% TritonX-100 permeabilized mouse primary neural/glia cell cells labelling Synaptogyrin 1 with ab308617 at 1/100 (4.985 ug/ml) dilution, followed by ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed antibody at 1/1000 2 ug/ml dilution (Green). Confocal image showing positive staining in mouse primary neural/glia cell. Confocal scanning Z step was set as 0.3 µm followed by image processing with maximum Z projection. Image was taken with a confocal microscope (Leica-Microsystems, TCS SP8). ab11267 Anti-MAP2 mouse monoclonal antibody was used to counterstain tubulin at 1/500 4ug/ml dilution (Red). The Nuclear counterstain was DAPI (Blue). Secondary antibody only control : Secondary antibody is ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed at 1/1000 2 ug/ml dilution.

• IHC-P

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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Synaptogyrin 1 antibody [EPR25081-18] (AB308617)

Immunohistochemical analysis of paraffin-embedded Mouse cerebrum tissue labelling Synaptogyrin 1 with ab308617 at 1/5000 (0.1 ug/ml) followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection). Cytoplasmic staining on mouse cerebrum.The section was incubated with ab308617 for 10 mins at room temperature. The immunostaining was performed on a Leica Biosystems BOND® RX instrument. Incubate slides with 3% Hydrogen Peroxide for 10 mins at room temperature after secondary antibody incubation to reduce the background. Counterstained with Hematoxylin. Secondary antibody only control : Secondary antibody is a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection). Heat mediated antigen retrieval was performed with Tris-EDTA buffer (pH 9.0, Epitope Retrieval Solution2) for 20 mins

• IHC-P

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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Synaptogyrin 1 antibody [EPR25081-18] (AB308617)

Immunohistochemical analysis of paraffin-embedded Rat cerebrum tissue labelling Synaptogyrin 1 with ab308617 at 1/5000 (0.1 ug/ml) followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection). Cytoplasmic staining on rat cerebrum (PMID : 9446595). The section was incubated with ab308617 for 10 mins at room temperature. The immunostaining was performed on a Leica Biosystems BOND® RX instrument. Incubate slides with 3% Hydrogen Peroxide for 10 mins at room temperature after secondary antibody incubation to reduce the background. Counterstained with Hematoxylin. Secondary antibody only control : Secondary antibody is a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection). Heat mediated antigen retrieval was performed with Tris-EDTA buffer (pH 9.0, Epitope Retrieval Solution2) for 20 mins

• WB

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Western blot - Anti-Synaptogyrin 1 antibody [EPR25081-18] (AB308617)

Blocking and diluting buffer and concentration : 5% NFDM/TBST. Negative control : lung, liver (PMID : 9446595). In Western blot, anti- Vinculin antibody (ab129002) loading control staining at 1/10, 000 dilution. Exposure time : 3 seconds

All lanes:

Western blot - Anti-Synaptogyrin 1 antibody [EPR25081-18] (ab308617) at 1/1000 dilution

Lane 1:

Human hypothalamus tissue lysate at 20 µg

Lane 2:

Human lung tissue lysate at 20 µg

Lane 3:

Human liver tissue lysate at 20 µg

Lane 4:

Mouse brain tissue lysate at 20 µg

Lane 5:

Mouse hypothalamus tissue lysate at 20 µg

Lane 6:

Mouse lung tissue lysate at 20 µg

Lane 7:

Rat brain tissue lysate at 20 µg

Lane 8:

Rat lung tissue lysate at 20 µg

Lane 9:

Rat hypothalamus tissue lysate at 20 µg

Secondary

Lanes 1 - 9:

Western blot - Goat Anti-Rabbit IgG H&L (HRP) (<a href='/products/secondary-antibodies/goat-rabbit-igg-h-l-hrp-ab97051'>ab97051</a>) at 1/2000 dilution

Lanes 1 - 9:

Goat Anti-Rabbit IgG (HRP) with minimal cross-reactivity with human IgG at 1/20000 dilution

Predicted band size: 25 kDa

Observed band size: 26 kDa

false

Exposure time: 3s

• WB

Supplier Data

Western blot - Anti-Synaptogyrin 1 antibody [EPR25081-18] (AB308617)

Blocking and diluting buffer and concentration : 5% NFDM/TBST. This antibody does not cross-react with human Synaptogyrin 2. In Western blot, anti-His antibody (ab213204) staining at 1/5000 dilution, anti-GAPDH antibody (ab181602) loading control staining at 1/200, 000 dilution. Exposure time : 10 seconds

All lanes:

Western blot - Anti-Synaptogyrin 1 antibody [EPR25081-18] (ab308617) at 1/1000 dilution

Lane 1:

HEK-293 (human embryonic kidney) transfected with an empty vector (vector control), containing a myc-His-tag®, whole cell lysate at 20 µg

Lane 2:

HEK-293 transfected with human Synaptogyrin 2 (WT) expression vector containing a myc-His-tag®, whole cell lysate at 20 µg

Lane 3:

His-tagged human Synaptogyrin 1 fragment at 10 ng

Secondary

All lanes:

Western blot - Goat Anti-Rabbit IgG H&L (HRP) (<a href='/products/secondary-antibodies/goat-rabbit-igg-h-l-hrp-ab97051'>ab97051</a>) at 1/100000 dilution

Predicted band size: 25 kDa

Observed band size: 30 kDa

false

Exposure time: 10s