Anti-Synaptogyrin 3 抗体 [EPR25082-15] (ab302613)

Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% TritonX-100 permeabilized mouse primary neural/glia cells labeling Synaptogyrin 3 with ab302613 at 1/50 (10.08 µg/ml) dilution, followed by ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor^® 488) preadsorbed antibody at 1/1000 2µg/ml dilution (Green). Confocal image showing synaptic staining in mouse primary neural/glia cells. Confocal scanning Z step was set as 0.3 µm followed by image processing with maximum Z projection. Negative control: C2C12. ab11267 Anti-MAP2 mouse monoclonal antibody was used to counterstain tubulin at 1/500 (4µg/ml) dilution (Red). The Nuclear counterstain was DAPI (Blue).

Negative control 1: ab302613 at 1/50 dilution (10.08 μg/ml) with ab150120 (Counterstain Secondary) as secondary antibody at 1/1000 dilution (2μg/ml)

Negative control 2: ab11267 at 1/500 dilution (4 μg/ml) with ab150081 (Target Secondary) as secondary antibody at 1/1000 dilution (2μg/ml)

Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% TritonX-100 permeabilized rat primary neural/glia cells labeling Synaptogyrin 3 with ab302613 at 1/50 (10.08 µg/ml) dilution, followed by ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor^® 488) preadsorbed antibody at 1/1000 (2µg/ml) dilution (Green). Confocal image showing synaptic staining in rat primary neural/glia cells. Confocal scanning Z step was set as 0.3 µm followed by image processing with maximum Z projection. ab11267 Anti-MAP2 mouse monoclonal antibody was used to counterstain tubulin at 1/500 4µg/ml dilution (Red). The Nuclear counterstain was DAPI (Blue).

Negative control 1: ab302613 at 1/50 dilution (10.08 μg/ml) with ab150120 (Counterstain Secondary) as secondary antibody at 1/1000 dilution (2μg/ml)

Negative control 2: ab11267 at 1/500 dilution (4 μg/ml) with ab150081 (Target Secondary) as secondary antibody at 1/1000 dilution (2μg/ml)

Immunohistochemical analysis of paraffin-embedded mouse hippocampus tissue labeling Synaptogyrin 3 with ab302613 at 1/5000 (0.101 µg/ml) followed by a ready to use Leica DS9800 (Bond™ Polymer Refine Detection kit). Positive staining on mouse hippocampus (PMID:33472038). The section was incubated with ab302613 for 30 mins at room temperature. The immunostaining was performed on a Leica Biosystems BOND^® RX instrument counterstained with Hematoxylin.

Secondary antibody only control: Secondary antibody is a ready to use Leica DS9800 (Bond™ Polymer Refine Detection kit).

Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution 2) for 20 mins

Immunohistochemical analysis of paraffin-embedded rat cerebrum tissue labeling Synaptogyrin 3 with ab302613 at 1/5000 (0.101 µg/ml) followed by a ready to use Leica DS9800 (Bond™ Polymer Refine Detection kit). Positive staining on rat cerebrum. The section was incubated with ab302613 for 30 mins at room temperature. The immunostaining was performed on a Leica Biosystems BOND^® RX instrument counterstained with Hematoxylin.

Secondary antibody only control: Secondary antibody is a ready to use Leica DS9800 (Bond™ Polymer Refine Detection kit).

Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution 2) for 20 mins

Immunohistochemical analysis of paraffin-embedded mouse pancreas tissue labeling Synaptogyrin 3 with ab302613 at 1/5000 (0.101 µg/ml) followed by a ready to use Leica DS9800 (Bond™ Polymer Refine Detection kit). Negative control: No staining on mouse pancreas. The section was incubated with ab302613 for 30 mins at room temperature. The immunostaining was performed on a Leica Biosystems BOND^® RX instrument counterstained with Hematoxylin.

Secondary antibody only control: Secondary antibody is a ready to use Leica DS9800 (Bond™ Polymer Refine Detection kit).

Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution 2) for 20 mins

Immunohistochemical analysis of 4% PFA-fixed, 0.2% Triton X-100 permeabilized frozen mouse cerebrum (fresh) tissue labeling Synaptogyrin 3 with ab302613 at 1/500 (1.008 µg/ml) dilution followed by ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor^® 488) preadsorbed at 1/1000 (2 µg/ml) dilution (Green). Positive staining on mouse cerebrum is observed. The nuclear counterstain was DAPI (Blue).

Secondary antibody control: Secondary antibody is ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor^® 488) preadsorbed at 1/1000 (2 µg/ml) dilution.

Immunohistochemical analysis of 4% PFA-fixed, 0.2% Triton X-100 permeabilized frozen rat cerebrum (fresh) tissue labeling Synaptogyrin 3 with ab302613 at 1/500 (1.008 µg/ml) dilution followed by ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor^® 488) preadsorbed at 1/1000 (2 µg/ml) dilution (Green). Positive staining on rat cerebrum is observed. The nuclear counterstain was DAPI (Blue).

Secondary antibody control: Secondary antibody is ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor^® 488) preadsorbed at 1/1000 (2 µg/ml) dilution.

All lanes : Anti-Synaptogyrin 3 antibody [EPR25082-15] (ab302613) at 1/1000 dilution

Lane 1 : Human eyeball tissue lysate
Lane 2 : Human hypothalamus tissue lysate
Lane 3 : Human pancreas tissue lysate

Lysates/proteins at 50 µg per lane.

Secondary
All lanes : Goat Anti-Rabbit IgG (HRP) with minimal cross-reactivity with human IgG at 1/1000 dilution

Predicted band size: 24 kDa
Observed band size: 26 kDa why is the actual band size different from the predicted?

Blocking / Diluting buffer and concentration: 5% NFDM/TBST

Exposure time:

Lane 1 and 3: 48 seconds
Lane 2: 5.5 seconds

Negative control: pancreas (Database :HPA, the human protein atlas)

All lanes : Anti-Synaptogyrin 3 antibody [EPR25082-15] (ab302613) at 1/1000 dilution

Lane 1 : Mouse brain tissue lysate
Lane 2 : Mouse hypothalamus tissue lysate
Lane 3 : Mouse pancreas tissue lysate

Lysates/proteins at 50 µg per lane.

Secondary
All lanes : Goat Anti-Rabbit IgG (HRP) with minimal cross-reactivity with human IgG at 1/1000 dilution

Predicted band size: 24 kDa
Observed band size: 26 kDa why is the actual band size different from the predicted?


Exposure time: 1 second

Blocking / Diluting buffer and concentration: 5% NFDM/TBST

Negative control: pancreas (Database :HPA, the human protein atlas)

All lanes : Anti-Synaptogyrin 3 antibody [EPR25082-15] (ab302613) at 1/1000 dilution

Lane 1 : Neuro-2a (mouse neuroblastoma neuroblast), whole cell lysate
Lane 2 : C2C12 (mouse myoblasts myoblast), whole cell lysate
Lane 3 : SH-SY5Y (human neuroblastoma epithelial cell), whole cell lysate
Lane 4 : 293T (human embryonic kidney epithelial cell), whole cell lysate

Lysates/proteins at 50 µg per lane.

Secondary
All lanes : Goat Anti-Rabbit IgG (HRP) with minimal cross-reactivity with human IgG at 1/1000 dilution

Predicted band size: 24 kDa
Observed band size: 26 kDa why is the actual band size different from the predicted?

Blocking / Diluting buffer and concentration: 5% NFDM/TBST

Exposure time:

Lane 1: 48 seconds
Lane 2, 3 and 4: 136 seconds

Negative controls: 293T (PMID: 14755516), C2C12

All lanes : Anti-Synaptogyrin 3 antibody [EPR25082-15] (ab302613) at 1/1000 dilution

Lane 1 : Rat hypothalamus tissue lysate
Lane 2 : Rat pancreas tissue lysate

Lysates/proteins at 50 µg per lane.

Secondary
All lanes : Goat Anti-Rabbit IgG (HRP) with minimal cross-reactivity with human IgG at 1/1000 dilution

Predicted band size: 24 kDa
Observed band size: 26 kDa why is the actual band size different from the predicted?


Exposure time: 1 second

Blocking / Diluting buffer and concentration: 5% NFDM/TBST

Negative control: pancreas (Database :HPA, the human protein atlas)