Anti-SYT2 + SYT1 antibody [EPR23920-2]

9 product images

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  Western blot - Anti-SYT2 + SYT1 antibody [EPR23920-2] (ab259977)

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

  Unknown band at 37 kDa (PMID:32930942).

  Negative: human liver (PMID:17492637).

  Exposure time: 5.5 seconds

  All lanes: Western blot - Anti-SYT2 + SYT1 antibody [EPR23920-2] (ab259977) at 1/1000 dilution

  Lane 1: Human cerebellum tissue lysate at 20 µg

  Lane 2: Human liver tissue lysate at 20 µg

  Secondary

  All lanes: Western blot - VeriBlot for IP Detection Reagent (HRP) (VeriBlot for IP Detection Reagent (HRP) ab131366) at 1/1000 dilution

  Observed band size: 37 kDa, 60 kDa

  This data was developed using ab259977, the same antibody clone in a different buffer formulation.
  

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

  Unknown band at 37 kDa (PMID:32930942).

  Negative: human liver (PMID:17492637).

  Exposure time: 5.5 seconds

• 

  Western blot - Anti-SYT2 + SYT1 antibody [EPR23920-2] (ab259977)

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

  Unknown band at 37 kDa (PMID:32930942).

  Negative: mouse liver (PMID:17492637).

  Exposure time: Lane 1, 2, 4: 10 secondsLane 3: 3.3 seconds

  All lanes: Western blot - Anti-SYT2 + SYT1 antibody [EPR23920-2] (ab259977) at 1/1000 dilution

  Lane 1: PC-12 (rat adrenal gland pheochromocytoma) whole cell lysate at 20 µg

  Lane 2: C57 spinal cord tissue lysate at 20 µg

  Lane 3: Mouse brain tissue lysate at 20 µg

  Lane 4: Mouse liver tissue lysate at 20 µg

  Secondary

  All lanes: Western blot - Goat Anti-Rabbit IgG H&L (HRP) (Goat Anti-Rabbit IgG H&L (HRP) ab97051) at 1/20000 dilution

  Observed band size: 37 kDa, 48 kDa, 60 kDa

  This data was developed using ab259977, the same antibody clone in a different buffer formulation.
  

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

  Unknown band at 37 kDa (PMID:32930942).

  Negative: mouse liver (PMID:17492637).

  Exposure time: Lane 1, 2, 4: 10 secondsLane 3: 3.3 seconds

• 

  Western blot - Anti-SYT2 + SYT1 antibody [EPR23920-2] (ab259977)

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

  This antibody reacts with mouse SYT1 and SYT2 but not with SYT3, SYT4, SYT5, SYT6, SYT7, SYT9. These rec proteins were both made in house and extracted from E.coil expression systems.

  Exposure time: 15 seconds

  All lanes: Western blot - Anti-SYT2 + SYT1 antibody [EPR23920-2] (ab259977) at 1/1000 dilution

  Lane 1: His-tagged mouse recombinant protein SYT1 (aa137-421) (35 kDa) at 0.03 µg

  Lane 2: His-tagged mouse recombinant protein SYT2 (aa137-420) (34.8 kDa) at 0.03 µg

  Lane 3: His-tagged mouse recombinant protein SYT3 (aa 76-587) (56.9 kDa) at 0.03 µg

  Lane 4: His-tagged mouse recombinant protein SYT4 (aa 38-425) (45.6 kDa) at 0.03 µg

  Lane 5: His-tagged mouse recombinant protein SYT5 (aa 46-386) (40.4 kDa) at 0.03 µg

  Lane 6: His-tagged mouse recombinant protein SYT6 (aa 81-511) (51kDa) at 0.03 µg

  Lane 7: His-tagged mouse recombinant protein SYT7 (aa 38-403) (43.6 kDa) at 0.03 µg

  Lane 8: His-tagged mouse recombinant protein SYT9 (aa 74-491) (50.2 kDa) at 0.03 µg

  Secondary

  All lanes: Western blot - Goat Anti-Rabbit IgG H&L (HRP) (Goat Anti-Rabbit IgG H&L (HRP) ab97051) at 1/100000 dilution

  Observed band size: 34.8-56.9 kDa

  This data was developed using ab259977, the same antibody clone in a different buffer formulation.
  

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

  This antibody reacts with mouse SYT1 and SYT2 but not with SYT3, SYT4, SYT5, SYT6, SYT7, SYT9. These rec proteins were both made in house and extracted from E.coil expression systems.

  Exposure time: 15 seconds

• 

  Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-SYT2 + SYT1 antibody [EPR23920-2] (ab259977)

  Immunohistochemical analysis of paraffin-embedded Mouse cerebellum tissue labeling SYT1 + SYT2 with ab259977 at 1/50 dilution followed by a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (Rabbit specific IHC polymer detection kit HRP/DAB ab209101). Cytoplasmic staining on mouse cerebellum. The section was incubated with ab259977 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 Rabbit specific IHC polymer detection kit HRP/DAB (Rabbit specific IHC polymer detection kit HRP/DAB ab209101).

  Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution2) for 20 mins

• 

  Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-SYT2 + SYT1 antibody [EPR23920-2] (ab259977)

  Immunohistochemical analysis of paraffin-embedded Rat cerebellum tissue labeling SYT1 + SYT2 with ab259977 at 1/50 dilution followed by a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (Rabbit specific IHC polymer detection kit HRP/DAB ab209101). Cytoplasmic staining on rat cerebellum (PMID: 1856191). The section was incubated with ab259977 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 Rabbit specific IHC polymer detection kit HRP/DAB (Rabbit specific IHC polymer detection kit HRP/DAB ab209101).

  Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution2) for 20 mins

• 

  Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-SYT2 + SYT1 antibody [EPR23920-2] (ab259977)

  Immunohistochemical analysis of paraffin-embedded Mouse lung tissue labeling SYT1 + SYT2 with ab259977 at 1/50 dilution followed by a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (Rabbit specific IHC polymer detection kit HRP/DAB ab209101). Negative control: No staining on mouse lung (PMID: 1856191). The section was incubated with ab259977 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 Rabbit specific IHC polymer detection kit HRP/DAB (Rabbit specific IHC polymer detection kit HRP/DAB ab209101).

  Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution2) for 20 mins

• 

  Immunoprecipitation - Anti-SYT2 + SYT1 antibody [EPR23920-2] (ab259977)

  SYT1 + SYT2 was immunoprecipitated from 0.35 mg Human cerebellum tissue lysate 10 ug with ab259977 at 1/30 dilution (2ug in 0.35mg lysates). Western blot was performed on the immunoprecipitate using ab259977 at 1/1000 dilution. VeriBlot for IP Detection Reagent (HRP)(VeriBlot for IP Detection Reagent (HRP) ab131366) was used at 1/5000 dilution.
  

  Lane 1: Human cerebellum tissue lysate 10 ug

  Lane 2: ab259977 IP in Human cerebellum tissue lysate

  Lane 3: Rabbit monoclonal IgG (Rabbit IgG, monoclonal [EPR25A] - Isotype Control ab172730) instead of ab259977 in human cerebellum tissue lysate

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

  Exposure time: 3 minutes

  All lanes: Immunoprecipitation - Anti-SYT2 + SYT1 antibody [EPR23920-2] (ab259977)

  Observed band size: 60 kDa

• 

  Immunohistochemistry (Frozen sections) - Anti-SYT2 + SYT1 antibody [EPR23920-2] (ab259977)

  Immunohistochemical analysis of 4% PFA-fixed, 0.2% Triton X-100 permeabilized frozen Mouse hippocampus tissue labeling SYT1 + SYT2 with ab259977 at 1/50 dilution followed by Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) ab150077 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) at 1/1000 dilution (Green). Positive staining on mouse hippocampus is observed. The nuclear counterstain was DAPI (Blue).
  

  Secondary antibody control: Secondary antibody is Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) ab150077 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488)at 1/1000 dilution.

  Heat mediated antigen retrieval using sodium citrate buffer (10mM citrate pH 6.0 + 0.05% Tween-20).

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  Immunocytochemistry/ Immunofluorescence - Anti-SYT2 + SYT1 antibody [EPR23920-2] (ab259977)

  Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% TritonX-100 permeabilized mouse primary neuron cells labelling SYT1 + SYT2 with ab259977 at 1/50 dilution, followed by Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) ab150077 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) antibody at 1/1000 dilution (Green). Confocal image showing positive staining in synapse of mouse primary neurons. Confocal scanning Z step was set as 0.3 μm followed by image processing with maximum Z projection is observed. Anti-MAP2 antibody [HM-2] ab11267 Anti-MAP2 mouse monoclonal antibody was used to counterstain tubulin at 1/500 followed by Goat Anti-Mouse IgG H&L (Alexa Fluor® 594) preadsorbed ab150120 Goat Anti-Mouse IgG H&L (Alexa Fluor® 594) at 1/1000 dilution (Red). The Nuclear counterstain was DAPI (Blue).
  

  Secondary antibody only control: Secondary antibody is Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) ab150077 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) at 1/1000 dilution.