Anti-Rab3A antibody [EPR26022-9]

20 product images

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  Western blot - Anti-Rab3A antibody [EPR26022-9] (ab302518)

  ab302518 was shown to react with RAB3A in wild-type SKNFI cells in Western blot with loss of signal observed in a RAB3A knockout cell line. Wild-type SKNFI and RAB3A knockout cell lysates were subjected to SDS-PAGE. Membranes were blocked in 5% milk in TBST for 1 hr before incubation with ab302518 overnight at 4 °C at a 1/1000 dilution. Blots were incubated with secondary antibodies at 0.2 µg/mL before imaging.
  
  These data were provided by YCharOS Inc., an open science company with the mission of characterizing commercially available antibody reagents for all human proteins. Abcam and YCharOS are working together to help address the reproducibility crisis by enabling the life science community to better evaluate commercially available antibodies.

  All lanes: Western blot - Anti-Rab3A antibody [EPR26022-9] (ab302518) at 1/1000 dilution

  Lane 1: Wild-type SKNFI lysate at 40 µg

  Lane 2: RAB3A knock-out SKNFI lysate at 40 µg

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  Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Rab3A antibody [EPR26022-9] (ab302518)

  Immunohistochemical analysis of paraffin-embedded (A) Cerebellum tissue from wild-type C57BL/6JGpt mice (B) Cerebellum tissue from Rab3A knockout mice labeling Rab3A with ab302518 at 1/5000 dilution (0.11 µg/ml) followed by a ready to use LeicaDS9800 (BOND™ Polymer Refine Detection). Sections were fixed and
  counterstained with Hematoxylin. Antigen retrieval was Heat mediated with Citrate buffer (pH 6.0, Epitope Retrieval Solution 1) for 20 mins.
  
  Positive staining on (A) Cerebellum tissue from wild-type C57BL/6JGpt mice and no staining on (B) Cerebellum tissue from Rab3A knockout mice.
  The section was incubated with ab302518 for 30 mins at room temperature.
  The immunostaining was performed on a Leica Biosystems BOND® RX instrument.
  The tissue samples were kindly provided by GemPharmatech. C57BL/6JGpt wildtype mice and Rab3A-KO homozygous mice (Strain ID: T016905).

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  Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Rab3A antibody [EPR26022-9] (ab302518)

  Immunohistochemical analysis of paraffin-embedded (A) Brain cortex tissue from wild-type C57BL/6JGpt mice (B) Brain cortex tissue from Rab3A knockout mice labeling Rab3A with ab302518 at 1/5000 dilution (0.11 µg/ml) followed by a ready to use LeicaDS9800 (BOND™ Polymer Refine Detection). Sections were fixed and counterstained with Hematoxylin. Antigen retrieval was Heat mediated with Citrate buffer (pH 6.0, Epitope Retrieval Solution 1) for 20 mins.
  
  Positive staining on (A) Brain cortex tissue from wild-type C57BL/6JGpt mice and no staining on (B) Brain cortex tissue from Rab3A knockout mice.
  The section was incubated with ab302518 for 30 mins at room temperature.
  The immunostaining was performed on a Leica Biosystems BOND® RX instrument.
  The tissue samples were kindly provided by GemPharmatech. C57BL/6JGpt wildtype mice and Rab3A-KO homozygous mice (Strain ID: T016905).

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  Western blot - Anti-Rab3A antibody [EPR26022-9] (ab302518)

  Loading control: Anti-GAPDH antibody [EPR16891] - Loading Control (Anti-GAPDH antibody [EPR16891] - Loading Control ab181602) (1:200000) (36KDa)
  
  Blocking/Diluting buffer and concentration: 5% NFDM/TBST
  
  The tissue samples were kindly provided by GemPharmatech. C57BL/6JGpt wildtype mice and Rab3A-KO homozygous mice (Strain ID: T016905).

  Lanes 1 - 12: Western blot - Anti-Rab3A antibody [EPR26022-9] (ab302518) at 1/1000 dilution

  Lanes 1 - 12: Western blot - Anti-Rab3A antibody [EPR26022-9] - BSA and Azide free (Anti-Rab3A antibody [EPR26022-9] - BSA and Azide free ab302519)

  Lane 1: Wild-type mouse cerebellum tissue lysate (male)

  Lane 2: Rab3A knockout cerebellum tissue lysate (male)

  Lane 3: Wild-type mouse brain cortex tissue lysate (male)

  Lane 4: Rab3A knockout brain cortex tissue lysate (male)

  Lane 5: Wild-type mouse striatum tissue lysate (male)

  Lane 6: Rab3A knockout striatum tissue lysate (male)

  Lane 7: Wild-type mouse cerebellum tissue lysate (female)

  Lane 8: Rab3A knockout cerebellum tissue lysate (female)

  Lane 9: Wild-type mouse brain cortex tissue lysate (female)

  Lane 10: Rab3A knockout brain cortex tissue lysate (female)

  Lane 11: Wild-type mouse striatum tissue lysate (female)

  Lane 12: Rab3A knockout striatum tissue lysate (female)

  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: 25 kDa

  Exposure time: 1s

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  Western blot - Anti-Rab3A antibody [EPR26022-9] (ab302518)

  Western blot: Left: Anti-RAB3A antibody (Anti-Rab3A antibody ab3335) staining at 2 ug/ml. Right: Recombinant Anti-Rab3A antibody [EPR26022-9] (ab302518) staining at 1/1000 dilution. Anti-Rab3A antibodies shown in green; Mouse anti-Alpha Tubulin [DM1A] (Anti-alpha Tubulin antibody [DM1A] - Loading Control ab7291) loading control staining at 1/20000 dilution, shown in magenta. In Western blot, Anti-Rab3A antibody ab3335 and ab302518 were shown to bind specifically to RAB3A. A band was observed at 27 kDa in wild-type SK-N-FI cell lysates with no signal observed at this size in RAB3A knockout cell line. To generate this image, wild-type and RAB3A knockout SK-N-FI cell lysates were analysed. First, samples were run on an SDS-PAGE gel then transferred onto a nitrocellulose membrane. Membranes were blocked in 3% milk in TBS-0.1% Tween® 20 (TBS-T) before incubation with primary antibodies overnight at 4°C. Blots were washed four times in TBS-T, incubated with secondary antibodies for 1 h at room temperature, washed again four times then imaged. Secondary antibodies used were Goat anti-Rabbit IgG H&L 800CW and Goat anti-Mouse IgG H&L 680RD at 1/20000 dilution.

  Lanes 1 - 4: Western blot - Anti-Rab3A antibody (Anti-Rab3A antibody ab3335) at 2 µg/mL

  Lanes 1 - 4: Western blot - Anti-Rab3A antibody [EPR26022-9] (ab302518) at 1/1000 dilution

  Lane 1: Wild-type SK-N-FI cell lysate at 20 µg

  Lane 2: RAB3A knockout SK-N-FI cell lysate at 20 µg

  Lane 3: Human brain cell lysate at 20 µg

  Lane 4: HEK-293 cell lysate at 20 µg

  Developed using the ECL technique.

  Performed under reducing conditions.

  Observed band size: 27 kDa

• 

  Western blot - Anti-Rab3A antibody [EPR26022-9] (ab302518)

  Anti-RAB3A antibody [EPR26022-9] (ab302518) staining at 1/1000 dilution, shown in green; Mouse anti-Alpha Tubulin [DM1A] (Anti-alpha Tubulin antibody [DM1A] - Loading Control ab7291) loading control staining at 1/20000 dilution, shown in red. In Western blot, ab302518 was shown to bind specifically to RAB3A. A band was observed at 27 kDa in wild-type SKNFI cell lysates with no signal observed at this size in RAB3A knockout cell line. To generate this image, wild-type and RAB3A knockout SKNFI cell lysates were analysed. First, samples were run on an SDS-PAGE gel then transferred onto a nitrocellulose membrane. Membranes were blocked in 3% milk in TBS-0.1% Tween® 20 (TBS-T) before incubation with primary antibodies overnight at 4 °C. Blots were washed four times in TBS-T, incubated with secondary antibodies for 1 h at room temperature, washed again four times then imaged. Secondary antibodies used were Goat anti-Rabbit IgG H&L 800CW and Goat anti-Mouse IgG H&L 680RD at 1/20000 dilution.

  Lanes 1 - 4: Western blot - Anti-Rab3A antibody [EPR26022-9] (ab302518) at 1/1000 dilution

  Lanes 1 - 4: Western blot - Anti-Rab3A antibody [EPR26022-9] - BSA and Azide free (Anti-Rab3A antibody [EPR26022-9] - BSA and Azide free ab302519)

  Lane 1: Wild-type SKNFI cell lysate at 20 µg

  Lane 2: RAB3A knockout SKNFI cell lysate at 20 µg

  Lane 3: Human brain cell lysate at 20 µg

  Lane 4: HEK-293 cell lysate at 20 µg

  Performed under reducing conditions.

  Observed band size: 27 kDa

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  Dot Blot - Anti-Rab3A antibody [EPR26022-9] (ab302518)

  Dot blot analysis of Rab3A using ab302518 at 1:1000 (0.55 ug/ml) followed by a Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated (Goat Anti-Rabbit IgG H&L (HRP) ab97051) at 1:100,000 dilution. Exposure time: 3 minutes Blocking and diluting buffer and concentration: 5% NFDM/TBST

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  Immunohistochemistry (Frozen sections) - Anti-Rab3A antibody [EPR26022-9] (ab302518)

  Immunohistochemical analysis of 4% PFA-fixed, 0.2% Triton X-100 permeabilized frozen Rat cerebellum (fresh) tissue labeling Rab3A with ab302518 at 1/500 (1.1 ug/ml) dilution followed by Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed at 1/1000 2 ug/mL dilution (Green). Positive staining on rat cerebellum is observed. The nuclear counterstain was DAPI (Blue). Secondary antibody control: Secondary antibody is Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbedat 1/1000 2 ug/mL dilution.<\p> .

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  Immunohistochemistry (Frozen sections) - Anti-Rab3A antibody [EPR26022-9] (ab302518)

  Immunohistochemical analysis of 4% PFA-fixed, 0.2% Triton X-100 permeabilized frozen Mouse cerebellum (fresh) tissue labeling Rab3A with ab302518 at 1/500 (1.1 ug/ml) dilution followed by Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed at 1/1000 2 ug/mL dilution (Green). Positive staining on mouse cerebellum is observed. The nuclear counterstain was DAPI (Blue). Secondary antibody control: Secondary antibody is Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbedat 1/1000 2 ug/mL dilution.<\p> .

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  Flow Cytometry (Intracellular) - Anti-Rab3A antibody [EPR26022-9] (ab302518)

  Flow cytometric analysis of 4% paraformaldehyde fixed 90% methanol permeabilized PC-12 (rat adrenal gland pheochromocytoma) cells labelling Rab3A with ab302518 at 1/500 dilution (0.1ug) (Red) (Red) compared with a Rabbit monoclonal IgG (Rabbit IgG, monoclonal [EPR25A] - Isotype Control ab172730) (Black) isotype control and an unlabelled control (cells without incubation with primary antibody and secondary antibody) (Blue). A Goat Anti-Rabbit IgG (Alexa Fluor® 488, Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed ab150081) at 1/2000 dilution was used as the secondary antibody.

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  Flow Cytometry (Intracellular) - Anti-Rab3A antibody [EPR26022-9] (ab302518)

  Flow cytometric analysis of 4% paraformaldehyde fixed 90% methanol permeabilized Neuro-2a (mouse neuroblastoma neuroblast) cells labelling Rab3A with ab302518 at 1/500 dilution (0.1ug) (Red) (Red) compared with a Rabbit monoclonal IgG (Rabbit IgG, monoclonal [EPR25A] - Isotype Control ab172730) (Black) isotype control and an unlabelled control (cells without incubation with primary antibody and secondary antibody) (Blue). A Goat Anti-Rabbit IgG (Alexa Fluor® 488, Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed ab150081) at 1/2000 dilution was used as the secondary antibody.

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  Flow Cytometry (Intracellular) - Anti-Rab3A antibody [EPR26022-9] (ab302518)

  Flow cytometric analysis of 4% paraformaldehyde fixed 90% methanol permeabilized SH-SY5Y (human neuroblastoma epithelial cell) cells labelling Rab3A with ab302518 at 1/500 dilution (0.1ug) (Red) (Red) compared with a Rabbit monoclonal IgG (Rabbit IgG, monoclonal [EPR25A] - Isotype Control ab172730) (Black) isotype control and an unlabelled control (cells without incubation with primary antibody and secondary antibody) (Blue). A Goat Anti-Rabbit IgG (Alexa Fluor® 488, Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed ab150081) at 1/2000 dilution was used as the secondary antibody.

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  Immunocytochemistry - Anti-Rab3A antibody [EPR26022-9] (ab302518)

  Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% TritonX-100 permeabilized PC-12 (rat adrenal gland pheochromocytoma small irregularly shaped cells) cells labelling Rab3A with ab302518 at 1/50 (11.0 ug/ml) dilution, followed by Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed antibody at 1/1000 2ug/ml dilution (Green). Confocal image showing cytoplasmic staining in PC-12 cell line is observed. Alexa Fluor® 594 Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker ab195889 Anti-alpha Tubulin mouse monoclonal antibody - Microtubule Marker (Alexa Fluor® 594) was used to counterstain tubulin at 1/200 2.5ug/ml dilution (Red). The Nuclear counterstain was DAPI (Blue). Secondary antibody only control: Secondary antibody is Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed at 1/1000 2ug/ml dilution.

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  Immunocytochemistry - Anti-Rab3A antibody [EPR26022-9] (ab302518)

  Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% TritonX-100 permeabilized Neuro-2a (mouse neuroblastoma neuroblast) cells labelling Rab3A with ab302518 at 1/50 (11.0 ug/ml) dilution, followed by Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed antibody at 1/1000 2ug/ml dilution (Green). Confocal image showing cytoplasmic staining in Neuro-2a cell line is observed. Alexa Fluor® 594 Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker ab195889 Anti-alpha Tubulin mouse monoclonal antibody - Microtubule Marker (Alexa Fluor® 594) was used to counterstain tubulin at 1/200 2.5ug/ml dilution (Red). The Nuclear counterstain was DAPI (Blue). Secondary antibody only control: Secondary antibody is Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed at 1/1000 2ug/ml dilution.

• 

  Immunocytochemistry - Anti-Rab3A antibody [EPR26022-9] (ab302518)

  Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% TritonX-100 permeabilized SH-SY5Y (human neuroblastoma epithelial cell) cells labelling Rab3A with ab302518 at 1/50 (11.0 ug/ml) dilution, followed by Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed antibody at 1/1000 2ug/ml dilution (Green). Confocal image showing cytoplasmic staining in SH-SY5Y cell line is observed. Alexa Fluor® 594 Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker ab195889 Anti-alpha Tubulin mouse monoclonal antibody - Microtubule Marker (Alexa Fluor® 594) was used to counterstain tubulin at 1/200 2.5ug/ml dilution (Red). The Nuclear counterstain was DAPI (Blue). Secondary antibody only control: Secondary antibody is Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed at 1/1000 2ug/ml dilution.

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  Immunohistochemistry - Anti-Rab3A antibody [EPR26022-9] (ab302518)

  Immunohistochemical analysis of paraffin-embedded Human skeletal muscl tissue labeling Rab3A with ab302518 at 1/5000 (0.11 ug/ml) followed by a LeicaDS9800 (Bond™ Polymer Refine Detection) was used at Ready to use dilution. Negative control: No staining on human skeletal muscle. The section was incubated with ab302518 for 10 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 LeicaDS9800 (Bond™ Polymer Refine Detection) was used at Ready to use dilution. Heat mediated antigen retrieval with Citrate buffer (pH 6.0, epitope retrieval solution 1) for 20 mins

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  Immunohistochemistry - Anti-Rab3A antibody [EPR26022-9] (ab302518)

  Immunohistochemical analysis of paraffin-embedded Rat cerebellum tissue labeling Rab3A with ab302518 at 1/5000 (0.11 ug/ml) followed by a LeicaDS9800 (Bond™ Polymer Refine Detection) was used at Ready to use dilution. Cytoplasmic staining on rat cerebellum (PMID: 12937130).The section was incubated with ab302518 for 10 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 LeicaDS9800 (Bond™ Polymer Refine Detection) was used at Ready to use dilution. Heat mediated antigen retrieval with Citrate buffer (pH 6.0, epitope retrieval solution 1) for 20 mins

• 

  Immunohistochemistry - Anti-Rab3A antibody [EPR26022-9] (ab302518)

  Immunohistochemical analysis of paraffin-embedded Mouse cerebellum tissue labeling Rab3A with ab302518 at 1/5000 (0.11 ug/ml) followed by a LeicaDS9800 (Bond™ Polymer Refine Detection) was used at Ready to use dilution. Cytoplasmic staining on mouse cerebellum (PMID: 10407024).The section was incubated with ab302518 for 10 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 LeicaDS9800 (Bond™ Polymer Refine Detection) was used at Ready to use dilution. Heat mediated antigen retrieval with Citrate buffer (pH 6.0, epitope retrieval solution 1) for 20 mins

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  Immunohistochemistry - Anti-Rab3A antibody [EPR26022-9] (ab302518)

  Immunohistochemical analysis of paraffin-embedded Human cerebrum tissue labeling Rab3A with ab302518 at 1/5000 (0.11 ug/ml) followed by a LeicaDS9800 (Bond™ Polymer Refine Detection) was used at Ready to use dilution. Cytoplasmic staining on human cerebrum (PMID: 26076492).The section was incubated with ab302518 for 10 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 LeicaDS9800 (Bond™ Polymer Refine Detection) was used at Ready to use dilution. Heat mediated antigen retrieval with Citrate buffer (pH 6.0, epitope retrieval solution 1) for 20 mins

• 

  Immunohistochemistry - Anti-Rab3A antibody [EPR26022-9] (ab302518)

  Immunohistochemical analysis of paraffin-embedded Human cerebellum tissue labeling Rab3A with ab302518 at 1/5000 (0.11 ug/ml) followed by a LeicaDS9800 (Bond™ Polymer Refine Detection) was used at Ready to use dilution. Cytoplasmic staining on human cerebellum. The section was incubated with ab302518 for 10 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 LeicaDS9800 (Bond™ Polymer Refine Detection) was used at Ready to use dilution. Heat mediated antigen retrieval with Citrate buffer (pH 6.0, epitope retrieval solution 1) for 20 mins