Anti-Cellubrevin antibody [EPR25065-16]

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Cellubrevin antibody [EPR25065-16] (AB283312)

Immunohistochemical analysis of paraffin-embedded Mouse kidney tissue labelling Cellubrevin with ab283312 at 1/2000 (0.287 ug/ml) followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection). Positive staining in mouse kidney. The section was incubated with ab283312 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 LeicaDS9800 (Bond™ Polymer Refine Detection).

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

• Flow Cyt (Intra)

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Flow Cytometry (Intracellular) - Anti-Cellubrevin antibody [EPR25065-16] (AB283312)

Flow cytometric analysis of 4% paraformaldehyde fixed 90% methanol permeabilized NIH/3T3 (Mouse embryonic fibroblast) cells labelling Cellubrevin with ab283312 at 1/500 dilution (0.1ug) (Red) compared with a Rabbit monoclonal IgG (ab172730) (Black) isotype control and an unlabelled control (cells without incubation with primary antibody and secondary antibody) (Blue). A Goat F(ab')2 Anti-Rabbit IgG (DyLight® 488, ab98507) at 1/500 dilution was used as the secondary antibody.

• ICC/IF

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Immunocytochemistry/ Immunofluorescence - Anti-Cellubrevin antibody [EPR25065-16] (AB283312)

Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% TritonX-100 permeabilized Neuro-2a cells labelling Cellubrevin with ab283312 at 1/500 (11.46 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 cytoplasmic staining in Neuro-2a cell line. ab195889 Anti-alpha Tubulin mouse monoclonal antibody - Microtubule Marker (Alexa Fluor® 594) was used to counterstain tubulin at 1/200 (2.5 ug/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

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Cellubrevin antibody [EPR25065-16] (AB283312)

Immunohistochemical analysis of paraffin-embedded Mouse testis tissue labelling Cellubrevin with ab283312 at 1/2000 (0.287 ug/ml) followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection). Positive staining in mouse testis. The section was incubated with ab283312 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 LeicaDS9800 (Bond™ Polymer Refine Detection).

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

• Flow Cyt (Intra)

Supplier Data

Flow Cytometry (Intracellular) - Anti-Cellubrevin antibody [EPR25065-16] (AB283312)

Flow cytometric analysis of 4% paraformaldehyde fixed 90% methanol permeabilized PC-12 (Rat adrenal gland pheochromocytoma) cells labelling Cellubrevin with ab283312 at 1/500 dilution (0.1ug) (Red) compared with a Rabbit monoclonal IgG (ab172730) (Black) isotype control and an unlabelled control (cells without incubation with primary antibody and secondary antibody) (Blue). A Goat F(ab')2 Anti-Rabbit IgG (DyLight® 488, ab98507) at 1/500 dilution was used as the secondary antibody.

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Cellubrevin antibody [EPR25065-16] (AB283312)

Immunohistochemical analysis of paraffin-embedded Rat testis tissue labelling Cellubrevin with ab283312 at 1/5000 (0.1146 ug/ml) followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection). Cytoplasmic staining in rat testis. The section was incubated with ab283312 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 LeicaDS9800 (Bond™ Polymer Refine Detection).

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

• IP

Supplier Data

Immunoprecipitation - Anti-Cellubrevin antibody [EPR25065-16] (AB283312)

Cellubrevin was immunoprecipitated from 0.35 mg Rat testis tissue lysate 10ug with ab283312 at 1/30 dilution (2ug in 0.35mg lysates). Western blot was performed on the immunoprecipitate using ab283312 at 1/1000 dilution. VeriBlot for IP secondary antibody(HRP)(ab131366) was used at 1/5000 dilution.

Lane 1 : Rat testis tissue lysate 10ug

Lane 2 : ab283312 IP in Rat testis tissue lysate

Lane 3 : Rabbit monoclonal IgG (ab172730) instead of ab283312 in rat testis tissue lysate

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

Exposure time : 3 seconds.

All lanes:

Immunoprecipitation - Anti-Cellubrevin antibody [EPR25065-16] (ab283312)

Predicted band size: 11 kDa

Observed band size: 15 kDa

false

• IP

Supplier Data

Immunoprecipitation - Anti-Cellubrevin antibody [EPR25065-16] (AB283312)

Cellubrevin was immunoprecipitated from 0.35 mg Mouse testis tissue lysate 10ug with ab283312 at 1/30 dilution (2ug in 0.35mg lysates). Western blot was performed on the immunoprecipitate using ab283312 at 1/1000 dilution. VeriBlot for IP secondary antibody(HRP)(ab131366) was used at 1/5000 dilution.

Lane 1 : Mouse testis tissue lysate 10ug

Lane 2 : ab283312 IP in Mouse testis tissue lysate

Lane 3 : Rabbit monoclonal IgG (ab172730) instead of ab283312 in mouse testis tissue lysate

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

Exposure time : 5 seconds.

All lanes:

Immunoprecipitation - Anti-Cellubrevin antibody [EPR25065-16] (ab283312)

Predicted band size: 11 kDa

Observed band size: 15 kDa

false

• WB

Lab

Western blot - Anti-Cellubrevin antibody [EPR25065-16] (AB283312)

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

The blot of lane 1 & 2 was developed using a higher sensitivity ECL substrate.

Exposure time : Lane 1&2 : 3 minutes;

Lane 3 : 103 seconds.

All lanes:

Western blot - Anti-Cellubrevin antibody [EPR25065-16] (ab283312) at 1/1000 dilution

Lane 1:

NIH/3T3 (mouse embryonic fibroblast) whole cell lysate at 20 µg

Lane 2:

Neuro-2a (mouse neuroblastoma neuroblast) whole cell lysate at 20 µg

Lane 3:

4T1 (mouse mammary gland carcinoma epithelial cell) whole cell lysate at 20 µg

Secondary

All lanes:

Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated (<a href='/en-us/products/secondary-antibodies/goat-rabbit-igg-h-l-hrp-ab97051'>ab97051</a>) at 1/100000 dilution

Predicted band size: 11 kDa

Observed band size: 15 kDa

false

• WB

Lab

Western blot - Anti-Cellubrevin antibody [EPR25065-16] (AB283312)

Blocking and diluting buffer and concentration : 5% NFDM/TBSTLow expression : rat liver (PMID : 8332193)

Exposure time : 103 seconds

All lanes:

Western blot - Anti-Cellubrevin antibody [EPR25065-16] (ab283312) at 1/1000 dilution

Lane 1:

Rat testis tissue lysate at 20 µg

Lane 2:

Rat liver tissue lysate at 40 µg

Lane 3:

Rat skeletal muscle tissue lysate at 20 µg

Lane 4:

PC-12 (rat adrenal gland pheochromocytoma), whole cell lysate at 20 µg

Secondary

All lanes:

Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated (<a href='/en-us/products/secondary-antibodies/goat-rabbit-igg-h-l-hrp-ab97051'>ab97051</a>) at 1/100000 dilution

Predicted band size: 11 kDa

Observed band size: 15 kDa

false

• WB

Lab

Western blot - Anti-Cellubrevin antibody [EPR25065-16] (AB283312)

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

The expression profile observed is consistent with what has been described in the literature (PMID : 8332193)

Exposure time : 103 seconds.

All lanes:

Western blot - Anti-Cellubrevin antibody [EPR25065-16] (ab283312) at 1/1000 dilution

Lane 1:

Mouse testis tissue lysate at 20 µg

Lane 2:

Mouse liver tissue lysate at 40 µg

Lane 3:

Mouse skeletal muscle tissue lysate at 20 µg

Secondary

All lanes:

Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated (<a href='/en-us/products/secondary-antibodies/goat-rabbit-igg-h-l-hrp-ab97051'>ab97051</a>) at 1/100000 dilution

Predicted band size: 11 kDa

Observed band size: 15 kDa

false