Anti-VAMP4 antibody [EPR25066-77]

• Flow Cyt (Intra)

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Flow Cytometry (Intracellular) - Anti-VAMP4 antibody [EPR25066-77] (AB290726)

Intracellular flow cytometric analysis of 4% paraformaldehyde fixed 90% methanol permeabilized 293T (Human embryonic kidney epithelial cell) cells labelling VAMP4 with ab290726 at 1/50 dilution (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 Anti-Rabbit IgG (Alexa Fluor^® 488, ab150081) at 1/2000 dilution was used as the secondary antibody.

• IHC-Fr

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Immunohistochemistry (Frozen sections) - Anti-VAMP4 antibody [EPR25066-77] (AB290726)

Immunohistochemical analysis of 4% PFA-fixed, 0.2% Triton X-100 permeabilized frozen Mouse skeletal muscle (fresh) tissue labeling VAMP4 with ab290726 at 1/500 (1.102 ug/ml) dilution followed by ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor^® 488) preadsorbed at 1/1000 (2 μg/mL) dilution (Green). Negative control (PMID : 10359608) No staining on mouse skeletal muscle is observed. The nuclear counterstain was DAPI (Blue).

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

• IHC-P

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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-VAMP4 antibody [EPR25066-77] (AB290726)

Immunohistochemical analysis of paraffin-embedded Mouse cerebrum tissue labelling VAMP4 with ab290726 at 1/500 (1.102 ug/ml) followed by a ready to use LeicaDS9800 (BOND^TM Polymer Refine Detection). Cytoplasmic staining on mouse cerebrum (PMID : 16506193). The section was incubated with ab290726 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^TM Polymer Refine Detection) was used.

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

• IHC-Fr

Supplier Data

Immunohistochemistry (Frozen sections) - Anti-VAMP4 antibody [EPR25066-77] (AB290726)

Immunohistochemical analysis of 4% PFA-fixed, 0.2% Triton X-100 permeabilized frozen Rat testis (fresh) tissue labeling VAMP4 with ab290726 at 1/500 (1.102 ug/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 testis 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.

• IHC-Fr

Supplier Data

Immunohistochemistry (Frozen sections) - Anti-VAMP4 antibody [EPR25066-77] (AB290726)

Immunohistochemical analysis of 4% PFA-fixed, 0.2% Triton X-100 permeabilized frozen Mouse testis (fresh) tissue labeling VAMP4 with ab290726 at 1/500 (1.102 ug/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 testis is observed. The nuclear counterstain was DAPI (Blue).

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

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-VAMP4 antibody [EPR25066-77] (AB290726)

Immunohistochemical analysis of paraffin-embedded Rat testis tissue labelling VAMP4 with ab290726 at 1/500 (1.102 ug/ml) followed by a ready to use LeicaDS9800 (BOND^TM Polymer Refine Detection). Cytoplasmic staining on rat testis. The section was incubated with ab290726 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^TM Polymer Refine Detection).

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

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-VAMP4 antibody [EPR25066-77] (AB290726)

Immunohistochemical analysis of paraffin-embedded Mouse testis tissue labelling VAMP4 with ab290726 at 1/500 (1.102 ug/ml) followed by a ready to use LeicaDS9800 (BOND^TM Polymer Refine Detection). Cytoplasmic staining on mouse testis. The section was incubated with ab290726 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^TM Polymer Refine Detection) was used.

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

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-VAMP4 antibody [EPR25066-77] (AB290726)

Immunohistochemical analysis of paraffin-embedded Rat cerebrum tissue labelling VAMP4 with ab290726 at 1/500 (1.102 ug/ml) followed by a ready to use LeicaDS9800 (BOND^TM Polymer Refine Detection) was used. Cytoplasmic staining on rat cerebrum. The section was incubated with ab290726 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^TM Polymer Refine Detection).

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

• ICC/IF

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Immunocytochemistry/ Immunofluorescence - Anti-VAMP4 antibody [EPR25066-77] (AB290726)

Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% TritonX-100 permeilized RAW 264.7 (mouse Abelson murine leukemia virus-induced tumor macrophage) cells lebelling VAMP4 with ab290726 at 1/50 (11.02 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 RAW 264.7 cell line is observed. 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.

• Flow Cyt (Intra)

Supplier Data

Flow Cytometry (Intracellular) - Anti-VAMP4 antibody [EPR25066-77] (AB290726)

Intracellular flow cytometric analysis of 4% paraformaldehyde fixed 90% methanol permeabilized RAW 264.7 (Mouse Abelson murine leukemia virus-induced tumor macrophage) cells labelling VAMP4 with ab290726 at 1/50 dilution (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 Anti-Rabbit IgG (Alexa Fluor^® 488, ab150081) at 1/2000 dilution was used as the secondary antibody.

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-VAMP4 antibody [EPR25066-77] (AB290726)

Immunohistochemical analysis of paraffin-embedded Human cerebrum tissue labelling VAMP4 with ab290726 at 1/500 (1.102 ug/ml) followed by a ready to use LeicaDS9800 (BOND^TM Polymer Refine Detection). Cytoplasmic staining on human cerebrum.The section was incubated with ab290726 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^TM Polymer Refine Detection) was used.

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

• IHC-Fr

Supplier Data

Immunohistochemistry (Frozen sections) - Anti-VAMP4 antibody [EPR25066-77] (AB290726)

Immunohistochemical analysis of 4% PFA-fixed, 0.2% Triton X-100 permeabilized frozen Rat skeletal muscle (fresh) tissue labeling VAMP4 with ab290726 at 1/500 (1.102 ug/ml) dilution followed by ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor^® 488) preadsorbed at 1/1000 (2 μg/mL) dilution (Green). Negative control (PMID : 10359608). No staining on rat skeletal muscle is observed. The nuclear counterstain was DAPI (Blue).

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

• IP

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Immunoprecipitation - Anti-VAMP4 antibody [EPR25066-77] (AB290726)

VAMP4 was immunoprecipitated from 0.35 mg 293T (human embryonic kidney epithelial cell) whole cell lysate 10 ug with ab290726 at 1/30 dilution (2ug in 0.35mg lysates). Western blot was performed on the immunoprecipitate using ab290726 at 1/1000 dilution. VeriBlot for IP secondary antibody(HRP)(ab131366) was used at 1/5000 dilution.

Lane 1 : 293T (human embryonic kidney epithelial cell) whole cell lysate 10 ug

Lane 2 : ab290726 IP in 293T whole cell lysate

Lane 3 : Rabbit monoclonal IgG (ab172730) instead of ab290726 in 293T whole cell lysate

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

Exposure time : 93 seconds

All lanes:

Immunoprecipitation - Anti-VAMP4 antibody [EPR25066-77] (ab290726)

Predicted band size: 16 kDa

false

• IP

Supplier Data

Immunoprecipitation - Anti-VAMP4 antibody [EPR25066-77] (AB290726)

VAMP4 was immunoprecipitated from 0.35 mg RAW264.7 (mouse Abelson murine leukemia virus-induced tumor macrophage) whole cell lysate 10 ug with ab290726 at 1/30 dilution (2ug in 0.35mg lysates). Western blot was performed on the immunoprecipitate using ab290726 at 1/1000 dilution. VeriBlot for IP secondary antibody(HRP)(ab131366) was used at 1/5000 dilution.

Lane 1 : RAW264.7 (mouse Abelson murine leukemia virus-induced tumor macrophage) whole cell lysate 10 ug

Lane 2 : ab290726 IP in RAW264.7 whole cell lysate

Lane 3 : Rabbit monoclonal IgG (ab172730) instead of ab290726 in RAW264.7 whole cell lysate

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

Exposure time : 93 seconds

All lanes:

Immunoprecipitation - Anti-VAMP4 antibody [EPR25066-77] (ab290726)

Predicted band size: 16 kDa

false

• WB

Lab

Western blot - Anti-VAMP4 antibody [EPR25066-77] (AB290726)

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

The expression profile/molecular weight observed is consistent with that described in the literature (PMID : 10359608).

All lanes:

Western blot - Anti-VAMP4 antibody [EPR25066-77] (ab290726) at 1/1000 dilution

Lane 1:

SH-SY5Y (human neuroblastoma epithelial cell) whole cell lysate. at 20 µg

Lane 2:

Human testis tissue lysate. at 20 µg

Secondary

All lanes:

Goat Anti-Rabbit IgG (HRP) with minimal cross-reactivity with human IgG at 1/2000 dilution

Predicted band size: 16 kDa

Observed band size: 18 kDa

false

Exposure time: 3min

• WB

Lab

Western blot - Anti-VAMP4 antibody [EPR25066-77] (AB290726)

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

The expression profile/molecular weight observed is consistent with that described in the literature (PMID : 10359608).

All lanes:

Western blot - Anti-VAMP4 antibody [EPR25066-77] (ab290726) at 1/1000 dilution

Lane 1:

C6 (rat glial tumor glial cell) whole cell lysate. at 20 µg

Lane 2:

RAW264.7 (mouse Abelson murine leukemia virus-induced tumor macrophage) whole cell lysate. at 20 µg

Lane 3:

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

Lane 4:

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

Secondary

All lanes:

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

Predicted band size: 16 kDa

Observed band size: 18 kDa

false

Exposure time: 3min

• WB

Lab

Western blot - Anti-VAMP4 antibody [EPR25066-77] (AB290726)

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

The expression profile/molecular weight observed is consistent with that described in the literature (PMID : 10359608).
Negative control : skeletal muscle (PMID : 10359608)

All lanes:

Western blot - Anti-VAMP4 antibody [EPR25066-77] (ab290726) at 1/1000 dilution

Lane 1:

Mouse brain tissue lysate. at 20 µg

Lane 2:

Mouse testis tissue lysate. at 20 µg

Lane 3:

Mouse skeletal muscle tissue lysate. at 20 µg

Lane 4:

Rat brain tissue lysate. at 20 µg

Lane 5:

Rat testis tissue lysate. at 20 µg

Lane 6:

Rat skeletal muscle tissue lysate. at 20 µg

Secondary

All lanes:

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

Predicted band size: 16 kDa

Observed band size: 18 kDa

false

Exposure time: 3min

• WB

Lab

Western blot - Anti-VAMP4 antibody [EPR25066-77] (AB290726)

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

The expression profile/molecular weight observed is consistent with that described in the literature (PMID : 10359608).

All lanes:

Western blot - Anti-VAMP4 antibody [EPR25066-77] (ab290726) at 1/1000 dilution

Lane 1:

Jurkat (human T cell leukemia T lymphocyte) whole cell lysate. at 40 µg

Lane 2:

K-562 (human chronic myelogenous leukemia lymphoblast) whole cell lysate at 40 µg

Lane 3:

293T (human embryonic kidney epithelial cell) whole cell lysate. at 40 µg

Lane 4:

HeLa (human cervix adenocarcinoma epithelial cell) whole cell lysate. at 40 µg

Secondary

All lanes:

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

Predicted band size: 16 kDa

Observed band size: 18 kDa

false

Exposure time: 3min