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

Product name

Anti-VAMP4 antibody [EPR25066-77]
See all VAMP4 primary antibodies
Description

Rabbit monoclonal [EPR25066-77] to VAMP4
Host species

Rabbit
Specificity

Please note this antibody does not react with Human species for ICC application.
Tested applications

Suitable for: IHC-P, Flow Cyt (Intra), ICC, IHC-Fr, WB, IPmore details
Species reactivity

Reacts with: Mouse, Rat, Human
Immunogen

Recombinant fragment. This information is proprietary to Abcam and/or its suppliers.
Positive control
- WB: SH-SY5Y, Jurkat, K-562, 293T, HeLa, C6, RAW264.7, PC-12, NIH/3T3 whole cell lysates. Human testis, Mouse brain, Mouse testis, Rat brain, Rat testis whole tissue lysates. IHC-P: human cerebrum, mouse cerebrum, mouse testis, rat cerebrum, rat testis tissues. ICC: RAW 264.7. IHC-Fr: mouse testis, rat testis. Flow cyt (Intra): RAW 264.7, 293T. IP: 293T, RAW264.7
General notes
This product is a recombinant monoclonal antibody, which offers several advantages including:
- - High batch-to-batch consistency and reproducibility
- - Improved sensitivity and specificity
- - Long-term security of supply
- - Animal-free production
For more information see here.
Our RabMAb^® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMAb^® patents.

Form

Liquid
Storage instructions

Shipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C long term. Avoid freeze / thaw cycle.
Storage buffer

pH: 7.2
Preservative: 0.01% Sodium azide
Constituents: 59% PBS, 40% Glycerol (glycerin, glycerine), 0.05% BSA
Concentration information loading...
Purity

Protein A purified
Clonality

Monoclonal
Clone number

EPR25066-77
Isotype

IgG
Research areas
- Neuroscience
- Processes

The Abpromise guarantee

Our Abpromise guarantee covers the use of ab290726 in the following tested applications.

The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.

Application	Abreviews	Notes
IHC-P		1/500.
Flow Cyt (Intra)		1/50.
ICC		1/50.
IHC-Fr		1/500.
WB		1/1000. Predicted molecular weight: 16 kDa.
IP		1/30.

Notes
IHC-P 1/500.
Flow Cyt (Intra) 1/50.
ICC 1/50.
IHC-Fr 1/500.
WB 1/1000. Predicted molecular weight: 16 kDa.
IP 1/30.

Function

Involved in the pathway that functions to remove an inhibitor (probably synaptotagmin-4) of calcium-triggered exocytosis during the maturation of secretory granules. May be a marker for this sorting pathway that is critical for remodeling the secretory response of granule.
Sequence similarities

Belongs to the synaptobrevin family.
Contains 1 v-SNARE coiled-coil homology domain.
Cellular localization

Golgi apparatus > trans-Golgi network membrane. Associated with trans Golgi network (TGN) and newly formed immature secretory granules (ISG). Not found on the mature secretory organelles.
Target information above from: UniProt accession O75379 The UniProt Consortium
The Universal Protein Resource (UniProt) in 2010
Nucleic Acids Res. 38:D142-D148 (2010) .

Information by UniProt
Database links
- Entrez Gene: 8674 Human
- Entrez Gene: 53330 Mouse
- Entrez Gene: 364033 Rat
- Omim: 606909 Human
- SwissProt: A2IDD8 Human
- SwissProt: O75379 Human
- SwissProt: O70480 Mouse
- Unigene: 6651 Human
- Unigene: 10699 Mouse
see all
Alternative names
- VAMP 24 antibody
- VAMP 4 antibody
- VAMP-4 antibody
- VAMP24 antibody
- VAMP4 antibody
- VAMP4 protein antibody
- VAMP4_HUMAN antibody
- Vesicle associated membrane protein 4 antibody
- Vesicle associated membrane protein 4 isoform CRA a antibody
- Vesicle associated membrane protein 4 isoform CRA b antibody
- Vesicle-associated membrane protein 4 antibody
see all

Western blot - Anti-VAMP4 antibody [EPR25066-77] (ab290726)

All lanes : Anti-VAMP4 antibody [EPR25066-77] (ab290726) at 1/1000 dilution

Lane 1 : SH-SY5Y (human neuroblastoma epithelial cell) whole cell lysate.
Lane 2 : Human testis tissue lysate.

Lysates/proteins at 20 µg per lane.

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 why is the actual band size different from the predicted?

Exposure time: 3 minutes

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

The expression profile/molecular weight observed is consistent with that described in the literature (PMID: 10359608).
Western blot - Anti-VAMP4 antibody [EPR25066-77] (ab290726)

All lanes : Anti-VAMP4 antibody [EPR25066-77] (ab290726) at 1/1000 dilution

Lane 1 : Jurkat (human T cell leukemia T lymphocyte) whole cell lysate.
Lane 2 : K-562 (human chronic myelogenous leukemia lymphoblast) whole cell lysate
Lane 3 : 293T (human embryonic kidney epithelial cell) whole cell lysate.
Lane 4 : HeLa (human cervix adenocarcinoma epithelial cell) whole cell lysate.

Lysates/proteins at 40 µg per lane.

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

Predicted band size: 16 kDa
Observed band size: 18 kDa why is the actual band size different from the predicted?

Exposure time: 3 minutes

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

The expression profile/molecular weight observed is consistent with that described in the literature (PMID: 10359608).
Western blot - Anti-VAMP4 antibody [EPR25066-77] (ab290726)

All lanes : Anti-VAMP4 antibody [EPR25066-77] (ab290726) at 1/1000 dilution

Lane 1 : Mouse brain tissue lysate.
Lane 2 : Mouse testis tissue lysate.
Lane 3 : Mouse skeletal muscle tissue lysate.
Lane 4 : Rat brain tissue lysate.
Lane 5 : Rat testis tissue lysate.
Lane 6 : Rat skeletal muscle tissue lysate.

Lysates/proteins at 20 µg per lane.

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

Predicted band size: 16 kDa
Observed band size: 18 kDa why is the actual band size different from the predicted?

Exposure time: 3 minutes

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)
Western blot - Anti-VAMP4 antibody [EPR25066-77] (ab290726)

All lanes : Anti-VAMP4 antibody [EPR25066-77] (ab290726) at 1/1000 dilution

Lane 1 : C6 (rat glial tumor glial cell) whole cell lysate.
Lane 2 : RAW264.7 (mouse Abelson murine leukemia virus-induced tumor macrophage) whole cell lysate.
Lane 3 : PC-12 (rat adrenal gland pheochromocytoma) whole cell lysate.
Lane 4 : NIH/3T3 (mouse embryonic fibroblast) whole cell lysate.

Lysates/proteins at 20 µg per lane.

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

Predicted band size: 16 kDa
Observed band size: 18 kDa why is the actual band size different from the predicted?

Exposure time: 3 minutes

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

The expression profile/molecular weight observed is consistent with that described in the literature (PMID: 10359608).
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
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
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
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
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
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.
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.
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.
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.
Immunocytochemistry - 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 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.
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.
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
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
Anti-VAMP4 antibody [EPR25066-77] (ab290726)

To our knowledge, customised protocols are not required for this product. Please try the standard protocols listed below and let us know how you get on.

Click here to view the general protocols

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Overview

Product name

Description

Host species

Specificity

Tested applications

Species reactivity

Immunogen

Positive control

General notes

Properties

Form

Storage instructions

Storage buffer

Purity

Clonality

Clone number

Isotype

Research areas

Applications

The Abpromise guarantee

Target

Function

Sequence similarities

Cellular localization

Database links

Alternative names

Images

Protocols

Datasheets and documents

SDS download

Datasheet download

Certificate of Compliance

References (0)

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