Recombinant Anti-RAB35 antibody [MJF-R34-25] (ab288567)
Key features and details
- Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
- Rabbit monoclonal [MJF-R34-25] to RAB35
- Suitable for: WB
- Knockout validated
- Reacts with: Mouse, Rat, Human
Related conjugates and formulations
Overview
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Product name
Anti-RAB35 antibody [MJF-R34-25]
See all RAB35 primary antibodies -
Description
Rabbit monoclonal [MJF-R34-25] to RAB35 -
Host species
Rabbit -
Tested applications
Suitable for: WBmore details
Unsuitable for: IHC-P or IP -
Species reactivity
Reacts with: Mouse, Rat, Human -
Immunogen
Recombinant full length protein. This information is proprietary to Abcam and/or its suppliers.
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Positive control
- WB: Wild-type A549, Caco-2, Jurkat, 293T and MEF whole cell lysates, and Mouse lung, Mouse brain, Rat brain tissues, and RAB35 full-length protein.
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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
Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMAb® patents.
This antibody was developed with support from The Michael J. Fox Foundation.
Properties
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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...
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Purity
Protein A purified -
Clonality
Monoclonal -
Clone number
MJF-R34-25 -
Isotype
IgG -
Research areas
Associated products
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Compatible Secondaries
Applications
The Abpromise guarantee
Our Abpromise guarantee covers the use of ab288567 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 |
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WB |
1/1000. Predicted molecular weight: 23 kDa.
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Notes |
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WB
1/1000. Predicted molecular weight: 23 kDa. |
Target
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Function
In the process of endocytosis, essential rate-limiting regulator of a fast recycling pathway back to the plasma membrane. During cytokinesis, required for the postfurrowing terminal steps, namely for intercellular bridge stability and abscission, possibly by controlling phosphatidylinositol 4,5-bis phosphate (PIP2) and SEPT2 localization at the intercellular bridge. -
Sequence similarities
Belongs to the small GTPase superfamily. Rab family. -
Cellular localization
Cell membrane. Membrane > clathrin-coated pit. Cytoplasmic vesicle > clathrin-coated vesicle. Endosome. Melanosome. Present on sorting endosomes and endosomal tubules. Tends to be enriched in PIP2-positive cell membrane domains. During mitosis, associated with the plasma membrane and present at the ingressing furrow during early cytokinesis as well as at the intercellular bridge later during cytokinesis. Identified by mass spectrometry in melanosome fractions from stage I to stage IV. - Information by UniProt
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Database links
- Entrez Gene: 11021 Human
- Entrez Gene: 77407 Mouse
- Entrez Gene: 288700 Rat
- Omim: 604199 Human
- SwissProt: Q15286 Human
- SwissProt: Q6PHN9 Mouse
- SwissProt: Q5U316 Rat
- Unigene: 524788 Human
see all -
Alternative names
- GTP binding protein RAY antibody
- GTP-binding protein RAY antibody
- H ray antibody
see all
Images
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All lanes : Anti-RAB35 antibody [MJF-R34-25] (ab288567) at 1/1000 dilution
Lane 1 : Wild-type A549 (human lung carcinoma epithelial cell) whole cell lysate
Lane 2 : RAB35 knockout A549 whole cell lysate
Lane 3 : Caco-2 (human colorectal adenocarcinoma epithelial cell) whole cell lysate
Lane 4 : Jurkat (human T cell leukemia T lymphocyte) whole cell lysate
Secondary
All lanes : Goat Anti-Rabbit IgG H&L (IRDye® 800CW) (ab216773) and Goat Anti-Mouse IgG H&L (IRDye® 680RD) (ab216776) at 1/10000 dilution
Predicted band size: 23 kDa
Observed band size: 25 kDa why is the actual band size different from the predicted?Blocking and diluting buffer and concentration: Intercept® (TBS) Blocking Buffer diluted with an equal volume of 0.1% TBS
Lysates/proteins at 20 µg per lane.
Performed under reducing conditions.
False colour image of Western blot: Anti-RAB35 antibody [MJF-34-R25] (ab288567) staining at 1/1000 dilution, shown in green; Mouse anti-GAPDH antibody [6C5] (ab8245) loading control staining at 1/20000 dilution, shown in red.
In Western blot, ab288567 was shown to bind specifically to RAB35. A band was observed at 25 kDa in wild-type A549 cell lysates with no signal observed at this size in RAB35 knockout cell lysates. To generate this image, wild-type and RAB35 knockout A549 cell lysates were analyzed. First, samples were run on an SDS-PAGE gel then transferred onto an immobilon-FL PVDF membrane. Membranes were blocked in Intercept® (TBS) Blocking Buffer diluted with an equal volume of 0.1% TBS 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 (IRDye® 800CW) preabsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye® 680RD) preabsorbed (ab216776) at 1/10000 dilution.
Wild-type A549 and RAB35 knockout A549 lysates were kindly provided by Prof. Dario Alessi University of Dundee.
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All lanes : Anti-RAB35 antibody [MJF-R34-25] (ab288567) at 1/1000 dilution
Lane 1 : 293T (human embryonic kidney epithelial cell) whole cell lysate
Lane 2 : MEF (mouse embryonic fibroblast (immortalized)) whole cell lysate
Lane 3 : Mouse lung tissue lysate
Lane 4 : Mouse brain tissue lysate
Lane 5 : Rat brain 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: 23 kDa
Observed band size: 25 kDa why is the actual band size different from the predicted?
Exposure time: 37 secondsBlocking and diluting buffer and concentration: 5% NFDM/TBST
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All lanes : Anti-RAB35 antibody [MJF-R34-25] (ab288567) at 1/1000 dilution
Lane 1 : RAB35 full-length protein 10 ng
Lane 2 : RAB family protein homologous region 10 ng
Secondary
All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/100000 dilution
Predicted band size: 23 kDa
Observed band size: 25 kDa why is the actual band size different from the predicted?
Exposure time: 15 secondsBlocking and diluting buffer and concentration: 5% NFDM/TBST
This antibody has no cross-reactivity with other RAB family proteins
Protocols
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.
Datasheets and documents
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SDS download
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Datasheet download
Certificate of Compliance
References (0)
ab288567 has not yet been referenced specifically in any publications.