Recombinant Anti-RAB35 antibody [EPR24487-21] (ab300116)
![Western blot - Anti-RAB35 antibody [EPR24487-21] (AB300116)](https://www.abcam.com/ps/products/300/ab300116/Images/AB300116-485982-WB2.jpg "Western blot - Anti-RAB35 antibody [EPR24487-21] (AB300116)")
Key features and details
- Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
- Rabbit monoclonal [EPR24487-21] to RAB35
- Suitable for: WB, IP
- Knockout validated
- Reacts with: Mouse, Rat, Human
Related conjugates and formulations
Overview
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Product name
Anti-RAB35 antibody [EPR24487-21]
See all RAB35 primary antibodies -
Description
Rabbit monoclonal [EPR24487-21] to RAB35 -
Host species
Rabbit -
Tested applications
Suitable for: WB, IPmore details
Unsuitable for: Flow Cyt (Intra),ICC/IF or IHC-P -
Species reactivity
Reacts with: Mouse, Rat, Human -
Immunogen
Recombinant fragment. This information is proprietary to Abcam and/or its suppliers.
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Positive control
- WB: Rat and mouse brain, Caco-2, 293T, C6, RAW264.7, PC-12, NIH/3T3, RAB35 full length protein lysates. IP: Caco-2 cell.
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General notes
Ab300116 does not reactin in IP with Mouse species.
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.
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... -
Purity
Protein A purified -
Clonality
Monoclonal -
Clone number
EPR24487-21 -
Isotype
IgG -
Research areas
Associated products
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Alternative Versions
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Compatible Secondaries
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Isotype control
Applications
The Abpromise guarantee
Our Abpromise guarantee covers the use of ab300116 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 |
|---|---|---|
| WB |
1/1000. Detects a band of approximately 25 kDa (predicted molecular weight: 23 kDa).
|
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| IP |
1/30.
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| Notes |
|---|
|
WB
1/1000. Detects a band of approximately 25 kDa (predicted molecular weight: 23 kDa). |
|
IP
1/30. |
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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Western blot - Anti-RAB35 antibody [EPR24487-21] (AB300116)All lanes : Anti-RAB35 antibody [EPR24487-21] (ab300116) at 1/1000 dilution
Lane 1 : Rat brain tissue lysate
Lane 2 : Mouse brain tissue lysate
Lane 3 : Caco-2 (human colorectal adenocarcinoma epithelial cell) whole cell lysate
Lane 4 : 293T (human embryonic kidney epithelial cell) 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: 23 kDa
Observed band size: 25 kDa why is the actual band size different from the predicted?
Exposure time: 3 minutesBlocking and dilution buffer and concentration: 5% NFDM/TBST.
The expression profile/ molecular weight observed is consistent with what has been described in the literature (PMID: 29662076; PMID: 34349123 ).
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![Western blot - Anti-RAB35 antibody [EPR24487-21] (AB300116)](https://www.abcam.com/ps/products/300/ab300116/Images/AB300116-485979-WB1.jpg)
Western blot - Anti-RAB35 antibody [EPR24487-21] (AB300116)All lanes : Anti-RAB35 antibody [EPR24487-21] (ab300116) 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 1/20 dilution per lane.
Secondary
Lanes 1-3 : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/100000 dilution
Lane 4 : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/100000 dilution (NIH/3T3 (mouse embryonic fibroblast) whole cell lysate)
Predicted band size: 23 kDa
Observed band size: 25 kDa why is the actual band size different from the predicted?
Exposure time: 3 minutesBlocking and dilution buffer and concentration: 5% NFDM/TBST.
The expression profile/ molecular weight observed is consistent with what has been described in the literature (PMID: 23572513 ).
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![Western blot - Anti-RAB35 antibody [EPR24487-21] (AB300116)](https://www.abcam.com/ps/products/300/ab300116/Images/AB300116-485976-WB3.jpg)
Western blot - Anti-RAB35 antibody [EPR24487-21] (AB300116)All lanes : Anti-RAB35 antibody [EPR24487-21] (ab300116) 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: 26 secondsBlocking and dilution buffer and concentration: 5% NFDM/TBST.
This antibody has no cross-reactivity with other RAB family proteins.
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![Immunoprecipitation - Anti-RAB35 antibody [EPR24487-21] (AB300116)](https://www.abcam.com/ps/products/300/ab300116/Images/AB300116-2-anti-rab35-antibody-epr2448721-immunoprecipitation-caco2.jpg)
Immunoprecipitation - Anti-RAB35 antibody [EPR24487-21] (AB300116)RAB35 was immunoprecipitated from 0.35 mg Caco-2 (human colorectal adenocarcinoma epithelial cell) whole cell lysate 10 µg with ab300116 at 1/30 dilution (2µg in 0.35mg lysates). Western blot was performed on the immunoprecipitate using ab300116 at 1/1000 dilution. VeriBlot for IP secondary antibody(HRP)(ab131366) was used at 1/5000 dilution.
Lane 1 (Input): Caco-2 (human colorectal adenocarcinoma epithelial cell) whole cell lysate 10 µg
Lane 2 (+): Caco-2 whole cell lysate
Lane 3 (-): Rabbit monoclonal IgG (ab172730) instead of ab300116 in Caco-2 whole cell lysate
Blocking and dilution buffer and concentration: 5% NFDM/TBST.
Exposure time: 32 seconds
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![Anti-RAB35 antibody [EPR24487-21] (ab300116)](https://www.abcam.com/ps/products/300/ab300116/Images/AB300116-1-Benefits-of-Recombinant-antibodies.jpg)
Anti-RAB35 antibody [EPR24487-21] (ab300116)
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)
ab300116 has not yet been referenced specifically in any publications.