Anti-RAB35 antibody (ab152138)
Key features and details
- Rabbit polyclonal to RAB35
- Suitable for: WB, IHC-P, ICC/IF
- Knockout validated
- Reacts with: Mouse, Human
- Isotype: IgG
Overview
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Product name
Anti-RAB35 antibody
See all RAB35 primary antibodies -
Description
Rabbit polyclonal to RAB35 -
Host species
Rabbit -
Tested applications
Suitable for: WB, IHC-P, ICC/IFmore details -
Species reactivity
Reacts with: Mouse, Human -
Immunogen
Recombinant fragment, corresponding to a region within amino acids 1-201 of Human RAB35.
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Positive control
- WB: U-87 MG, SK-N-SH, IMR32, SK-N-AS, HeLa, Jurkat, Caco-2 cell lysate; transfected HEK-293T cells and mouse brain tissue lysate; IHC-P: Human breast cancer tissue; IF: U87-MG whole cell.
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General notes
The Life Science industry has been in the grips of a reproducibility crisis for a number of years. Abcam is leading the way in addressing this with our range of recombinant monoclonal antibodies and knockout edited cell lines for gold-standard validation. Please check that this product meets your needs before purchasing.
If you have any questions, special requirements or concerns, please send us an inquiry and/or contact our Support team ahead of purchase. Recommended alternatives for this product can be found below, along with publications, customer reviews and Q&As
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 or -80°C. Avoid freeze / thaw cycle. -
Storage buffer
pH: 7.00
Preservative: 0.025% Proclin 300
Constituents: 79% PBS, 20% Glycerol (glycerin, glycerine) -
Concentration information loading...
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Purity
Immunogen affinity purified -
Clonality
Polyclonal -
Isotype
IgG -
Research areas
Associated products
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Compatible Secondaries
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Isotype control
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Recombinant Protein
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Related Products
Applications
The Abpromise guarantee
Our Abpromise guarantee covers the use of ab152138 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 |
1/500 - 1/3000. Predicted molecular weight: 23 kDa.
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IHC-P |
1/100 - 1/1000. Perform heat mediated antigen retrieval before commencing with IHC staining protocol using 10mM Citrate buffer (pH6.0) or Tris-EDTA buffer (pH8.0).
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ICC/IF |
1/100 - 1/1000.
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Notes |
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WB
1/500 - 1/3000. Predicted molecular weight: 23 kDa. |
IHC-P
1/100 - 1/1000. Perform heat mediated antigen retrieval before commencing with IHC staining protocol using 10mM Citrate buffer (pH6.0) or Tris-EDTA buffer (pH8.0). |
ICC/IF
1/100 - 1/1000. |
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
- Omim: 604199 Human
- SwissProt: Q15286 Human
- SwissProt: Q6PHN9 Mouse
- Unigene: 524788 Human
- Unigene: 27553 Mouse
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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 (ab152138) at 1/500 dilution
Lane 1 : Wild-type HeLa cell lysate
Lane 2 : RAB35 knockout HeLa cell lysate
Lane 3 : Caco-2 cell lysate
Lane 4 : Jurkat cell lysate
Performed under reducing conditions.
Predicted band size: 23 kDa
Observed band size: 25 kDa why is the actual band size different from the predicted?False colour image of Western blot: Anti-RAB35 antibody staining at 1/500 dilution, shown in green; Mouse anti-Alpha Tubulin [DM1A] (ab7291) loading control staining at 1/20000 dilution, shown in red. In Western blot, ab152138 was shown to bind specifically to RAB35. A band was observed at 25 kDa in wild-type HeLa cell lysates with no signal observed at this size in RAB35 knockout cell line ab265984 (knockout cell lysate ab259076). To generate this image, wild-type and RAB35 knockout HeLa cell lysates were analysed. First, samples were run on an SDS-PAGE gel then transferred onto a nitrocellulose membrane. Membranes were blocked in 5 % milk in TBS-0.1 % Tween® 20 (TBS-T) 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 1/20000 dilution.
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All lanes : Anti-RAB35 antibody (ab152138) at 1/3000 dilution
Lane 1 : U-87 MG (human glioblastoma-astrocytoma epithelial cell line) whole cell lysate
Lane 2 : SK-N-SH (human neuroblastoma cell line) whole cell lysate
Lane 3 : IMR32 (human brain neuroblast cell line) whole cell lysate
Lane 4 : SK-N-AS (human brain neuroblastoma cell line) whole cell lysate
Lysates/proteins at 30 µg per lane.
Secondary
All lanes : HRP-conjugated anti-rabbit IgG antibody
Predicted band size: 23 kDa12% SDS-PAGE
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Immunohistochemical analysis of paraffin-embedded human breast cancer tissue staining RAB35 with ab152138 at 1/250 dilution.
Antigen Retrieval: Citrate buffer, pH 6.0, 15 min.
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Immunofluorescence analysis of 4% paraformaldehyde-fixed U-87 MG (human glioblastoma-astrocytoma epithelial cell line) whole cell labeling RAB35 with ab152138 at 1/500 dilution. Blue: Hoechst 33342 staining.
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Anti-RAB35 antibody (ab152138) at 1/1000 dilution + Mouse brain tissue lysate at 50 µg
Secondary
HRP-conjugated anti-rabbit IgG antibody
Predicted band size: 23 kDa12% SDS-PAGE
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All lanes : Anti-RAB35 antibody (ab152138) at 1/2000 dilution
Lane 1 : Non-transfected HEK-293T (human epithelial cell line from embryonic kidney transformed with large T antigen) whole cell lysate
Lane 2 : Transfected HEK-293T whole cell lysate
Lysates/proteins at 30 µg per lane.
Secondary
All lanes : HRP-conjugated anti-rabbit IgG antibody
Predicted band size: 23 kDa12% SDS-PAGE
Protocols
Datasheets and documents
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SDS download
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Datasheet download
References (3)
ab152138 has been referenced in 3 publications.
- Wen H et al. MiR-185-5p targets RAB35 gene to regulate tumor cell-derived exosomes-mediated proliferation, migration and invasion of non-small cell lung cancer cells. Aging (Albany NY) 13:21435-21450 (2021). PubMed: 34500436
- Faris R et al. Chlamydia trachomatis CT229 Subverts Rab GTPase-Dependent CCV Trafficking Pathways to Promote Chlamydial Infection. Cell Rep 26:3380-3390.e5 (2019). PubMed: 30893609
- Corallino S et al. A RAB35-p85/PI3K axis controls oscillatory apical protrusions required for efficient chemotactic migration. Nat Commun 9:1475 (2018). PubMed: 29662076