Anti-Rab3A antibody (ab3335)
Key features and details
- Rabbit polyclonal to Rab3A
- Suitable for: WB, ICC/IF
- Knockout validated
- Reacts with: Mouse, Rat, Human
- Isotype: IgG
Overview
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Product name
Anti-Rab3A antibody
See all Rab3A primary antibodies -
Description
Rabbit polyclonal to Rab3A -
Host species
Rabbit -
Tested applications
Suitable for: WB, ICC/IFmore details -
Species reactivity
Reacts with: Mouse, Rat, Human -
Immunogen
Synthetic peptide corresponding to Human Rab3A aa 1-18.
Sequence:MASATDSRYGQKESSDQN
(Peptide available asab4951) -
Positive control
- WB: mouse brain, kidney, liver, rat brain, kidney, liver; ICC: HeLa cells
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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
Preservative: 0.05% Sodium azide
Constituent: 0.1% BSA -
Concentration information loading...
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Purity
Protein A purified -
Primary antibody notes
Rab proteins are low-molecular-weight GTP-binding proteins that form the largest branch of the Ras superfamily of GTPases. Located on the cytoplasmic face of organelles and vesicles, rab proteins are involved in intracellular membrane fusion reactions. Three membrane proteins, synaptosomal associated protein of 25 kDa (SNAP-25), synaptobrevin and syntaxin, form the core of a ubiquitous membrane fusion machine that interacts with the soluble proteins N-ethylmaleimide-sensitive factor (NSF) and a-SNAP. Rab proteins, in co-ordination with the core fusion machinery and Munc-18, help to mediate vesicle docking and fusion. There exist over 40 rab proteins that have been described in mammals and the best studied is rab 3A. Rab 3A is found to be abundant in presynaptic nerve terminals and is also found to be crucial in acrosomal exocytosis in human spermatozoa. Abnormal accumulation of rab 3A in the cytoplasm of Purkinje cells has been reported in the prion protein related Creutzfeldt-Jakob disease. -
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
Applications
The Abpromise guarantee
Our Abpromise guarantee covers the use of ab3335 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 | (7) |
Use a concentration of 2 µg/ml.
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ICC/IF | (1) |
Use a concentration of 2 µg/ml.
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Notes |
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WB
Use a concentration of 2 µg/ml. |
ICC/IF
Use a concentration of 2 µg/ml. |
Target
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Function
Involved in exocytosis by regulating a late step in synaptic vesicle fusion. Could play a role in neurotransmitter release by regulating membrane flow in the nerve terminal. -
Tissue specificity
Specifically expressed in brain. -
Sequence similarities
Belongs to the small GTPase superfamily. Rab family. -
Cellular localization
Cell membrane. - Information by UniProt
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Database links
- Entrez Gene: 5864 Human
- Entrez Gene: 19339 Mouse
- Entrez Gene: 25531 Rat
- Omim: 179490 Human
- SwissProt: P20336 Human
- SwissProt: P63011 Mouse
- SwissProt: P63012 Rat
- Unigene: 27744 Human
see all -
Alternative names
- Rab 3A antibody
- RAB 3A member RAS oncogene family antibody
- Rab3a antibody
see all
Images
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All lanes : Left: ab3335 at 2 µg/mL.
Right: ab302518 at 1/1000 dilution
Lane 1 : Wild-type SK-N-FI cell lysate
Lane 2 : RAB3A knockout SK-N-FI cell lysate
Lane 3 : Human brain cell lysate
Lane 4 : HEK-293 cell lysate
Lysates/proteins at 20 µg per lane.
Developed using the ECL technique.
Performed under reducing conditions.
Observed band size: 27 kDa why is the actual band size different from the predicted?Western blot: Left: Anti-RAB3A antibody (ab3335) staining at 2 ug/ml. Right: Recombinant Anti-Rab3A antibody [EPR26022-9] (ab302518) staining at 1/1000 dilution. Anti-Rab3A antibodies shown in green; Mouse anti-Alpha Tubulin [DM1A] (ab7291) loading control staining at 1/20000 dilution, shown in magenta. In Western blot, ab3335 and ab302518 were shown to bind specifically to RAB3A. A band was observed at 27 kDa in wild-type SK-N-FI cell lysates with no signal observed at this size in RAB3A knockout cell line. To generate this image, wild-type and RAB3A knockout SK-N-FI cell lysates were analysed. First, samples were run on an SDS-PAGE gel then transferred onto a nitrocellulose membrane. Membranes were blocked in 3 % 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 800CW and Goat anti-Mouse IgG H&L 680RD at 1/20000 dilution.
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Immunofluorescence analysis of RAB3A was done on 70% confluent log phase HeLa (Human epithelial adenocarcinoma cell line)
cells. The cells were fixed with 4% paraformaldehyde for 10 minutes, permeabilized with 0.1% Triton™ X-100 for 10 minutes, and blocked with 1% BSA for 1 hour at room temperature. The cells were labeled with ab3335 at 2 µg/mL in 0.1% BSA and incubated for 3 hours at room temperature and then labeled with Goat anti-Rabbit IgG (H+L) Superclonal™ Secondary Antibody, Alexa Fluor® 488 conjugate at 1/2000 dilution for 45 minutes at room temperature (Panel a: green). Nuclei (Panel b: blue) were stained with SlowFade® Gold Antifade Mountant with DAPI. F-actin (Panel c: red) was stained with Alexa Fluor® 555 Rhodamine Phalloidin at 1/300 dilution. Panel d is a merged image showing Cytoplasmic localization. Panel e is a no primary antibody control. The images were captured at 60X magnification. -
All lanes : Anti-Rab3A antibody (ab3335) at 2 µg/ml
Lane 1 : Mouse brain tissue lysate
Lane 2 : Mouse kidney tissue lysate
Lane 3 : Mouse liver tissue lysate
Lane 4 : Rat brain tissue lysate
Lane 5 : Rat kidney tissue lysate
Lane 6 : Rat liver tissue lysate
Lysates/proteins at 30 µg per lane.
Secondary
All lanes : Rabbit anti-Goat IgG (H+L) Superclonal™ Recombinant Secondary Antibody, HRP at 1/4000 dilutionA 27 kDa band corresponding to RAB3A was observed along with two uncharacterized band (*) at ~40 kD and 60 kDa across tissues tested.
Protocols
Datasheets and documents
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SDS download
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Datasheet download
References (16)
ab3335 has been referenced in 16 publications.
- Astillero-Lopez V et al. Neurodegeneration and astrogliosis in the entorhinal cortex in Alzheimer's disease: Stereological layer-specific assessment and proteomic analysis. Alzheimers Dement 18:2468-2480 (2022). PubMed: 35142030
- Prasai B et al. The nanoscale molecular morphology of docked exocytic dense-core vesicles in neuroendocrine cells. Nat Commun 12:3970 (2021). PubMed: 34172739
- Wauters F et al. LRRK2 mutations impair depolarization-induced mitophagy through inhibition of mitochondrial accumulation of RAB10. Autophagy 16:203-222 (2020). PubMed: 30945962
- Connor-Robson N et al. An integrated transcriptomics and proteomics analysis reveals functional endocytic dysregulation caused by mutations in LRRK2. Neurobiol Dis 127:512-526 (2019). PubMed: 30954703
- Folsom TD et al. Quantitative proteomics of forebrain subcellular fractions in fragile X mental retardation 1 knockout mice following acute treatment with 2-Methyl-6-(phenylethynyl)pyridine: Relevance to developmental study of schizophrenia. Synapse 73:e22069 (2019). PubMed: 30176067