Anti-Spastin antibody [Sp 3G11/1] (ab31850)
Key features and details
- Mouse monoclonal [Sp 3G11/1] to Spastin
- Suitable for: WB, IHC-P, IP
- Knockout validated
- Reacts with: Rat, Human
- Isotype: IgG2a
Overview
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Product name
Anti-Spastin antibody [Sp 3G11/1]
See all Spastin primary antibodies -
Description
Mouse monoclonal [Sp 3G11/1] to Spastin -
Host species
Mouse -
Tested applications
Suitable for: WB, IHC-P, IPmore details -
Species reactivity
Reacts with: Rat, Human -
Immunogen
Recombinant full length protein (Human).
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Positive control
- WB: Total HeLa extract or rat brain synaptosome This antibody gave a positive result in IHC in the following FFPE tissue: Human lung adenocarcinoma. IP: HAP1 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: 0.02% Sodium azide
Constituent: 99.98% PBS -
Concentration information loading...
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Purity
Protein A/G purified -
Clonality
Monoclonal -
Clone number
Sp 3G11/1 -
Myeloma
Sp2/0-Ag14 -
Isotype
IgG2a -
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 ab31850 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. Detects a band of approximately 52 kDa.
There are two splice isoforms of spastin, one without exon4 and two alternative ATG start sites, which may determine the localisation of the translate protein. 52kDa is the major band. |
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IHC-P |
Use at an assay dependent concentration.
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IP |
Use at an assay dependent concentration.
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Notes |
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WB
1/500. Detects a band of approximately 52 kDa. There are two splice isoforms of spastin, one without exon4 and two alternative ATG start sites, which may determine the localisation of the translate protein. 52kDa is the major band. |
IHC-P
Use at an assay dependent concentration. |
IP
Use at an assay dependent concentration. |
Target
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Relevance
Spastin is thought have a role in microtubule dynamics through its function as a microtubule severing protein. It is localised to the centrosome of neuronal cells but is not found in glial cells. Mutation in the ATPase binding domain of spastin causes hereditary spastic paraplegias (HSP), a large group of clinically similar disorders. Mutant forms of spastin are generally found throughout the cytoplasm rather then within the nucleus. -
Cellular localization
Cytoplasm, Cytoskeleton, Endoplasmic reticulum, Endosome, Membrane, Microtubule, Nucleus -
Database links
- Entrez Gene: 6683 Human
- Entrez Gene: 362700 Rat
- Omim: 604277 Human
- SwissProt: Q9UBP0 Human
- SwissProt: B2RYN7 Rat
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Alternative names
- ADPSP antibody
- FSP 2 antibody
- FSP2 antibody
see all
Images
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All lanes : Anti-Spastin antibody [Sp 3G11/1] (ab31850) at 1/500 dilution
Lane 1 : Wild-type HEK-293T cell lysate
Lane 2 : SPAST knockout HEK293T cell lysate
Lane 3 : Wild-type HAP1 cell lysate
Lane 4 : SPAST knockout HAP1 cell lysate
Lysates/proteins at 20 µg per lane.
Secondary
All lanes : goat anti-rabbit HRP at 0.2 µg/ml
Performed under reducing conditions.ab31850 was shown to react with SPAST in wild-type HEK293T cells in Western blot with loss of signal observed in SPAST knockout cell line ab267238 (SPAST knockout cell lysate ab258698). Wild-type HEK293T and SPAST knockout cell lysates were subjected to SDS-PAGE. Membranes were blocked in 5% milk in TBST for 1 hr before incubation with ab31850 overnight at 4 °C at a 1/500 dilution. Blots were incubated with goat anti-rabbit HRP secondary antibodies at 0.2μg/mL before imaging.
These data were provided by YCharOS Inc., an open science company with the mission of characterizing commercially available antibody reagents for all human proteins. Abcam and YCharOS are working together to help address the reproducibility crisis by enabling the life science community to better evaluate commercially available antibodies.
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Immunoprecipitation of SPAST in HAP1 cells. Lysates were prepared and immunoprecipitation was performed using 1µg of ab31850 pre-coupled to Protein A beads. Samples were washed and processed for western blot with ab77144 at 1/500.
This data was kindly provided by the YCharOS Inc., an open science company with the mission of characterizing every commercially available antibody reagent. Abcam are working with YCharOS to support their mission of antibody characterisation using knock out cell lines.
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All lanes : Anti-Spastin antibody [Sp 3G11/1] (ab31850) at 1/500 dilution
Lane 1 : Wild-type HEK-293T cell lysate
Lane 2 : Spastin knockout HEK-293T cell lysate
Lane 3 : SH-SY5Y cell lysate
Lane 4 : HeLa cell lysate
Lysates/proteins at 20 µg per lane.
Performed under reducing conditions.
Observed band size: 55 kDa why is the actual band size different from the predicted?False colour image of Western blot: Anti-Spastin antibody [Sp 3G11/1] staining at 1/500 dilution, shown in green; Rabbit Anti-GAPDH antibody [EPR16891] (ab181602) loading control staining at 1/20000 dilution, shown in red. In Western blot, ab31850 was shown to bind specifically to Spastin. A band was observed at 55 kDa in wild-type HEK-293T cell lysates with no signal observed at this size in SPAST knockout cell line ab267238 (knockout cell lysate ab258698). To generate this image, wild-type and SPAST knockout HEK-293T 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-Mouse IgG H&L (IRDye® 800CW) preabsorbed (ab216772) and Goat anti-Rabbit IgG H&L (IRDye® 680RD) preabsorbed (ab216777) at 1/20000 dilution.
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Anti-Spastin antibody [Sp 3G11/1] (ab31850) at 1/500 dilution + rat brain synaptosome
Observed band size: 52 kDa why is the actual band size different from the predicted?
There are two splice isoforms of spastin (one without exon4) and two alternative ATG start sites, which may determine the localisation of the translate protein. 52kDa is the major band. -
IHC image of Spastin staining in Human lung adenocarcinoma formalin fixed paraffin embedded tissue section, performed on a Leica BondTM system using the standard protocol F. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH6, epitope retrieval solution 1) for 20 mins. The section was then incubated with ab31850, 10µg/ml, for 15 mins at room temperature and detected using an HRP conjugated compact polymer system. DAB was used as the chromogen. The section was then counterstained with haematoxylin and mounted with DPX.
For other IHC staining systems (automated and non-automated) customers should optimize variable parameters such as antigen retrieval conditions, primary antibody concentration and antibody incubation times.
Protocols
Datasheets and documents
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SDS download
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Datasheet download
References (3)
ab31850 has been referenced in 3 publications.
- Nicholson JM et al. Chromosome mis-segregation and cytokinesis failure in trisomic human cells. Elife 4:N/A (2015). PubMed: 25942454
- Daftary GS et al. A Novel Role for the AAA ATPase Spastin as a HOXA10 Transcriptional Corepressor in Ishikawa Endometrial Cells. Mol Endocrinol 25:1539-49 (2011). WB, ICC/IF, IHC-P, ChIP ; Human . PubMed: 21757506
- Salinas S et al. Human spastin has multiple microtubule-related functions. J Neurochem 95:1411-20 (2005). PubMed: 16219033