Anti-Spastin antibody [Sp 6C6] (ab77144)
Key features and details
- Mouse monoclonal [Sp 6C6] to Spastin
- Suitable for: IP, WB
- Knockout validated
- Reacts with: Rat, Human
- Isotype: IgG2a
Overview
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Product name
Anti-Spastin antibody [Sp 6C6]
See all Spastin primary antibodies -
Description
Mouse monoclonal [Sp 6C6] to Spastin -
Host species
Mouse -
Tested applications
Suitable for: IP, WBmore details -
Species reactivity
Reacts with: Rat, Human -
Immunogen
Recombinant full length protein (Human)
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Positive control
- WB: HEK293T cell lysate; Rat brain lysate. 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. Upon delivery aliquot and store at -20°C or -80°C. Avoid repeated freeze / thaw cycles. -
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 6C6 -
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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KO cell lines
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KO cell lysates
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Recombinant Protein
Applications
The Abpromise guarantee
Our Abpromise guarantee covers the use of ab77144 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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IP |
Use at an assay dependent concentration.
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WB | (1) |
1/500. Predicted molecular weight: 68 kDa.
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Notes |
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IP
Use at an assay dependent concentration. |
WB
1/500. Predicted molecular weight: 68 kDa. |
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 6C6] (ab77144) 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.
Predicted band size: 68 kDaab77144 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 ab77144 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 1µ of ab77144 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 6C6] (ab77144) at 1/500 dilution
Lane 1 : Wild-type HEK-293T cell lysate
Lane 2 : SPAST knockout HEK293T cell lysate
Lane 3 : Rat brain tissue lysate
Lysates/proteins at 20 µg per lane.
Secondary
All lanes : Goat Anti-Rabbit IgG H&L (IRDye® 680RD) preadsorbed (ab216777) at 1/10000 dilution
Predicted band size: 68 kDa
Observed band size: 52 kDa why is the actual band size different from the predicted?Lanes 1-3: Merged signal (red and green). Green - ab77144 observed at 52 kDa. Red - loading control ab181602 observed at 36 kDa.
ab77144 Anti-Spastin antibody [Sp 6C6] was shown to specifically react with Spastin in wild-type HEK293T cells. Loss of signal was observed when knockout cell line ab267238 (knockout cell lysate ab258698) was used. Wild-type and Spastin knockout samples were subjected to SDS-PAGE. ab77144 and Anti-GAPDH antibody[EPR16891] - Loading Control (ab181602) were incubated overnight at 4°C at 1 in 500 dilution and 1 in 20000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye® 680RD) preadsorbed (ab216777) and Goat anti-Mouse IgG H&L (IRDye® 800CW) preadsorbed (ab216772) secondary antibodies at 1 in 20000 dilution for 1 hour at room temperature before imaging.
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All lanes : Anti-Spastin antibody [Sp 6C6] (ab77144) at 1/1500 dilution
All lanes : Human induced pluripotent stem cell derived neuron whole cell lysate
Lysates/proteins at 20 µg per lane.
Secondary
All lanes : HRP-conjugated goat anti-mouse IgG polyclonal at 1/10 dilution
Developed using the ECL technique.
Performed under reducing conditions.
Predicted band size: 68 kDa
Observed band size: 54,58,62,68 kDa why is the actual band size different from the predicted?
Exposure time: 12 minutes9% SDS-PAGE.
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
References (5)
ab77144 has been referenced in 5 publications.
- Lombino FL et al. The Microtubule Severing Protein Katanin Regulates Proliferation of Neuronal Progenitors in Embryonic and Adult Neurogenesis. Sci Rep 9:15940 (2019). PubMed: 31685876
- Qiang L et al. Hereditary spastic paraplegia: gain-of-function mechanisms revealed by new transgenic mouse. Hum Mol Genet 28:1136-1152 (2019). PubMed: 30520996
- Havlicek S et al. Gene dosage-dependent rescue of HSP neurite defects in SPG4 patients' neurons. Hum Mol Genet 23:2527-41 (2014). WB ; Human . PubMed: 24381312
- Abrahamsen G et al. A patient-derived stem cell model of hereditary spastic paraplegia with SPAST mutations. Dis Model Mech 6:489-502 (2013). WB ; Human . PubMed: 23264559
- Salinas S et al. Human spastin has multiple microtubule-related functions. J Neurochem 95:1411-20 (2005). PubMed: 16219033