Anti-Spa-1 antibody [EPR14134] (ab189929)
![Anti-Spa-1 antibody [EPR14134] (ab189929)](https://www.abcam.com/ps/products/189/ab189929/Images/ab189929-389723-anti-spa-1-antibody-epr14134-western-blot-human-lysate.jpg "Anti-Spa-1 antibody [EPR14134] (ab189929)")
Key features and details
- Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
- Rabbit monoclonal [EPR14134] to Spa-1
- Suitable for: WB, IHC-P, ICC/IF, Flow Cyt
- Knockout validated
- Reacts with: Mouse, Human
Overview
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Product name
Anti-Spa-1 antibody [EPR14134]
See all Spa-1 primary antibodies -
Description
Rabbit monoclonal [EPR14134] to Spa-1 -
Host species
Rabbit -
Tested Applications & Species
See all applications and species dataApplication Species Flow Cyt HumanICC/IF HumanIHC-P HumanWB MouseHuman
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Immunogen
This information is proprietary to Abcam and/or its suppliers.
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Positive control
- WB: Human fetal thymus, HeLa, Raji, HEK-293T and Raw 264.7 lysates. IHC-P: Human kidney tissue. ICC/IF: A431 cells. Flow Cyt: 293T cells.
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General notes
Previously labelled as SIPA1.
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
EPR14134 -
Isotype
IgG -
Research areas
Images
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Western blot - Anti-Spa-1 antibody [EPR14134] (ab189929)All lanes : Anti-Spa-1 antibody [EPR14134] (ab189929) at 1/5000 dilution
Lane 1 : Wild-type HeLa cell lysate
Lane 2 : Spa-1 knockout HeLa cell lysate
Lane 3 : Raji cell lysate
Lane 4 : HEK-293 cell lysate
Lysates/proteins at 20 µg per lane.
Performed under reducing conditions.
Predicted band size: 112 kDaLanes 1-4: Merged signal (red and green). Green - ab189929 observed at 130 kDa. Red - loading control ab7291 observed at 50 kDa.
ab189929 Anti-Spa-1 antibody [EPR14134] was shown to specifically react with Spa-1 in wild-type HeLa cells. Loss of signal was observed when knockout cell line ab265434 (knockout cell lysate ab258189) was used. Wild-type and Spa-1 knockout samples were subjected to SDS-PAGE. ab189929 and Anti-alpha Tubulin antibody [DM1A] - Loading Control (ab7291) were incubated overnight at 4°C at 1 in 5000 dilution and 1 in 20000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye® 680RD) preadsorbed (ab216776) secondary antibodies at 1 in 20000 dilution for 1 hour at room temperature before imaging.
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![Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Spa-1 antibody [EPR14134] (ab189929)](https://www.abcam.com/ps/products/189/ab189929/Images/ab189929-226399-ip.png)
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Spa-1 antibody [EPR14134] (ab189929)Immunohistochemical analysis of formalin fixed, paraffin embedded Human kidney tissue labeling Spa-1 with ab189929 at 1/50 dilution, and Prediluted HRP Polymer for Rabbit IgG as a secondary antibody. Counterstained with Hematoxylin.
Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
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![Immunocytochemistry/ Immunofluorescence - Anti-Spa-1 antibody [EPR14134] (ab189929)](https://www.abcam.com/ps/products/189/ab189929/Images/ab189929-226398-ip.png)
Immunocytochemistry/ Immunofluorescence - Anti-Spa-1 antibody [EPR14134] (ab189929)Immunocytochemical analysis of A431 cells fixed in 4% paraformaldehyde labeling Spa-1 with ab189929 at 1/250 dilution, and Goat anti rabbit IgG(Alexa Fluor® 555) at 1/200 dilution as the secondary antibody. Counterstained with DAPI.
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![Western blot - Anti-Spa-1 antibody [EPR14134] (ab189929)](https://www.abcam.com/ps/products/189/ab189929/Images/ab189929-226401-wb.png)
Western blot - Anti-Spa-1 antibody [EPR14134] (ab189929)All lanes : Anti-Spa-1 antibody [EPR14134] (ab189929) at 1/5000 dilution
Lane 1 : Human fetal thymus
Lane 2 : Raji cell lysate
Lane 3 : 293T cell lysate
Lysates/proteins at 20 µg per lane.
Secondary
All lanes : Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated at 1/1000 dilution
Predicted band size: 112 kDa
Additional bands at: 130 kDa. We are unsure as to the identity of these extra bands.
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![Western blot - Anti-Spa-1 antibody [EPR14134] (ab189929)](https://www.abcam.com/ps/products/189/ab189929/Images/ab189929-226400-wb.png)
Western blot - Anti-Spa-1 antibody [EPR14134] (ab189929)Anti-Spa-1 antibody [EPR14134] (ab189929) at 1/1000 dilution + Raw264.7 at 10 µg
Secondary
Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated at 1/1000 dilution
Predicted band size: 112 kDa
Additional bands at: 130 kDa. We are unsure as to the identity of these extra bands.
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![Flow Cytometry - Anti-Spa-1 antibody [EPR14134] (ab189929)](https://www.abcam.com/ps/products/189/ab189929/Images/ab189929-226397-flow.png)
Flow Cytometry - Anti-Spa-1 antibody [EPR14134] (ab189929)Flow cytometric analysis of 293T cells fixed in 2% paraformaldehyde labeling Spa-1 with ab189929 at 1/400 dilution, and Goat anti rabbit IgG (FITC) at 1/150 dilution as the secondary antibody.
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![Anti-Spa-1 antibody [EPR14134] (ab189929)](https://www.abcam.com/ps/products/189/ab189929/Images/ab189929-7-benefits-of-recombinant-antibodies.png)
Anti-Spa-1 antibody [EPR14134] (ab189929)
