Recombinant Anti-Thrombospondin 1 antibody [EPR22927-54] (ab267388)
Key features and details
- Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
- Rabbit monoclonal [EPR22927-54] to Thrombospondin 1
- Suitable for: Flow Cyt (Intra), WB, IHC-P, ICC/IF, IP
- Reacts with: Mouse, Rat, Human
Related conjugates and formulations
Overview
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Product name
Anti-Thrombospondin 1 antibody [EPR22927-54]
See all Thrombospondin 1 primary antibodies -
Description
Rabbit monoclonal [EPR22927-54] to Thrombospondin 1 -
Host species
Rabbit -
Tested applications
Suitable for: Flow Cyt (Intra), WB, IHC-P, ICC/IF, IPmore details -
Species reactivity
Reacts with: Mouse, Rat, Human -
Immunogen
Recombinant fragment. This information is proprietary to Abcam and/or its suppliers.
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Positive control
- WB: 3T3-L1 starved with 0.4% serum for 24 hours, then cultivated with 15% serum for 6 hours, whole cell lysate. HUVEC and mouse platelet lysates and rat platelet whole cell lysate. IHC-P: Human spleen, human bone marrow and mouse spleen tissues. ICC/IF: HUVEC cells and PC-12 cells. Flow Cyt (intra): HUVEC, 3T3-L1 and PC-12 cells. IP: HUVEC and mouse platelets lysate.
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General notes
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: PBS, 0.05% BSA, 40% Glycerol (glycerin, glycerine) -
Concentration information loading...
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Purity
Protein A purified -
Clonality
Monoclonal -
Clone number
EPR22927-54 -
Isotype
IgG -
Research areas
Associated products
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Alternative Versions
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Compatible Secondaries
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Isotype control
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Related Products
Applications
The Abpromise guarantee
Our Abpromise guarantee covers the use of ab267388 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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Flow Cyt (Intra) |
1/50 - 1/500.
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WB |
1/1000. Predicted molecular weight: 129 kDa.
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IHC-P |
1/5000. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
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ICC/IF |
1/100.
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IP |
1/30.
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Notes |
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Flow Cyt (Intra)
1/50 - 1/500. |
WB
1/1000. Predicted molecular weight: 129 kDa. |
IHC-P
1/5000. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol. |
ICC/IF
1/100. |
IP
1/30. |
Target
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Function
Adhesive glycoprotein that mediates cell-to-cell and cell-to-matrix interactions. Binds heparin. May play a role in dentinogenesis and/or maintenance of dentin and dental pulp (By similarity). Ligand for CD36 mediating antiangiogenic properties. Plays a role in ER stress response, via its interaction with the activating transcription factor 6 alpha (ATF6) which produces adaptive ER stress response factors. -
Sequence similarities
Belongs to the thrombospondin family.
Contains 2 EGF-like domains.
Contains 1 laminin G-like domain.
Contains 1 TSP C-terminal (TSPC) domain.
Contains 3 TSP type-1 domains.
Contains 8 TSP type-3 repeats.
Contains 1 VWFC domain. -
Cellular localization
Endoplasmic reticulum. Sarcoplasmic reticulum. - Information by UniProt
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Database links
- Entrez Gene: 7057 Human
- Entrez Gene: 21825 Mouse
- Entrez Gene: 445442 Rat
- Omim: 188060 Human
- SwissProt: P07996 Human
- SwissProt: P35441 Mouse
- SwissProt: Q8CGB2 Mouse
- Unigene: 164226 Human
see all -
Alternative names
- Thbs1 antibody
- Thrombospondin-1 antibody
- TSP antibody
see all
Images
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Anti-Thrombospondin 1 antibody [EPR22927-54] (ab267388) at 1/1000 dilution + Rat platelet whole cell lysate at 20 µg
Secondary
Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/100000 dilution
Predicted band size: 129 kDa
Observed band size: 180,140 kDa why is the actual band size different from the predicted?
Exposure time: 0 secondBlocking buffer and concentration : 5% NFDM/TBST
Diluting buffer and concentration : 5% NFDM/TBSTThe full-length TSP 1 (180 kDa) and a ~140 kDa band, likely to be a TSP 1 isoform or fragment, are observed.
The molecular weight observed is consistent with what has been described in the literature (PMID:1426766, 27588705).
Exposure time : 5.5 seconds
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All lanes : Anti-Thrombospondin 1 antibody [EPR22927-54] (ab267388) at 1/1000 dilution
Lane 1 : HUVEC (human umbilical vein endothelial cell) whole cell lysate
Lane 2 : Mouse platelet lysate
Lysates/proteins at 20 µg per lane.
Secondary
All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/100000 dilution
Predicted band size: 129 kDa
Exposure time: 3 minutesThe full-length TSP 1 (180kDa) and a 160-kDa band, likely to be an TSP 1 isoform or fragment, are observed.
The molecular weight observed is consistent with what has been described in the literature (PMID:1426766, 27588705).
Blocking/Dilution buffer: 5% NFDM/TBST.
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All lanes : Anti-Thrombospondin 1 antibody [EPR22927-54] (ab267388) at 1/1000 dilution
Lane 1 : 3T3-L1 (mouse embryonic fibroblast) starved with 0.4% serum for 30 hours whole cell lysate
Lane 2 : 3T3-L1 starved with 0.4% serum for 24 hours, then cultivated with 15% serum for 6 hours. whole cell lysate
Lysates/proteins at 10 µg per lane.
Secondary
All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/100000 dilution
Predicted band size: 129 kDa
Exposure time: 3 minutesTSP 1 is a serum-responsive gene. Its express is elevated in quiescent 3T3 with serum addition.
The full-length TSP 1 (180kDa) and a 160-kDa band, likely to be an TSP 1 isoform or fragment, are observed.
The molecular weight observed is consistent with what has been described in the literature (PMID:1426766, 27588705).
Blocking/Dilution buffer: 5% NFDM/TBST.
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Thrombospondin 1 was immunoprecipitated from 0.35 mg HUVEC (human umbilical vein endothelial cell) whole cell lysate 10µg with ab267388 at 1/30 dilution (2µg in 0.35mg lysates). Western blot was performed on the immunoprecipitate using ab267388 at 1/1000 dilution. VeriBlot for IP Detection Reagent (HRP) (ab131366) was used at 1/5000 dilution.
Lane 1: HUVEC whole cell lysate 10µg.
Lane 2: ab267388 IP in HUVEC whole cell lysate.
Lane 3: Rabbit monoclonal IgG (ab172730) instead of ab267388 in HUVEC whole cell lysate.
Blocking and dilution buffer and concentration: 5% NFDM/TBST.
Exposure time: 8 seconds.
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Thrombospondin 1 was immunoprecipitated from 0.35 mg mouse platelets whole cell lysate 10µg with ab267388 at 1/30 dilution (2µg in 0.35mg lysates). Western blot was performed on the immunoprecipitate using ab267388 at 1/1000 dilution. VeriBlot for IP Detection Reagent (HRP) (ab131366) was used at 1/5000 dilution.
Lane 1: Mouse platelets whole cell lysate 10µg.
Lane 2: ab267388 IP in mouse platelets whole cell lysate.
Lane 3: Rabbit monoclonal IgG (ab172730) instead of ab267388 in mouse platelets whole cell lysate.
Blocking and dilution buffer and concentration: 5% NFDM/TBST.
Exposure time: 8 seconds.
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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Thrombospondin 1 antibody [EPR22927-54] (ab267388)
Immunohistochemical analysis of paraffin-embedded mouse spleen tissue labeling Thrombospondin 1 with ab267388 at 1/5000 dilution (0.1 µg/ml) followed by a ready to use Goat Anti-Rabbit IgG H&L (HRP Polymer). Positive staining on the megakaryocytes and platelets in the mouse spleen is observed. The section was incubated with ab267388 for 10 mins at room temperature. The immunostaining staining was performed on a Leica Biosystems BOND® RX instrument. Counterstained with hematoxylin.
Secondary antibody only control: Secondary antibody is a ready to use Goat Anti-Rabbit IgG H&L (HRP Polymer).
Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution 2) for 20 mins.
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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Thrombospondin 1 antibody [EPR22927-54] (ab267388)
Immunohistochemical analysis of paraffin-embedded human bone marrow tissue labeling Thrombospondin 1 with ab267388 at 1/5000 dilution (0.1 µg/ml) followed by a ready to use Goat Anti-Rabbit IgG H&L (HRP Polymer). Positive staining on the megakaryocytes in the human bone marrow (PMID: 28239144). The section was incubated with ab267388 for 10 mins at room temperature. The immunostaining staining was performed on a Leica Biosystems BOND® RX instrument. Counterstained with hematoxylin.
Secondary antibody only control: Secondary antibody is a ready to use Goat Anti-Rabbit IgG H&L (HRP Polymer).
Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution 2) for 20 mins.
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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Thrombospondin 1 antibody [EPR22927-54] (ab267388)
Immunohistochemical analysis of paraffin-embedded human spleen tissue labeling Thrombospondin 1 with ab267388 at 1/5000 dilution (0.1 µg/ml) followed by a ready to use Goat Anti-Rabbit IgG H&L (HRP Polymer). Positive staining on the platelets in the human spleen (PMID: 28239144). The section was incubated with ab267388 for 10 mins at room temperature. The immunostaining staining was performed on a Leica Biosystems BOND® RX instrument. Counterstained with hematoxylin.
Secondary antibody only control: Secondary antibody is a ready to use Goat Anti-Rabbit IgG H&L (HRP Polymer).
Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution 2) for 20 mins.
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Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized PC-12 (rat adrenal gland pheochromocytoma cell line) cells labelling Thrombospondin 1 with primary antibody anti-Thrombospondin 1 (ab267388) at 1/100 dilution, followed by Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed (ab150081) secondary antibody at 1/1000 dilution. Confocal image showing cytoplasmic staining in PC-12 cell line. Anti-alpha Tubulin antibody (DM1A) - Microtubule Marker (Alexa Fluor® 594) (ab195889) was used to counterstain tubulin at 1/200 dilution. The nuclear counter stain is DAPI (blue).
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Immunofluorescent analysis of 100% methanol-fixed, permeabilized HUVEC (human umbilical vein endothelial cell) cells labeling Thrombospondin 1 with ab267388 at 1/100 dilution (5 µg/ml), followed by ab150077 AlexaFluor®488 Goat anti-Rabbit secondary antibody at 1/1000 dilution (Green). Confocal image showing cytoplasmic staining in HUVEC cell line is observed. ab195889 Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor® 594) was used to counterstain tubulin at 1/200 dilution (Red). The nuclear counterstain was DAPI (Blue).
Secondary antibody only control: Secondary antibody is ab150077 AlexaFluor®488 Goat anti-Rabbit secondary at 1/1000 dilution.
100% methanol fixation is recommended.
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Intracellular Flow Cytometry analysis of PC-12 (Rat adrenal gland pheochromocytoma cell line) cells labeling Thrombospondin 1 with ab267388 at 1/500 dilution (Red). Cells were fixed with 4% Paraformaldehyde and permeabilised with 90% Methanol. A Goat Anti-rabbit IgG (Alexa Fluor® 488, ab150081) secondary antibody was used at 1/5000 dilution. Isotype control - Rabbit monoclonal IgG (Black) (ab172730). Unlabeled control - Cell without incubation with primary antibody and secondary antibody (Blue).
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Intracellular flow cytometric analysis of 4% paraformaldehyde fixed 90% methanol permeabilized 3T3-L1 (mouse embryonic fibroblast) starved with 0.4% serum for 24h, then cultured with 15% serum for 6h (Red) / Untreated control (Green) cells labeling Thrombospondin 1 with ab267388 at 1/50 (Red) compared with a Rabbit monoclonal IgG (ab172730) / Black isotype control and an unlabeled control (cells without incubation with primary antibody and secondary antibody) (Blue). A Goat anti rabbit IgG (Alexa Fluor® 488, ab150077) at 1/2000 dilution was used as the secondary antibody.
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Intracellular flow cytometric analysis of 4% paraformaldehyde fixed 90% methanol permeabilized HUVEC (human umbilical vein endothelial cell) cells labeling Thrombospondin 1 with ab267388 at 1/50 (Red) compared with a Rabbit monoclonal IgG (ab172730) / Black isotype control and an unlabeled control (cells without incubation with primary antibody and secondary antibody) (Blue). A Goat anti rabbit IgG (Alexa Fluor® 488, ab150077) at 1/2000 dilution was used as the secondary antibody.
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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Datasheet download
Certificate of Compliance
References (2)
ab267388 has been referenced in 2 publications.
- Chinnici CM et al. Extracellular Vesicle-Derived microRNAs of Human Wharton's Jelly Mesenchymal Stromal Cells May Activate Endogenous VEGF-A to Promote Angiogenesis. Int J Mol Sci 22:N/A (2021). PubMed: 33669517
- Chinnici CM et al. Small Extracellular Vesicles from Human Fetal Dermal Cells and Their MicroRNA Cargo: KEGG Signaling Pathways Associated with Angiogenesis and Wound Healing. Stem Cells Int 2020:8889379 (2020). PubMed: 32855639