Recombinant Anti-Thrombospondin 1 antibody [EPR22927-54] - BSA and Azide free (ab267397)
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 - BSA and Azide free
- Suitable for: Flow Cyt (Intra), WB, ICC/IF, IHC-P, IP
- Reacts with: Mouse, Rat, Human
Related conjugates and formulations
Overview
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Product name
Anti-Thrombospondin 1 antibody [EPR22927-54] - BSA and Azide free
See all Thrombospondin 1 primary antibodies -
Description
Rabbit monoclonal [EPR22927-54] to Thrombospondin 1 - BSA and Azide free -
Host species
Rabbit -
Tested applications
Suitable for: Flow Cyt (Intra), WB, ICC/IF, IHC-P, 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, human cervical carcinoma 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
ab267397 is the carrier-free version of ab267388.
Our carrier-free antibodies are typically supplied in a PBS-only formulation, purified and free of BSA, sodium azide and glycerol. The carrier-free buffer and high concentration allow for increased conjugation efficiency.
This conjugation-ready format is designed for use with fluorochromes, metal isotopes, oligonucleotides, and enzymes, which makes them ideal for antibody labelling, functional and cell-based assays, flow-based assays (e.g. mass cytometry) and Multiplex Imaging applications.
Use our conjugation kits for antibody conjugates that are ready-to-use in as little as 20 minutes with <1 minute hands-on-time and 100% antibody recovery: available for fluorescent dyes, HRP, biotin and gold.
This product is compatible with the Maxpar® Antibody Labeling Kit from Fluidigm, without the need for antibody preparation. Maxpar® is a trademark of Fluidigm Canada Inc.
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. Do Not Freeze. -
Storage buffer
pH: 7.2
Constituent: PBS -
Carrier free
Yes -
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 ab267397 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) |
Use at an assay dependent concentration.
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WB |
Use at an assay dependent concentration. Predicted molecular weight: 129 kDa.
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ICC/IF |
Use at an assay dependent concentration.
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IHC-P |
Use at an assay dependent concentration. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
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IP |
Use at an assay dependent concentration.
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Notes |
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Flow Cyt (Intra)
Use at an assay dependent concentration. |
WB
Use at an assay dependent concentration. Predicted molecular weight: 129 kDa. |
ICC/IF
Use at an assay dependent concentration. |
IHC-P
Use at an assay dependent concentration. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol. |
IP
Use at an assay dependent concentration. |
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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Immunofluorescent analysis of 100% Methanol-fixed, 0.1% TritonX-100 permeabilized PC-12 (rat adrenal gland pheochromocytoma cell) cells labelling Thrombospondin 1 with ab267388 at 1/100 (5.3 μg/ml) dilution, followed by ab150077 AlexaFluor® 488 Goat anti-Rabbit secondary antibody at 1/1000 (2 μg/ml) dilution (Green). Confocal image showing cytoplasmic staining in PC-12 cell line. Image was taken with a confocal microscope (Leica-Microsystems, TCS SP8). ab195889 Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor® 594) was used to counterstain tubulin at 1/200 (2.5 μg/ml) 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 (2 μg/ml) dilution.
This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol and sodium azide ab267388.
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Immunohistochemical analysis of paraffin-embedded Human cervical carcinoma tissue labelling Thrombospondin 1 with ab267388 at 1/5000 (0.101 μg/ml) followed by a Rabbit specific IHC polymer detection kit HRP/DAB (ab209101) at a Ready to use dilution. Positive staining on extracellular matrix of human cervical carcinoma. The section was incubated with ab267388 for 30 mins at room temperature. The immunostaining was performed on a Leica Biosystems BOND® RX instrument Counterstained with Hematoxylin.
Secondary antibody only control: Secondary antibody is Rabbit specific IHC polymer detection kit HRP/DAB (ab209101) at Ready to use dilution.
Heat mediated antigen retrieval was performed with Tris-EDTA buffer (pH 9.0, Epitope Retrieval Solution2) for 20 mins.
This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol and sodium azide ab267388.
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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?Blocking 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
This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol and sodium azide ab267388.
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This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol and sodium azide ab267388.
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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This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol and sodium azide ab267388.
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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This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol and sodium azide 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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This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol and sodium azide 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.
-
This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol and sodium azide 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.
-
This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol and sodium azide ab267388.
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).
This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol and sodium azide ab267388.
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This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol and sodium azide ab267388.
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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This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol and sodium azide ab267388.
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 (0)
ab267397 has not yet been referenced specifically in any publications.