Recombinant
RabMAb

Recombinant Anti-Thrombospondin 1 antibody [EPR22927-54] - BSA and Azide free (ab267397)

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Overview

  • 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: WB, ICC/IF, IHC-P, IP, Flow Cytmore details

  • Species reactivity

    Reacts with: Mouse, Human

  • Immunogen

    Recombinant fragment within Mouse Thrombospondin 1 aa 500 to the C-terminus. The exact sequence is proprietary.
    Database link: P35441

  • 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. IHC-P: Human spleen, human bone marrow and mouse spleen tissues. ICC/IF: HUVEC cells. FC: HUVEC and 3T3-L1 cells. IP: HUVEC and mouse platelets lysate.

  • General notes

    ab267397 is the carrier-free version of ab267388. This format is designed for use in antibody labeling, including fluorochromes, metal isotopes, oligonucleotides, enzymes.

    Our carrier-free formats are supplied in a buffer free of BSA, sodium azide and glycerol for higher conjugation efficiency.

    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.

    ab267397 is compatible with the Maxpar® Antibody Labeling Kit from Fluidigm.

    Maxpar® is a trademark of Fluidigm Canada Inc.??

    Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMAb® patents.

    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
    For more information see here.

Properties

Applications

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
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.
Flow Cyt Use at an assay dependent concentration.

Target

  • Relevance

    Thrombospondin is a regulator of many biological processes including cell growth, adhesion, migration, platelet aggregation, and fibrin deposition and lysis. It interacts with a number of plasma proteins such as fibrinogen and plasminogen and co-polymerizes with fibrin in clot formation. Thrombospondin also has multiple binding sites that interact with molecules such as fibronectin, collagens, laminin and heparan sulphate proteoglycans as well as binding growth factors such as TGF-beta1. Thrombospondin exerts an anti-adhesive effect which leads to cell rounding and detachment.

  • Database links

    • Alternative names

      • THBS1 antibody
      • TSP antibody
      • Tsp1 antibody

    Images

    • Immunoprecipitation - Anti-Thrombospondin 1 antibody [EPR22927-54] - BSA and Azide free (ab267397)
      Immunoprecipitation - Anti-Thrombospondin 1 antibody [EPR22927-54] - BSA and Azide free (ab267397)

      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.

       

    • Immunoprecipitation - Anti-Thrombospondin 1 antibody [EPR22927-54] - BSA and Azide free (ab267397)
      Immunoprecipitation - Anti-Thrombospondin 1 antibody [EPR22927-54] - BSA and Azide free (ab267397)

      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.

       

    • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Thrombospondin 1 antibody [EPR22927-54] - BSA and Azide free (ab267397)
      Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Thrombospondin 1 antibody [EPR22927-54] - BSA and Azide free (ab267397)

      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.

    • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Thrombospondin 1 antibody [EPR22927-54] - BSA and Azide free (ab267397)
      Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Thrombospondin 1 antibody [EPR22927-54] - BSA and Azide free (ab267397)

      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.

    • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Thrombospondin 1 antibody [EPR22927-54] - BSA and Azide free (ab267397)
      Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Thrombospondin 1 antibody [EPR22927-54] - BSA and Azide free (ab267397)

      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.

    • Immunocytochemistry/ Immunofluorescence - Anti-Thrombospondin 1 antibody [EPR22927-54] - BSA and Azide free (ab267397)
      Immunocytochemistry/ Immunofluorescence - Anti-Thrombospondin 1 antibody [EPR22927-54] - BSA and Azide free (ab267397)

      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.

    • Flow Cytometry - Anti-Thrombospondin 1 antibody [EPR22927-54] - BSA and Azide free (ab267397)
      Flow Cytometry - Anti-Thrombospondin 1 antibody [EPR22927-54] - BSA and Azide free (ab267397)

      This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol and sodium azide ab267388.

      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.

    • Flow Cytometry - Anti-Thrombospondin 1 antibody [EPR22927-54] - BSA and Azide free (ab267397)
      Flow Cytometry - Anti-Thrombospondin 1 antibody [EPR22927-54] - BSA and Azide free (ab267397)

      This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol and sodium azide ab267388.

      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.

    References

    ab267397 has not yet been referenced specifically in any publications.

    Publishing research using ab267397? Please let us know so that we can cite the reference in this datasheet.

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