Validated using a knockout cell line
Recombinant
RabMAb

Recombinant Anti-SPR antibody [EPR9290] (ab157194)

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Key features and details

  • Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
  • Rabbit monoclonal [EPR9290] to SPR
  • Suitable for: WB, IHC-P, ICC/IF, IP, Flow Cyt
  • Knockout validated
  • Reacts with: Human

Overview

  • Product name

    Anti-SPR antibody [EPR9290]
    See all SPR primary antibodies

  • Description

    Rabbit monoclonal [EPR9290] to SPR

  • Host species

    Rabbit

  • Tested applications

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

  • Species reactivity

    Reacts with: Human

  • Immunogen

    Synthetic peptide, corresponding to residues in Human SPR (UniProt: P35270).

  • Positive control

    • WB: A549, MCF7, HepG2 and HeLa (ab150035) cell lysates. IHC-P: Human thyroid gland carcinoma tissue. ICC/IF: HeLa cells. Flow Cyt: MCF7 cells.

  • General notes

    Mouse, Rat: We have preliminary internal testing data to indicate this antibody may not react with these species. Please contact us for more information.

     

    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.

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

    We are constantly working hard to ensure we provide our customers with best in class antibodies. As a result of this work we are pleased to now offer this antibody in purified format. We are in the process of updating our datasheets. The purified format is designated 'PUR' on our product labels. If you have any questions regarding this update, please contact our Scientific Support team.

Properties

Applications

Our Abpromise guarantee covers the use of ab157194 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 1/1000 - 1/10000. Predicted molecular weight: 28 kDa.
IHC-P 1/100 - 1/250. Perform heat mediated antigen retrieval before commencing with IHC staining protocol.
ICC/IF 1/100 - 1/250.
IP 1/10 - 1/100.
Flow Cyt 1/100 - 1/500.

ab172730 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody.

Target

  • Function

    Catalyzes the final one or two reductions in tetra-hydrobiopterin biosynthesis to form 5,6,7,8-tetrahydrobiopterin.

  • Involvement in disease

    Defects in SPR are the cause of dystonia DOPA-responsive due to sepiapterin reductase deficiency (DRDSPRD) [MIM:612716]. In the majority of cases, patients manifest progressive psychomotor retardation, dystonia and spasticity. Cognitive anomalies are also often present. The disease is due to severe dopamine and serotonin deficiencies in the central nervous system caused by a defect in BH4 synthesis. Dystonia is defined by the presence of sustained involuntary muscle contractions, often leading to abnormal postures.

  • Sequence similarities

    Belongs to the sepiapterin reductase family.

  • Post-translational
    modifications

    In vitro phosphorylation of Ser-213 by CaMK2 does not change kinetic parameters.

  • Cellular localization

    Cytoplasm.
  • Information by UniProt

  • Database links

    • Alternative names

      • OTTHUMP00000160199 antibody
      • SDR38C1 antibody
      • Sepiapterin reductase (7,8 dihydrobiopterin:NADP+ oxidoreductase) antibody
      • Sepiapterin reductase antibody
      • Short chain dehydrogenase/reductase family 38C, member 1 antibody
      • SPR antibody
      • SPRE_HUMAN antibody
      see all

    Images

    • Western blot - Anti-SPR antibody [EPR9290] (ab157194)
      Western blot - Anti-SPR antibody [EPR9290] (ab157194)
      All lanes : Anti-SPR antibody [EPR9290] (ab157194) at 1/1000 dilution

      Lane 1 : Wild-type HeLa cell lysate
      Lane 2 : SPR knockout HeLa cell lysate
      Lane 3 : MCF7 cell lysate

      Lysates/proteins at 20 µg per lane.

      Performed under reducing conditions.

      Predicted band size: 28 kDa
      Observed band size: 26 kDa
      why is the actual band size different from the predicted?



      Lanes 1-3: Merged signal (red and green). Green - ab157194 observed at 26 kDa. Red - loading control, ab8245 observed at 37 kDa.

      ab157194 Anti-SPR antibody [EPR9290] was shown to specifically react with SPR in wild-type HeLa cells. Loss of signal was observed when knockout cell line ab264681 (knockout cell lysate ab258700) was used. Wild-type and SPR knockout samples were subjected to SDS-PAGE. ab157194 and Anti-GAPDH antibody [6C5] - Loading Control (ab8245) were incubated overnight at 4°C at 1 in 1000 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 10000 dilution for 1 hour at room temperature before imaging.

       

    • Western blot - Anti-SPR antibody [EPR9290] (ab157194)
      Western blot - Anti-SPR antibody [EPR9290] (ab157194)
      All lanes : Anti-SPR antibody [EPR9290] (ab157194) at 1/1000 dilution

      Lane 1 : A549 cell lysate
      Lane 2 : MCF7 cell lysate
      Lane 3 : HepG2 cell lysate
      Lane 4 : HeLa cell lysate

      Lysates/proteins at 10 µg per lane.

      Secondary
      All lanes : Goat anti-rabbit HRP at 1/2000 dilution

      Predicted band size: 28 kDa

    • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-SPR antibody [EPR9290] (ab157194)
      Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-SPR antibody [EPR9290] (ab157194)

      ab157194 showing +ve staining in Human normal pancreas.

      Perform heat mediated antigen retrieval before commencing with IHC staining protocol.

    • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-SPR antibody [EPR9290] (ab157194)
      Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-SPR antibody [EPR9290] (ab157194)

      ab157194 showing +ve staining in Human heart.

      Perform heat mediated antigen retrieval before commencing with IHC staining protocol.

    • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-SPR antibody [EPR9290] (ab157194)
      Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-SPR antibody [EPR9290] (ab157194)

      ab157194 showing +ve staining in Human endometrial carcinoma.

      Perform heat mediated antigen retrieval before commencing with IHC staining protocol.

    • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-SPR antibody [EPR9290] (ab157194)
      Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-SPR antibody [EPR9290] (ab157194)

      ab157194 showing +ve staining in Human colonic adenocarcinoma.

      Perform heat mediated antigen retrieval before commencing with IHC staining protocol.

    • Flow Cytometry - Anti-SPR antibody [EPR9290] (ab157194)
      Flow Cytometry - Anti-SPR antibody [EPR9290] (ab157194)
      Flow cytometic analysis of permeabilized MCF7 cells labeling TPST2 with ab157194 at 1/100 (red) or a rabbit IgG (negative) (green).
    • Immunocytochemistry/ Immunofluorescence - Anti-SPR antibody [EPR9290] (ab157194)
      Immunocytochemistry/ Immunofluorescence - Anti-SPR antibody [EPR9290] (ab157194)
      Immunofluorescent analysis of HeLa cells labeling TPST2 with ab157194 at 1/100 dilution.
    • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-SPR antibody [EPR9290] (ab157194)
      Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-SPR antibody [EPR9290] (ab157194)

      Immunohistochemical analysis of paraffin-eEmbedded Human thyroid gland carcinoma tissue labeling TPST2 with ab157194 at 1/100 dilution.

      Perform heat mediated antigen retrieval before commencing with IHC staining protocol.

    • Immunoprecipitation - Anti-SPR antibody [EPR9290] (ab157194)
      Immunoprecipitation - Anti-SPR antibody [EPR9290] (ab157194)
      Detection of TPST2 by Western Blot of Immunprecipitate. MCF7 cell lysate immunoprecipitated using ab157194 at 1/10 dilution.

    References

    Publishing research using ab157194? Please let us know so that we can cite the reference in this datasheet.

    ab157194 has been referenced in 3 publications.

    • Wu Y  et al. Sepiapterin reductase promotes hepatocellular carcinoma progression via FoxO3a/Bim signaling in a nonenzymatic manner. Cell Death Dis 11:248 (2020). PubMed: 32312975
    • Mooney MR  et al. Anti-tumor effect of sulfasalazine in neuroblastoma. Biochem Pharmacol 162:237-249 (2019). PubMed: 30639262
    • Sallin O  et al. Semisynthetic biosensors for mapping cellular concentrations of nicotinamide adenine dinucleotides. Elife 7:N/A (2018). PubMed: 29809136

    Customer reviews and Q&As

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    Western blot abreview for Anti-SPR antibody [EPR9290]

     Good
    Abreviews
    abreview image
    Application
    Western blot
    Sample
    Rabbit Tissue lysate - whole (brain)
    Gel Running Conditions
    Non-reduced Denaturing
    Loading amount
    0.2 µg
    Treatment
    hypoxic 40 minutes at E25
    Specification
    brain
    Read More

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    Verified customer

    Submitted Jan 25 2018

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