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  1. Link

    alas1-antibody-epr10247-mitochondrial-marker-ab154860.pdf

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Validated using a knockout cell lineRecombinantRabMAb

Recombinant Anti-Alas1 antibody [EPR10247] - Mitochondrial Marker (ab154860)

  • Datasheet
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Western blot - Anti-Alas1 antibody [EPR10247] - Mitochondrial Marker (ab154860)
  • Immunocytochemistry/ Immunofluorescence - Anti-Alas1 antibody [EPR10247] - Mitochondrial Marker (ab154860)
  • Western blot - Anti-Alas1 antibody [EPR10247] - Mitochondrial Marker (ab154860)
  • Western blot - Anti-Alas1 antibody [EPR10247] - Mitochondrial Marker (ab154860)
  • Western blot - Anti-Alas1 antibody [EPR10247] - Mitochondrial Marker (ab154860)
  • Immunocytochemistry/ Immunofluorescence - Anti-Alas1 antibody [EPR10247] - Mitochondrial Marker (ab154860)
  • Anti-Alas1 antibody [EPR10247] - Mitochondrial Marker (ab154860)

Key features and details

  • Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
  • Rabbit monoclonal [EPR10247] to Alas1 - Mitochondrial Marker
  • Suitable for: WB, ICC/IF
  • Knockout validated
  • Reacts with: Human

Conjugates logo Related conjugates and formulations

Carrier Free

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Overview

  • Product name

    Anti-Alas1 antibody [EPR10247] - Mitochondrial Marker
    See all Alas1 primary antibodies
  • Description

    Rabbit monoclonal [EPR10247] to Alas1 - Mitochondrial Marker
  • Host species

    Rabbit
  • Tested applications

    Suitable for: WB, ICC/IFmore details
    Unsuitable for: IHC-P
  • Species reactivity

    Reacts with: Human
  • Immunogen

    Synthetic peptide. This information is proprietary to Abcam and/or its suppliers.

  • Positive control

    • WB: HeLa, HEK-293, HepG2, K562, JAR and Raji cell lysate. ICC/IF: HepG2 cells.
  • 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
    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

  • Form

    Liquid
  • Storage instructions

    Shipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C. Avoid freeze / thaw cycle.
  • Storage buffer

    Preservative: 0.01% Sodium azide
    Constituents: 40% Glycerol, 0.05% BSA, 59% PBS
  • Concentration information loading...
  • Purity

    Protein A purified
  • Clonality

    Monoclonal
  • Clone number

    EPR10247
  • Isotype

    IgG
  • Research areas

    • Cardiovascular
    • Blood
    • Other
    • Signal Transduction
    • Metabolism
    • Mitochondrial
    • Metabolism
    • Pathways and Processes
    • Mitochondrial Metabolism
    • Mitochondrial markers

Associated products

  • Alternative Versions

    • Anti-Alas1 antibody [EPR10247] - BSA and Azide free (ab211925)
  • Compatible Secondaries

    • Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077)
    • Goat Anti-Rabbit IgG H&L (HRP) (ab205718)
    • Goat Anti-Rabbit IgG H&L (HRP) (ab97051)
  • Isotype control

    • Rabbit IgG, monoclonal [EPR25A] - Isotype Control (ab172730)
  • KO cell lines

    • Human ALAS1 knockout HEK-293T cell line (ab266473)
  • KO cell lysates

    • Human ALAS1 knockout HEK-293T cell lysate (ab257348)
  • Positive Controls

    • Hep G2 whole cell lysate (ab166833)
    • Raji whole cell lysate (ab30124)
    • K-562 whole cell lysate (ab7911)
  • Recombinant Protein

    • Recombinant Human Alas1 protein (ab114537)
  • Related Products

    • Mitochondrial Staining Kit - Red Fluorescence - Cytopainter (ab112145)
    • Mitochondrial Staining Kit - NIR Fluorescence - Cytopainter (ab176747)

Applications

The Abpromise guarantee

Our Abpromise guarantee covers the use of ab154860 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)
1/1000 - 1/10000. Predicted molecular weight: 71 kDa.
ICC/IF
1/100 - 1/500.
Notes
WB
1/1000 - 1/10000. Predicted molecular weight: 71 kDa.
ICC/IF
1/100 - 1/500.
Application notes
Is unsuitable for IHC-P.

Target

  • Pathway

    Porphyrin metabolism; protoporphyrin-IX biosynthesis; 5-aminolevulinate from glycine: step 1/1.
  • Sequence similarities

    Belongs to the class-II pyridoxal-phosphate-dependent aminotransferase family.
  • Cellular localization

    Mitochondrion matrix.
  • Target information above from: UniProt accession P13196 The UniProt Consortium
    The Universal Protein Resource (UniProt) in 2010
    Nucleic Acids Res. 38:D142-D148 (2010) .

    Information by UniProt
  • Database links

    • Entrez Gene: 211 Human
    • Omim: 125290 Human
    • SwissProt: P13196 Human
    • Unigene: 476308 Human
    • Alternative names

      • 5 aminolevulinate synthase antibody
      • 5 aminolevulinate synthase nonspecific mitochondrial antibody
      • 5 aminolevulinic acid synthase antibody
      • 5-aminolevulinate synthase antibody
      • 5-aminolevulinic acid synthase 1 antibody
      • Alas 1 antibody
      • ALAS 3 antibody
      • ALAS antibody
      • ALAS H antibody
      • ALAS HOUSEKEEPING TYPE antibody
      • ALAS N antibody
      • ALAS-H antibody
      • alaS1 antibody
      • ALAS3 antibody
      • ALASH antibody
      • Aminolevulinate delta synthase 1 antibody
      • Aminolevulinic acid synthase 1 antibody
      • Delta ALA synthetase antibody
      • Delta aminolevulinate synthase antibody
      • Delta-ALA synthase 1 antibody
      • Delta-aminolevulinate synthase 1 antibody
      • HEM1_HUMAN antibody
      • MIG 4 antibody
      • MIG4 antibody
      • Migration inducing protein 4 antibody
      • mitochondrial antibody
      • nonspecific antibody
      see all

    Images

    • Western blot - Anti-Alas1 antibody [EPR10247] - Mitochondrial Marker (ab154860)
      Western blot - Anti-Alas1 antibody [EPR10247] - Mitochondrial Marker (ab154860)
      All lanes : Anti-Alas1 antibody [EPR10247] - Mitochondrial Marker (ab154860) at 1/1000 dilution

      Lane 1 : Wild-type HeLa cell lysate
      Lane 2 : ALAS1 knockout HeLa cell lysate
      Lane 3 : HepG2 cell lysate
      Lane 4 : HEK-293 cell lysate

      Lysates/proteins at 20 µg per lane.

      Secondary
      All lanes : Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed (ab216773) at 1/10000 dilution

      Predicted band size: 71 kDa
      Observed band size: 71 kDa



      Lanes 1-4: Merged signal (red and green). Green - ab154860 observed at 71 kDa. Red - loading control ab8245 observed at 36 kDa. 

       ab154860 Anti-Alas1 antibody [EPR10247]  was shown to specifically react with Alas1 in wild-type HeLa cells. Loss of signal was observed when knockout cell line ab266473 (knockout cell lysate ab257348) was used.  Wild-type and Alas1 knockout samples were subjected to SDS-PAGE.  ab154860 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 20000 dilution for 1 hour at room temperature before imaging.

    • Immunocytochemistry/ Immunofluorescence - Anti-Alas1 antibody [EPR10247] - Mitochondrial Marker (ab154860)
      Immunocytochemistry/ Immunofluorescence - Anti-Alas1 antibody [EPR10247] - Mitochondrial Marker (ab154860)

      Immunocytochemistry/Immunofluorescence analysis of HepG2 cells labelling Alas1 with purified ab154860 at a dilution of 1/100. Cells were fixed with 4% paraformaldehyde and permeabilized with 0.1% Triton X-100. ab150077, an Alexa Fluor® 488-conjugated goat anti-rabbit IgG (1/1000) was used as the secondary antibody. DAPI (blue) was used as the nuclear counterstain. ab7291, a mouse anti-tubulin (1/1000) and ab150120, an Alexa Fluor® 594-conjugated goat anti-mouse IgG (1/1000) were also used.

      Control 1: primary antibody (1/100) and secondary antibody, ab150120, an Alexa Fluor® 594-conjugated goat anti-mouse IgG (1/1000).

      Control 2: ab7291 (1/1000) and secondary antibody, ab150077, an Alexa Fluor® 488-conjugated goat anti-rabbit IgG (1/1000).

    • Western blot - Anti-Alas1 antibody [EPR10247] - Mitochondrial Marker (ab154860)
      Western blot - Anti-Alas1 antibody [EPR10247] - Mitochondrial Marker (ab154860)
      Anti-Alas1 antibody [EPR10247] - Mitochondrial Marker (ab154860) at 1/10000 dilution (purified) + HepG2 whole cell lysate at 10 µg

      Secondary
      Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/100000 dilution

      Predicted band size: 71 kDa
      Observed band size: 71 kDa



      Blocking and dilution buffer: 5% NFDM/TBST
    • Western blot - Anti-Alas1 antibody [EPR10247] - Mitochondrial Marker (ab154860)
      Western blot - Anti-Alas1 antibody [EPR10247] - Mitochondrial Marker (ab154860)
      Anti-Alas1 antibody [EPR10247] - Mitochondrial Marker (ab154860) at 1/10000 dilution (purified) + Raji whole cell lysate at 10 µg

      Secondary
      Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/100000 dilution

      Predicted band size: 71 kDa
      Observed band size: 71 kDa



      Blocking and dilution buffer: 5% NFDM/TBST
    • Western blot - Anti-Alas1 antibody [EPR10247] - Mitochondrial Marker (ab154860)
      Western blot - Anti-Alas1 antibody [EPR10247] - Mitochondrial Marker (ab154860)
      All lanes : Anti-Alas1 antibody [EPR10247] - Mitochondrial Marker (ab154860) at 1/1000 dilution (unpurified)

      Lane 1 : HepG2 cell lysate
      Lane 2 : K562 cell lysate
      Lane 3 : JAR cell lysate
      Lane 4 : Raji cell lysate

      Lysates/proteins at 10 µg per lane.

      Secondary
      All lanes : HRP-conjugated goat anti-rabbit IgG at 1/2000 dilution

      Predicted band size: 71 kDa

    • Immunocytochemistry/ Immunofluorescence - Anti-Alas1 antibody [EPR10247] - Mitochondrial Marker (ab154860)
      Immunocytochemistry/ Immunofluorescence - Anti-Alas1 antibody [EPR10247] - Mitochondrial Marker (ab154860)

      Immunocytochemistry/Immunofluorescence analysis of HepG2 cells labeling Alas1 with unpurified ab154860 at a dilution of 1/250.

    • Anti-Alas1 antibody [EPR10247] - Mitochondrial Marker (ab154860)
      Anti-Alas1 antibody [EPR10247] - Mitochondrial Marker (ab154860)

    Protocols

    • Western blot protocols
    • Immunocytochemistry & immunofluorescence protocols

    Click here to view the general protocols

    Datasheets and documents

    • Datasheet download

      Download

    References (4)

    Publishing research using ab154860? Please let us know so that we can cite the reference in this datasheet.

    ab154860 has been referenced in 4 publications.

    • Acoba MG  et al. The mitochondrial carrier SFXN1 is critical for complex III integrity and cellular metabolism. Cell Rep 34:108869 (2021). PubMed: 33730581
    • Chen CP  et al. Synergistic antitumor activity of artesunate and HDAC inhibitors through elevating heme synthesis via synergistic upregulation of ALAS1 expression. Acta Pharm Sin B 9:937-951 (2019). PubMed: 31649844
    • Nakano T  et al. Hepatitis C virus core protein triggers abnormal porphyrin metabolism in human hepatocellular carcinoma cells. PLoS One 13:e0198345 (2018). PubMed: 29856826
    • Kubota Y  et al. Novel Mechanisms for Heme-dependent Degradation of ALAS1 Protein as a Component of Negative Feedback Regulation of Heme Biosynthesis. J Biol Chem N/A:N/A (2016). PubMed: 27496948

    Customer reviews and Q&As

    Show All Reviews Q&A
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    Western blot abreview for Anti-Alas1 antibody [EPR10247] - Mitochondrial Marker

    Excellent
    Abreviews
    Abreviews
    abreview image
    Application
    Western blot
    Sample
    Human Cell lysate - whole cell (HepG2 cells)
    Gel Running Conditions
    Reduced Denaturing (8% acrylamide gel)
    Loading amount
    10 µg
    Treatment
    with/without 0.1 mM succinylaceton
    Specification
    HepG2 cells
    Blocking step
    Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C
    Read More

    Abcam user community

    Verified customer

    Submitted Sep 21 2016

    Please note: All products are "FOR RESEARCH USE ONLY. NOT FOR USE IN DIAGNOSTIC PROCEDURES"
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