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Validated using a knockout cell lineRecombinantRabMAb

Recombinant Anti-SLC25A12 antibody [EPR16294] (ab200201)

Reviews (1) Submit a question References (2)
Western blot - Anti-SLC25A12 antibody [EPR16294] (ab200201)
  • Western blot - Anti-SLC25A12 antibody [EPR16294] (ab200201)
  • Western blot - Anti-SLC25A12 antibody [EPR16294] (ab200201)
  • Western blot - Anti-SLC25A12 antibody [EPR16294] (ab200201)
  • Immunoprecipitation - Anti-SLC25A12 antibody [EPR16294] (ab200201)
  • Immunoprecipitation - Anti-SLC25A12 antibody [EPR16294] (ab200201)
  • Anti-SLC25A12 antibody [EPR16294] (ab200201)

Key features and details

  • Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
  • Rabbit monoclonal [EPR16294] to SLC25A12
  • Suitable for: IP, WB
  • Knockout validated
  • Reacts with: Mouse, Rat, Human

Conjugates logo Related conjugates and formulations

Carrier Free

Overview

  • Product name

    Anti-SLC25A12 antibody [EPR16294]
    See all SLC25A12 primary antibodies

  • Description

    Rabbit monoclonal [EPR16294] to SLC25A12

  • Host species

    Rabbit

  • Tested applications

    Suitable for: IP, WBmore details

  • Species reactivity

    Reacts with: Mouse, Rat, Human

  • Immunogen

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

  • Positive control

    • WB: Human fetal heart and skeletal muscle lysates; HeLa, A431 C6, RAW 264.7, PC-12 and NIH/3T3 whole cell lysates; Mouse and rat brain, heart and kidney lysates. IP: Jurkat whole cell lysate.

  • 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.

Properties

Applications

The Abpromise guarantee

Our Abpromise guarantee covers the use of ab200201 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
IP
1/40.
WB (1)
1/1000. Detects a band of approximately 75 kDa (predicted molecular weight: 75 kDa).
Notes
IP
1/40.
WB
1/1000. Detects a band of approximately 75 kDa (predicted molecular weight: 75 kDa).

Target

  • Function

    Calcium-dependent mitochondrial aspartate and glutamate carrier. May have a function in the urea cycle.

  • Tissue specificity

    High levels in heart and skeletal muscle, low in brain and very low in kidney.

  • Involvement in disease

    Defects in SLC25A12 are the cause of aspartate-glutamate carrier 1 deficiency (AGC1D) [MIM:612949]; also called global cerebral hypomyelination. This syndrome consists of a child severe psychomotor retardation, hypotonia and hypomyelination of the central nervous system.

  • Sequence similarities

    Belongs to the mitochondrial carrier family.
    Contains 4 EF-hand domains.
    Contains 3 Solcar repeats.

  • Cellular localization

    Mitochondrion inner membrane.
  • Information by UniProt

  • Database links

    see all

  • Alternative names

    • AGC1 antibody
    • Araceli hiperlarga antibody
    • ARALAR antibody
    • ARALAR1 antibody
    • Calcium binding mitochondrial carrier superfamily member antibody
    • Calcium binding mitochondrial carrier superfamily member Aralar1 antibody
    • Calcium-binding mitochondrial carrier protein Aralar1 antibody
    • CMC1_HUMAN antibody
    • Mitochondrial aspartate glutamate carrier 1 antibody
    • SLC25A12 antibody
    • Solute carrier family 25 (aspartate/glutamate carrier) member 12 antibody
    • Solute carrier family 25 (mitochondrial carrier Aralar) member 12 antibody
    • Solute carrier family 25 member 12 antibody
    • solute carrier family 25, member 12 antibody
    see all

Images

  • Western blot - Anti-SLC25A12 antibody [EPR16294] (ab200201)
    Western blot - Anti-SLC25A12 antibody [EPR16294] (ab200201)

    Lane 1: Wild-type HAP1 cell lysate (20 µg)
    Lane 2: SLC25A12 knockout HAP1 cell lysate (20 µg)
    Lane 3: Human skeletal muscle lysate (20 µg)
    Lane 4: A431 cell lysate (20 µg)
    Lanes 1 - 4: Merged signal (red and green). Green - ab200201 observed at 72 kDa. Red - loading control, ab8245, observed at 37 kDa.

    ab200201 was shown to recognize SLC25A12 when SLC25A12 knockout samples were used, along with additional cross-reactive bands. Wild-type and SLC25A12 knockout samples were subjected to SDS-PAGE. ab200201 and ab8245 (loading control to GAPDH) were diluted to 1/1000 and 1/10000 respectively and incubated overnight at 4°C. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed ab216773 andGoat anti-Mouse IgG H&L (IRDye® 680RD) preadsorbed ab216776 secondary antibodies at 1/10000 dilution for 1 hour at room temperature before imaging.

  • Western blot - Anti-SLC25A12 antibody [EPR16294] (ab200201)
    Western blot - Anti-SLC25A12 antibody [EPR16294] (ab200201)
    All lanes : Anti-SLC25A12 antibody [EPR16294] (ab200201) at 1/5000 dilution

    Lane 1 : Human fetal heart lysate
    Lane 2 : Human skeletal muscle lysate

    Lysates/proteins at 20 µg per lane.

    Secondary
    All lanes : Anti-Rabbit IgG (HRP), specific to the non-reduced form of IgG at 1/1000 dilution

    Predicted band size: 75 kDa
    Observed band size: 75 kDa


    Exposure time: 10 seconds


    Blocking/Dilution buffer: 5% NFDM/TBST.

  • Western blot - Anti-SLC25A12 antibody [EPR16294] (ab200201)
    Western blot - Anti-SLC25A12 antibody [EPR16294] (ab200201)
    All lanes : Anti-SLC25A12 antibody [EPR16294] (ab200201) at 1/1000 dilution

    Lane 1 : HeLa (Human epithelial cells from cervix adenocarcinoma) whole cell lysate
    Lane 2 : A431 (Human epidermoid carcinoma) whole cell lysate

    Lysates/proteins at 10 µg per lane.

    Secondary
    All lanes : Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated at 1/1000 dilution

    Predicted band size: 75 kDa
    Observed band size: 75 kDa


    Exposure time: 3 minutes


    Blocking/Dilution buffer: 5% NFDM/TBST.

  • Western blot - Anti-SLC25A12 antibody [EPR16294] (ab200201)
    Western blot - Anti-SLC25A12 antibody [EPR16294] (ab200201)
    All lanes : Anti-SLC25A12 antibody [EPR16294] (ab200201) at 1/1000 dilution

    Lane 1 : Mouse brain lysate
    Lane 2 : Mouse heart lysate
    Lane 3 : Mouse kidney lysate
    Lane 4 : Rat brain lysate
    Lane 5 : Rat heart lysate
    Lane 6 : Rat kidney lysate
    Lane 7 : C6 (Rat glial tumor cells) whole cell lysate
    Lane 8 : RAW 264.7 (Mouse macrophage cells transformed with Abelson murine leukemia virus) whole cell lysate
    Lane 9 : PC-12 (Rat adrenal gland pheochromocytoma) whole cell lysate
    Lane 10 : NIH/3T3 (Mouse embyro fibroblast cells) whole cell lysate

    Lysates/proteins at 10 µg per lane.

    Secondary
    All lanes : Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated at 1/1000 dilution

    Predicted band size: 75 kDa
    Observed band size: 75 kDa


    Exposure time: 15 seconds


    Blocking/Dilution buffer: 5% NFDM/TBST.

  • Immunoprecipitation - Anti-SLC25A12 antibody [EPR16294] (ab200201)
    Immunoprecipitation - Anti-SLC25A12 antibody [EPR16294] (ab200201)

    SLC25A12 was immunoprecipitated from 1mg of Jurkat (Human T cell leukemia cells from peripheral blood) whole cell lysate with ab200201 at 1/40 dilution.

    Western blot was performed from the immunoprecipitate using ab200201 at 1/1000 dilution.

    Anti-Rabbit IgG (HRP), specific to the non-reduced form of IgG, was used as secondary antibody at 1/1500 dilution.

    Lane 1: Jurkat whole cell lysate 10 µg (Input).

    Lane 2: ab200201 IP in Jurkat whole cell lysate.

    Lane 3: Rabbit monoclonal IgG (ab172730) instead of ab200201 in Jurkat whole cell lysate.
    Blocking and dilution buffer and concentration: 5% NFDM/TBST.
    Exposure time: 30 second.

  • Immunoprecipitation - Anti-SLC25A12 antibody [EPR16294] (ab200201)
    Immunoprecipitation - Anti-SLC25A12 antibody [EPR16294] (ab200201)

    SLC25A12 was immunoprecipitated from 1mg of Jurkat (Human T cell leukemia cells from peripheral blood) whole cell lysate with ab200201 at 1/40 dilution.

    Western blot was performed from the immunoprecipitate using ab200201 at 1/1000 dilution (Panel A) or ab192244 at 1/1000 dilution (Panel B).

    Anti-Rabbit IgG (HRP), specific to the non-reduced form of IgG, was used as secondary antibody at 1/1500 dilution.

    Lane 1: Jurkat whole cell lysate 10 µg (Input).

    Lane 2: ab200201 IP in Jurkat whole cell lysate.

    Lane 3: Rabbit monoclonal IgG (ab172730) instead of ab200201 in Jurkat whole cell lysate.
    Blocking and dilution buffer and concentration: 5% NFDM/TBST.

    Exposure time (Panel A and Panel B): 30 second.

  • Anti-SLC25A12 antibody [EPR16294] (ab200201)
    Anti-SLC25A12 antibody [EPR16294] (ab200201)

References (2)

Publishing research using ab200201? Please let us know so that we can cite the reference in this datasheet.

ab200201 has been referenced in 2 publications.

  • Yasukawa K  et al. The microRNAs miR-302b and miR-372 regulate mitochondrial metabolism via the SLC25A12 transporter, which controls MAVS-mediated antiviral innate immunity. J Biol Chem 295:444-457 (2020). PubMed: 31767682
  • Sen NE  et al. Generation of an Atxn2-CAG100 knock-in mouse reveals N-acetylaspartate production deficit due to early Nat8l dysregulation. Neurobiol Dis 132:104559 (2019). PubMed: 31376479

Customer reviews and Q&As

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Western blot abreview for Anti-SLC25A12 antibody [EPR16294]

 Excellent
Abreviews
abreview image
Application
Western blot
Sample
Human Tissue lysate - other (Mitochondria isolated from cortex, human frozen)
Gel Running Conditions
Reduced Denaturing (4-12% Bis-Tris gradient gel. Laemmli buffer with 10% β-Mercaptoethanol. Samples were denatured at 55C for 10 min. Immobilon-FL transfer membranes. Overnight transfer at +4C.)
Loading amount
15 µg
Specification
Mitochondria isolated from cortex, human frozen
Blocking step
SEA Block Blocking Buffer as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 100% · Temperature: 22°C
Read More
The reviewer received a reward from Abcam’s Loyalty Program in thanks for submitting this Abreview and for helping the scientific community make better-informed decisions.

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Verified customer

Submitted Aug 15 2019

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