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  1. Link

    products/primary-antibodies/mttfa-antibody-18g102b2e11-mitochondrial-marker-ab119684.pdf

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Tags & Cell Markers Subcellular Markers Organelles Mitochondria
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Anti-mtTFA antibody [18G102B2E11] - Mitochondrial Marker (ab119684)

  • Datasheet
Reviews (1)Q&A (3)References (21)

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Western blot - Anti-mtTFA antibody [18G102B2E11] - Mitochondrial Marker (ab119684)
  • Immunocytochemistry/ Immunofluorescence - Anti-mtTFA antibody [18G102B2E11] - Mitochondrial Marker (ab119684)
  • Flow Cytometry - Anti-mtTFA antibody [18G102B2E11] - Mitochondrial Marker (ab119684)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-mtTFA antibody [18G102B2E11] - Mitochondrial Marker (ab119684)

Key features and details

  • Mouse monoclonal [18G102B2E11] to mtTFA - Mitochondrial Marker
  • Suitable for: IHC-P, WB, Flow Cyt, ICC/IF
  • Reacts with: Human, Recombinant fragment
  • Isotype: IgG2b

Conjugates logo Related conjugates and formulations

Alexa Fluor® 488 Alexa Fluor® 647

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Protein
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Recombinant Human mtTFA protein (ab103784)
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Anti-Complex I subunit 8 kDa antibody [17C8E4E11] (ab110245)
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Alexa Fluor® 488 Anti-MTCO1 antibody [1D6E1A8] (ab154477)

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Overview

  • Product name

    Anti-mtTFA antibody [18G102B2E11] - Mitochondrial Marker
    See all mtTFA primary antibodies
  • Description

    Mouse monoclonal [18G102B2E11] to mtTFA - Mitochondrial Marker
  • Host species

    Mouse
  • Tested applications

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

    Reacts with: Human, Recombinant fragment
    Does not react with: Mouse, Rat
  • Immunogen

    Full length protein. This information is proprietary to Abcam and/or its suppliers.

  • Positive control

    • IHC-P: human liver FFPE tissue sections
  • General notes

    This antibody clone is manufactured by Abcam. If you require a custom buffer formulation or conjugation for your experiments, please contact orders@abcam.com.

    The Life Science industry has been in the grips of a reproducibility crisis for a number of years. Abcam is leading the way in addressing this with our range of recombinant monoclonal antibodies and knockout edited cell lines for gold-standard validation. Please check that this product meets your needs before purchasing.

    If you have any questions, special requirements or concerns, please send us an inquiry and/or contact our Support team ahead of purchase. Recommended alternatives for this product can be found below, along with publications, customer reviews and Q&As

    Product was previously marketed under the MitoSciences sub-brand.

Properties

  • Form

    Liquid
  • Storage instructions

    Shipped at 4°C. Store at +4°C. Do Not Freeze.
  • Storage buffer

    pH: 7.5
    Preservative: 0.02% Sodium azide
    Constituent: HEPES buffered saline
  • Concentration information loading...
  • Purity

    Ammonium Sulphate Precipitation
  • Purification notes

    Purity is near homogeneity as judged by SDS-PAGE. ab119684 was produced in vitro using hybridomas grown in serum-free medium, and then concentrated by ammonium sulfate precipitation.
  • Clonality

    Monoclonal
  • Clone number

    18G102B2E11
  • Isotype

    IgG2b
  • Light chain type

    kappa
  • Research areas

    • Tags & Cell Markers
    • Subcellular Markers
    • Organelles
    • Mitochondria
    • Signal Transduction
    • Metabolism
    • Mitochondrial
    • Epigenetics and Nuclear Signaling
    • Transcription
    • Transcription Factors
    • Cardiovascular
    • Heart
    • Cardiogenesis
    • Transcription factors/regulators
    • Metabolism
    • Pathways and Processes
    • Mitochondrial Metabolism
    • Mitochondrial Biogenesis
    • Metabolism
    • Pathways and Processes
    • Metabolic signaling pathways
    • Nucleotide metabolism
    • Molecular processes
    • Mitochondrial transcription
    • Metabolism
    • Pathways and Processes
    • Mitochondrial Metabolism
    • Mitochondrial markers
    • Metabolism
    • Types of disease
    • Cancer
    • Neuroscience
    • Processes

Associated products

  • Alternative Versions

    • Alexa Fluor® 647 Anti-mtTFA antibody [18G102B2E11] (ab198106)
    • Alexa Fluor® 488 Anti-mtTFA antibody [18G102B2E11] (ab198308)
  • Compatible Secondaries

    • Goat Anti-Mouse IgG H&L (Alexa Fluor® 488) (ab150113)
    • Goat Anti-Mouse IgG H&L (HRP) (ab205719)
  • Conjugation kits

    • PE / R-Phycoerythrin Conjugation Kit - Lightning-Link® (ab102918)
    • APC Conjugation Kit - Lightning-Link® (ab201807)
  • Isotype control

    • Mouse IgG2b, kappa monoclonal [7E10G10] - Isotype Control (ab170192)
  • Recombinant Protein

    • Recombinant Human mtTFA protein (ab103784)

Applications

The Abpromise guarantee

Our Abpromise guarantee covers the use of ab119684 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
IHC-P
Use a concentration of 5 µg/ml. Perform heat mediated antigen retrieval before commencing with IHC staining protocol.
WB (1)
Use a concentration of 1 µg/ml. Predicted molecular weight: 29 kDa.
Flow Cyt
Use a concentration of 1 µg/ml.

ab170192 - Mouse monoclonal IgG2b, is suitable for use as an isotype control with this antibody.

ICC/IF
Use a concentration of 1 µg/ml.
Notes
IHC-P
Use a concentration of 5 µg/ml. Perform heat mediated antigen retrieval before commencing with IHC staining protocol.
WB
Use a concentration of 1 µg/ml. Predicted molecular weight: 29 kDa.
Flow Cyt
Use a concentration of 1 µg/ml.

ab170192 - Mouse monoclonal IgG2b, is suitable for use as an isotype control with this antibody.

ICC/IF
Use a concentration of 1 µg/ml.

Target

  • Function

    Binds to the mitochondrial light strand promoter and functions in mitochondrial transcription regulation. Required for accurate and efficient promoter recognition by the mitochondrial RNA polymerase. Promotes transcription initiation from the HSP1 and the light strand promoter by binding immediately upstream of transcriptional start sites. Is able to unwind and bend DNA. Required for maintenance of normal levels of mitochondrial DNA. May play a role in organizing and compacting mitochondrial DNA. target DNA. Interacts with TFB1M and TFB2M.
  • Sequence similarities

    Contains 2 HMG box DNA-binding domains.
  • Cellular localization

    Mitochondrion.
  • Target information above from: UniProt accession Q00059 The UniProt Consortium
    The Universal Protein Resource (UniProt) in 2010
    Nucleic Acids Res. 38:D142-D148 (2010) .

    Information by UniProt
  • Database links

    • Entrez Gene: 7019 Human
    • Omim: 600438 Human
    • SwissProt: Q00059 Human
    • Unigene: 642966 Human
    • Alternative names

      • anscription factor 6-like 1 antibody
      • Mitochondrial transcription factor 1 antibody
      • mitochondrial transcription factor A antibody
      • MtTF1 antibody
      • mtTFA antibody
      • TCF 6 antibody
      • TCF-6 antibody
      • TCF6 antibody
      • TCF6L1 antibody
      • TCF6L2 antibody
      • TCF6L3 antibody
      • TFAM antibody
      • TFAM_HUMAN antibody
      • Transcription factor 6 antibody
      • Transcription factor 6 like 2 (mitochondrial transcription factor) antibody
      • Transcription factor 6 like 2 antibody
      • Transcription factor 6-like 2 antibody
      • transcription factor 6-like 3 antibody
      • Transcription factor A, mitochondrial antibody
      • Transcription factor A, mitochondrial antibody
      • Transcription factor A, mitochondrial precursor antibody
      see all

    Images

    • Western blot - Anti-mtTFA antibody [18G102B2E11] - Mitochondrial Marker (ab119684)
      Western blot - Anti-mtTFA antibody [18G102B2E11] - Mitochondrial Marker (ab119684)
      All lanes : Anti-mtTFA antibody [18G102B2E11] - Mitochondrial Marker (ab119684) at 1 µg/ml

      Lane 1 : recombinant mtTFA (ab103784) at 0.025 µg
      Lane 2 : HeLa whole cell lysate at 30 µg

      Secondary
      All lanes : HRP-conjugated Goat polyclonal to mouse IgG at 1/5000 dilution

      Predicted band size: 29 kDa

    • Immunocytochemistry/ Immunofluorescence - Anti-mtTFA antibody [18G102B2E11] - Mitochondrial Marker (ab119684)
      Immunocytochemistry/ Immunofluorescence - Anti-mtTFA antibody [18G102B2E11] - Mitochondrial Marker (ab119684)
      Immunocytochemistry image of ab119684 stained human HDFn cells. The cells were paraformaldehyde fixed (4%, 20 min) and Triton X-100 permeabilized (0.1%, 15min). The cells were incubated with the antibody (18G102B2E11, 1µg/ml) for 2h at room temperature. The secondary antibody was (green) 488 goat anti-mouse IgG (H+L) used at a 1/1000 dilution for 1h. 10% Goat serum was used as the blocking agent for all blocking steps. DAPI was used to stain the cell nuclei (blue). mtTFA localizes to the mitochondria.
    • Flow Cytometry - Anti-mtTFA antibody [18G102B2E11] - Mitochondrial Marker (ab119684)
      Flow Cytometry - Anti-mtTFA antibody [18G102B2E11] - Mitochondrial Marker (ab119684)
      HeLa cells were stained with 2 µg/mL mtTFA antibody (ab119684) (blue) or an equal amount of an isotype control antibody (red) and analyzed by flow cytometry.
    • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-mtTFA antibody [18G102B2E11] - Mitochondrial Marker (ab119684)
      Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-mtTFA antibody [18G102B2E11] - Mitochondrial Marker (ab119684)
      IHC image of mtTFA staining in human liver HCC formalin fixed paraffin embedded tissue section, performed on a Leica BondTM system using the standard protocol F. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH6, epitope retrieval solution 1) for 20 mins. The section was then incubated with ab119684, 5µg/ml, for 15 mins at room temperature and detected using an HRP conjugated compact polymer system. DAB was used as the chromogen. The section was then counterstained with haematoxylin and mounted with DPX.

      For other IHC staining systems (automated and non-automated) customers should optimize variable parameters such as antigen retrieval conditions, primary antibody concentration and antibody incubation times.

    Protocols

    • Flow cytometry protocols
    • Immunohistochemistry protocols
    • Immunocytochemistry & immunofluorescence protocols
    • Western blot protocols

    Click here to view the general protocols

    Datasheets and documents

    • Datasheet download

      Download

    References (21)

    Publishing research using ab119684? Please let us know so that we can cite the reference in this datasheet.

    ab119684 has been referenced in 21 publications.

    • Alexander JF  et al. Nasal administration of mitochondria reverses chemotherapy-induced cognitive deficits. Theranostics 11:3109-3130 (2021). PubMed: 33537077
    • Hao Z  et al. N6-Deoxyadenosine Methylation in Mammalian Mitochondrial DNA. Mol Cell 78:382-395.e8 (2020). PubMed: 32183942
    • Maekawa H  et al. Mitochondrial Damage Causes Inflammation via cGAS-STING Signaling in Acute Kidney Injury. Cell Rep 29:1261-1273.e6 (2019). PubMed: 31665638
    • Li H  et al. Mitochondrial dysfunction and mitophagy defect triggered by heterozygous GBA mutations. Autophagy 15:113-130 (2019). PubMed: 30160596
    • Li Q  et al. Ochratoxin A causes mitochondrial dysfunction, apoptotic and autophagic cell death and also induces mitochondrial biogenesis in human gastric epithelium cells. Arch Toxicol 93:1141-1155 (2019). PubMed: 30903243
    View all Publications for this product

    Customer reviews and Q&As

    Show All Reviews Q&A
    Submit a review Submit a question

    1-4 of 4 Abreviews or Q&A

    Western blot abreview for Anti-mtTFA antibody [18G102B2E11]

    Excellent
    Abreviews
    Abreviews
    abreview image
    Application
    Western blot
    Sample
    Human Tissue lysate - whole (Muscle)
    Gel Running Conditions
    Reduced Denaturing (12%)
    Loading amount
    15 µg
    Specification
    Muscle
    Blocking step
    Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°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.

    Abcam user community

    Verified customer

    Submitted Jul 10 2015

    Question

    The samples are prepared from schwann cells and DRG neurons and pancreatic cancer cell lines (one human, one rat origin, one mouse origin ) we chose the antibodies according to the reactivity mentioned on the abcam data sheet and the lysates were prepared using a non denaturing lysis buffer, aliquoted and stored at -80 degrees centigrade. I use 10% and 12.5% acrylamide gels depending on the molecular weight of the protein. initially 40 ug protein was loaded and the gels were run at 70 volts and transfered on to PVDF membrane (ice cold). All buffers were prepared as suggested in the western blot guide by Abcam. 5% milk was used to block the membrane, washed thrice with TBST and/ or TBS. Incubated with respective antibodies (initially 1: 1000 dilution) overnight in cold room with agitation. The blots were added with secondary antibody the following day (secondary antibody from abcam (ab97023 and ab97051 which were aliquoted and stored at -20 degrees centigrade) incubated for 1 hour at room temperature and then washed thrice with TBS/and or TBST. probed with chemiluminescent solution from Biorad. Bands were not detected. We tried loading 50ug protein (we cannot load more owing to the limited volumes of sample we have and the nature of the project) and then tried using 1:500 and 1:250 dilutions, but still the antibodies were not working. We have been using this protocol and works fine with all the other abcam antibodies. Please let me know if I need to elaborate on any of these steps.

    Read More

    Abcam community

    Verified customer

    Asked on Jan 24 2013

    Answer


    Upon reviewing your protocol, it is troubling to me that all 4 of these antibodies are not working. While it is conceivable that all four antibodies are not performing properly, typically when multiple antibodies are not working it is an issue with sample preparation, membrane transfer, or the detection substrate. In your case, I think the following troubleshooting steps are the most worth your while attempting, with tip 1 being the first thing you try since it is the most likely to make the targets recognizable by the antibodies:
    1. You mentioned your samples were non-denatured. ab2571, ab32124, and ab81212 were all raised against synthetic peptides and will require linear proteins in order to recognize their epitopes. Thus, your samples should be both reduced and denatured (SDS and beta-mercaptoethanol or DTT) and boiled for 5 minutes at 100C prior to freezing at -20C. RIPA buffer is a good choice for mitochondrial and membrane proteins such as these.
    2. To enrich the target proteins, have you considered a mitochondria isolation kit such as ab110170 (https://www.abcam.com/Mitochondria-Isolation-Kit-for-Cultured-Cells-ab110170.html)?
    3. Have you insured that the proteins are being transferred with a reversible stain such as Ponceau S? Since you are using PVDF, make sure to pre-soak the membrane in MeOH, then in transfer buffer.
    4. If you are including milk in your antibody dilutions try removing milk or reducing to 0.5% milk for the antibody dilutions only.
    Further troubleshooting tips, including no signal specific tips, are included through the link below:
    https://www.abcam.com/index.html?pageconfig=resource&rid=11352
    Please let me know the results of troubleshooting, especially step 1. I look forward to your reply and sincerely hope this information helps.

    Read More

    Abcam Scientific Support

    Answered on Jan 24 2013

    Question

    1) Code du produit Abcam ab119684

    2) Numéro de commande Abcam ou le numéro de lot/batch xx Description du problème Bande située à 29Kda au lieu des 20KDa indiqués

    4) Préparation de l’échantillon :

    Type d’échantillon (lysat cellulaire, fraction, protéine recombinante …) :

    Tampon de lyse 50mM TRISHCL / 150mM NaCl / 0.5% NP / 0.25% Sodium Deoxycolate / 0.1% SDS

    Inhibiteurs de protéases 1mM PMSF + Complete mini 1X (Roche 11836153001)

    Inhibiteurs de phosphatases NON

    Agent réductant Laemli 1X final

    95oC pendant ≥5 min? oui/non OUI

    Protéine/puits en mg ou nombre de cellules/puits 25µg

    Contrôle positif NON

    Contrôle négatifxx

    5) Pourcentage du gel Stacking: 4% / Separating: 12%

    Type de membrane PVDF

    Vérification du transfert NON

    Tampon de saturation et concentration Lait 5% en TBS-Twenn

    Temps d’incubation de la saturation 1 Heure

    Température de saturation Température ambiante (environ 24°c)

    6 Anticorps primaire (utilisation de plusieurs anticorps, description dans “notes supplémentaires”) :

    Concentration ou dilution 1/1000

    Tampon diluant TBS-Tween + 0.1% lait

    Temps d’incubation 1 Heure

    Température Incubation Température ambiante (environ 24°c)

    7) Anticorps secondaire :

    Espèce

    Réagit contre Mouse

    Concentration ou dilution 1/15000

    Tampon de dilution TBS-Tween

    Temps d’incubation 1 Heure

    Température d’incubation Température ambiante (environ 24°c)

    Fluorochrome or enzyme conjuguée Peroxydase

    8) Lavage après les anticorps primaire et secondaire :

    Tampon TBS-Tween

    Nombre de lavage 3 fois

    Temps d’incubation du lavage 5 minutes

    9) Méthode de détection: ECL + (Ge Healthcare RPN 2132)

    10) Combien de fois avez vous essayer ce Western Blot ? environ 6 fois

    Obtenez-vous toujours le même résultat ? OUI

    Quelle étape avez vous optimisée ?

    Aucune, car, même si elle n’est pas située au poids apparent attendu, c’est la seule bande apparaissant sur le film…

    Read More

    Abcam community

    Verified customer

    Asked on Apr 10 2012

    Answer

    Merci d'avoir pris le temps de remplir notre questionnaire et de nous avoir contactés. Je suis désolée que vous avez des problèmes avec l'interprétation de deux de nos anticorps.

    Comme expliqué auparavant, notre politique est d'offrir tout d'abord la meilleure assistance technique. Si jamais nous concluons que le produit que vous avez reçu ne fonctionne pas comme mentionné sur sa fiche technique et que ce produit a été commandé sous 180 jours, nous nous ferons un plaisir de vous offrir un remplacement gratuit ou un avoir en compensation.

    J'aimerais donc vous offrir quelques suggestions afin d'optimiser vos résultats. J'apprécierais si vous pouviez confirmer quelques détails de la procédure.

    1.) Comme écrit dans mon premier émail, est-ce que vous avez vérifié que le tampon du migration est compatible/adapté au marquer de poids? Effectivement, suivant le tampon, l'échelle dans le tampon peux varier beaucoup. Pour illustration, je peux vous recommander de voir l'image comparative d'un de nos marqueurs en différents tampons: Click here (or use the following: https://www.abcam.com/Prestained-Protein-Ladder-Mid-range-molecular-weight-10-175-kDa-ab115832.html).

    2.) Il existent deux isoformes du mtTFA. Celles-ci sont décrites dans la base de données SwissProt chez la souris: http://www.uniprot.org/uniprot/P40630 La deuxième isoforme a un poids moléculaire attendu de 25kD et est appelé isoforme nucléaire.

    3.) Je peux vous conseiller de tester des autres cellules, comme les cellules Hela, en parallèle comme test. Ceci afin d'exclure que dans les fibroblastes une nouvelle isoforme soit exprimé.

    4.) Est-ce que vous pouvez confirmer aussi que vous avez utilisé des extrait totaux?

    J'espère que ces informations vous aideront. Si nos suggestions ne permettent pas d'améliorer vos résultats, n'hésitez pas à me recontacter pour plus d'assistance.

    Read More

    Abcam Scientific Support

    Answered on Apr 10 2012

    Question

    Nous avons récemment testé deux références d’anticorps :
    1. Anti-Surf1 antibody [21H2BG4] (ab110256)
    2. Anti-mtTFA antibody [18G102B2E11] (ab119684)
    Pour ces deux références, nous avons bien obtenu une ou plusieurs bande(s). Mais aucune bande prédominante située au poids moléculaire attendu (Voir le .pdf joint)
    Tous nos WB ont été fait à partir d’extraits protéiques issus de Fibroblastes humains.

    En ce qui concerne SURF1, nous somme en possession de cellules de patients mutés dans pour cette protéine. Je vous présente leurs résultats dans le .pdf. Certains d’entre eux voient la bande (au mauvais poids moléculaire) disparaître, mais pas tous…Est-ce bien cette bande ? car il n’y aucune bande au poids moléculaire attendu. Il est dit dans la fiche technique que la quantité de la protéine SURF1 est faible dans les cellules.
    Pour mtTFA, nous n’avons aucun moyen de vérifier que cette bande est bien la bonne. Hormis le fait que cette bande est de loin prédominante sur les autres et que ses variations vont dans le sens de notre hypothèse… ce qui est un peu juste

    Auriez-vous eu quelques échos de personnes travaillant avec ces anticorps sur le même matériel cellulaire, c'est-à-dire fibroblastes humains ?
    Ont-il observé ces mêmes différences ?
    Vous a-t-on fait remarqué ces différences dans d’autres types cellulaire ?

    Read More

    Abcam community

    Verified customer

    Asked on Mar 27 2012

    Answer

    Je suis désolée d'apprendre que ces anticorps ne vous donnent pas de résultats satisfaisants en Western blot - c.à.d. une bande à environ 42kD pour SURF1 et à environ 24kD pour mtTFA).
    Nous n'avons pas eu des autres retours négatives et n'avons pas non plus eu des indications autres d'un problème avec ces anticorps. Nous n'avons pas non plus eu des feedback autres des gens utilisant ces anticorps sur des cellules similaires.
    Comme la qualité de nos produits est très imporante pour nous, nous aimerions bien investiguer en plus de profondeur le problème que vous avez avec ces anticorps. Veuillez pour ceci nous fournir un peu plus d'information s'il vous plaît. Pour ceci je vous ai joint un questionnaire qui nous permet de regrouper le plus d'informations possible sur le protocole que vous avez utilisé avec cet anticorps..
    En regardant les informations que vous nous aviez déjà envoyé, j'aimerai bien noter les points suivants:
    1.) Est-ce que vous avez vérifié que le tampon du migration est compatible/adapté au marquer de poids? Effectivement, suivant le tampon, l'échelle dans le tampon peux varier beaucoup. Pour illustration, je peux vous recommander de voir l'image comparative d'un de nos marqueurs en différents tampons: Click here (or use the following: https://www.abcam.com/Prestained-Protein-Ladder-Mid-range-molecular-weight-10-175-kDa-ab115832.html).
    2.) Il existent deux isoformes du mtTFA. Celles-ci sont décrites dans la base de données SwissProt chez la souris: http://www.uniprot.org/uniprot/P40630 La deuxième isoforme a un poids moléculaire attendu de 25kD et est appelé isoforme nucléaire.
    Je me réjouis de recevoir votre réponse avec les informations additionnelles afin de pouvoir continuer à investiguer ce problème.
    Si nous concluons que notre produit est fautif et ne fonctionne pas comme explicité par notre fiche technique, nous serons ravis de mettre en place l'envoi d'un tube de remplacement gratuit ou d'un remboursement (sous réserve d'une commande ultérieure de 180 jours).

    Read More

    Abcam Scientific Support

    Answered on Mar 27 2012

    Please note: All products are "FOR RESEARCH USE ONLY. NOT FOR USE IN DIAGNOSTIC PROCEDURES"
    For licensing inquiries, please contact partnerships@abcam.com

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