Recombinant
RabMAb

Recombinant Anti-MUC16 antibody [EPSISR23] (ab134093)

Rabbit recombinant monoclonal MUC16 antibody [EPSISR23]. Validated in WB, Flow Cyt, ICC/IF and tested in Human.

Overview

  • Product name

    Anti-MUC16 antibody [EPSISR23]
    See all MUC16 primary antibodies
  • Description

    Rabbit monoclonal [EPSISR23] to MUC16
  • Host species

    Rabbit
  • Tested applications

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

    Reacts with: Human
  • Immunogen

    Synthetic peptide corresponding to Human MUC16 aa 12450-12550.
    Database link: Q8WX17
    (Peptide available as ab139976)

  • Positive control

    • Human ovary cancer, Human uterus and NIH:OVCAR-3 cell lysates; OVCAR-3 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.

     

    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 rabbit monoclonal antibody.

Properties

Applications

Our Abpromise guarantee covers the use of ab134093 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 1/10000 - 1/50000. Predicted molecular weight: 1519 kDa.Can be blocked with MUC16 peptide (ab139976).
Flow Cyt 1/100 - 1/1000.

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

 

ICC/IF 1/100 - 1/250.
  • Application notes
    Is unsuitable for IHC-P or IP.
  • Target

    • Function

      Thought to provide a protective, lubricating barrier against particles and infectious agents at mucosal surfaces.
    • Tissue specificity

      Expressed in corneal and conjunctival epithelia (at protein level). Overexpressed in ovarian carcinomas and ovarian low malignant potential (LMP) tumors as compared to the expression in normal ovarian tissue and ovarian adenomas.
    • Sequence similarities

      Contains 2 ANK repeats.
      Contains 56 SEA domains.
    • Domain

      Composed of three domains, a Ser-, Thr-rich N-terminal domain, a repeated domain containing more than 60 partially conserved tandem repeats of 156 amino acids each (AAs 12061-21862) and a C-terminal transmembrane contain domain with a short cytoplasmic tail.
    • Post-translational
      modifications

      Heavily O-glycosylated; expresses both type 1 and type 2 core glycans.
      Heavily N-glycosylated; expresses primarily high mannose and complex bisecting type N-linked glycans.
      May be phosphorylated. Phosphorylation of the intracellular C-terminal domain may induce proteolytic cleavage and the liberation of the extracellular domain into the extracellular space.
      May contain numerous disulfide bridges. Association of several molecules of the secreted form may occur through interchain disulfide bridges providing an extraordinarily large gel-like matrix in the extracellular space or in the lumen of secretory ducts.
    • Cellular localization

      Cell membrane. Secreted > extracellular space. May be liberated into the extracellular space following the phosphorylation of the intracellular C-terminus which induces the proteolytic cleavage and liberation of the extracellular domain.
    • Information by UniProt
    • Database links

    • Alternative names

      • CA 125 antibody
      • CA-125 antibody
      • CA125 antibody
      • CA125 ovarian cancer antigen antibody
      • Cancer antigen 125 antibody
      • FLJ14303 antibody
      • MUC 16 antibody
      • MUC-16 antibody
      • MUC16 antibody
      • MUC16_HUMAN antibody
      • Mucin 16 antibody
      • Mucin 16 cell surface associated antibody
      • Mucin-16 antibody
      • Mucin16 antibody
      • Ovarian cancer related tumor marker CA125 antibody
      • Ovarian cancer-related tumor marker CA125 antibody
      • Ovarian carcinoma antigen CA125 antibody
      see all

    Images

    • Overlay histogram showing Hela cells stained with ab134093 (red line). The cells were fixed with 80% methanol (5 min) and then permeabilized with 0.1% PBS-Tween for 20 min. The cells were then incubated in 1x PBS / 10% normal goat serum / 0.3M glycine to block non-specific protein-protein interactions followed by the antibody (ab134093, 1/100 dilution) for 30 min at 22°C. The secondary antibody used was Alexa Fluor® 488 goat anti-rabbit IgG (H&L) (ab150077) at 1/2000 dilution for 30 min at 22°C. Isotype control antibody (black line) was rabbit IgG (monoclonal) (1μg/1x106 cells) used under the same conditions. Unlabelled sample (blue line) was also used as a control. Acquisition of >5,000 events were collected using a 20mW Argon ion laser (488nm) and 525/30 bandpass filter. This antibody gave a positive signal in HeLa cells fixed with 4% paraformaldehyde (10 min)/permeabilized with 0.1% PBS-Tween for 20 min used under the same conditions.
    • Anti-MUC16 antibody [EPSISR23] (ab134093) at 1/200000 dilution + NIH: OVCAR-3 (Human ovary adenocarcinoma epithelial cell) whole cell lysate at 20 µg with 5% NFDM/TBST

      Secondary
      Goat Anti-Rabbit IgG (HRP), specific to the non-reduced form of IgG at 1/2000 dilution

      Predicted band size: 1519 kDa

    • All lanes : Anti-MUC16 antibody [EPSISR23] (ab134093) at 1/10000 dilution

      Lane 1 : Human ovary cancer lysate
      Lane 2 : Human uteruslysate
      Lane 3 : NIH:OVCAR-3 cell lysate

      Lysates/proteins at 10 µg per lane.

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

      Predicted band size: 1519 kDa

    • Immunofluorescent analysis of OVCAR-3 cells labelling MUC16 with ab134093 at 1/100 dilution.

    References

    ab134093 has not yet been referenced specifically in any publications.

    Customer reviews and Q&As

    1-5 of 5 Abreviews or Q&A

    Answer

    Thank you for your reply and for clarifying the questions.

    I can confirm that both these antibodies (ab693 and ab134093) are tested and covered by our 6 month guarantee for flow cytometry.

    However, I am sorry they have not been tried and tested for FACS cell sorting where live cells are required, so regrettably we would not be able to guarantee how well this will work. Both contain low concentration of preservative which may affect the cell viability.

    I hope this will be helpful. If you have any further questions, please do not hesitate to contact me.

    Read More

    Answer

    Merci pour ces précisions.

    J'aimerais vous informer que nous n'avons aucun accord commercial avec LuBioScience et que ce revendeur n'est pas en droit de distribuer nos produits. La garantie Abpromise d'Abcam ne s'applique que sur les produits commandés directement chez Abcam ou via un distributeur agréé (https://www.abcam.com/abpromise).

    Je m'engage cependant à faire exception pour le cas présent et accepte de vous envoyer gratuitement une unité de ab14879 directement à votre laboratoire comme convenu précédemment. Le numéro de la commande est ******.

    Pourriez-vous, s'il vous plait, placer vos futures commandes directement chez Abcam et, si possible, informer vos collègues que la société LuBioScience n'est pas autorisée à revendre les produits Abcam.

    N'hésitez pas à m'informer des nouveaux résultats obtenus avec l'anticorps ab134093 et le lysat ab14879.

    Read More

    Question

    Bonjour,
    comme convenu, voici le questionnaire rempli ainsi qu'une phooto de mon blot, le problème étant que les controle positifs OVCAR3 et CA125 recombinant protein ne sont pas reconnu par l'anticorps:
    WB Questionnaire
    1) Abcam product code ab134093
    2) Abcam order reference number or product batch number: je ne l'ai pas à disposition, mais au nom de l'institut de police scientifique à Lausanne.
    3) Description of the problem : non reconnaissance de l’anticorps pour le lysat cellualire OVCAR3 qui m’ a été vendu par Novus comme étant un contrôle positif, et pour la protéine recombinante CA125 Partial Recombinant Protein de novus.
    4) Sample preparation:
    Type of sample (whole cell lysates, fraction, recombinant protein…): lysat celleulaire OVCAR3 et protéine recombinante CA125
    Lysis buffer: aucun
    Protease inhibitors: aucun
    Phosphatase inhibitors: aucun
    Reducing agent: Laemmli+mercaptoethanol
    Boiling for ≥5 min? yes
    Protein loaded ug/lane or cells/lane: 10ug
    Positive control: il s’agissait du controle positif
    Negative control: oui
    5) Percentage of gel: SDS-PAGE 7.5% precast, Biorad
    Type of membrane: PVDF-FL, 0.45u
    Protein transfer verified
    Blocking agent and concentration: Blck de Millipore
    Blocking time: 1 heure
    Blocking temperature: ambiante
    6) Primary antibody (If more than one was used, describe in
    “additional notes”):
    Concentration or dilution: 1/10000
    Diluent buffer: Blck de Millipore
    Incubation time: 1 heure
    Incubation temperature: ambiante
    7) Secondary antibody: Alexa Fluor 532 Goat anti-rabbit IgG
    Species: goat
    Reacts against: rabbit
    Concentration or dilution: 1/1000
    Diluent buffer: Blck de Millipore
    Incubation time: 1 heure
    Incubation temperature: ambiante
    Fluorochrome or enzyme conjugate: Alexa fluor 532
    8) Washing after primary and secondary antibodies: solution de lavage du WesternBreeze de life technologies
    Buffer: PBS
    Number of washes: 3x5min avec solution de lavage et 2x5min
    avec PBS ou eau
    9) Detection method: Laser 532 nm
    10) How many times have you run this staining? 3 fois
    Do you obtain the same results every time? oui

    Read More
    Answer

    Merci pour le questionnaire.

    Nous n'avons pour l'instant en stock que le lot GR93682 pour l'anti-MUC16 ab134093.
    J'ai été informé par mes collègues du laboratoire que le peptide bloquant est disponible à la vente mais que la production de celui-ci prendrait quelques semaines.

    Afin de comprendre et déterminer l'origine du problème rencontré, je peux vous proposer de mettre en place l'envoi d'une unité de lysat nucléaire NIH OVCAR-3 référence ab14879 (https://www.abcam.com/ab14879). Ce lysat a été utilisé comme contrôle positif pour l'anti-MUC16 ab134093 dilué à 1/50,000 en conditions dénaturantes.
    Alternativement je peux vous envoyer une unité d'un anti-MUC16 monoclonal de souris tel que le ab1107 (https://www.abcam.com/ab1107) ou le ab10033 (https://www.abcam.com/ab10033).

    Merci de m'indiquer la résolution qui vous conviendrait le mieux.

    Read More

    Question
    Answer

    Merci pour votre appel. Je suis très désolée que nous n'avions pas répondu après votre premier appel - je ne sais pas ce qui c'est passé.

    Veuillez trouver ci-jointle questionnaire que nous avons discutéau téléphonequi nous permet de regrouper le plus d'informations possible sur le protocole que vous avez utilisé avec cet anticorps.
    Celui-ci nous permettra de vous fournir la meilleure assistance technique.

    Si nous concluons que notre produit est fautif, nous allons vous le remplacer.

    Comme discuté au téléphone, je vais aussi contacter le laboratoire pour l'information des conditions WB ainsi que pour le peptide.

    Je vous remercie par avance de nous envoyer votre protocole.

    Read More

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