Overview

  • Product name
    Anti-RNA polymerase II CTD repeat YSPTSPS antibody [8WG16] - ChIP Grade
    See all RNA polymerase II CTD repeat YSPTSPS primary antibodies
  • Description
    Mouse monoclonal [8WG16] to RNA polymerase II CTD repeat YSPTSPS - ChIP Grade
  • Host species
    Mouse
  • Tested applications
    Suitable for: CHIPseq, WB, ICC/IF, IHC - Wholemount, ChIP, Flow Cyt, IPmore details
  • Species reactivity
    Reacts with: Mouse, Cow, Human, Saccharomyces cerevisiae, Xenopus laevis, Arabidopsis thaliana, Caenorhabditis elegans, Drosophila melanogaster, Schizosaccharomyces pombe, Quail, Neurospora crassa, Rice
    Predicted to work with: a wide range of other species
  • Immunogen

    Other Immunogen Type corresponding to RNA polymerase II CTD repeat YSPTSPS. Immunogen: Wheat germ RNA Polymerase II

  • Positive control
    • ChIP: Nuclear lysate from Arabidopsis thaliana seedlings; Nuclear lysate of MCF7 cells. WB: HeLa total lysate; HeLa, THP-1 and RAW 264.7 cell lysates. ICC/IF: HeLa cells. IHC-Wm: Caenorhabditis elegans embryo. Flow Cyt: HeLa cells.
  • General notes
    This antibody may be used to detect unproteolyzed RNA polymerase II or to inhibit specific transcription from class II promoters. This is a polyol-responsive antibody and can be used in gentle purifications of RNA polymerase II from from wheat germ, calf thymus, and yeast and is likely to purify RNA polymerase II from most eukaryotic organisms. It inhibits promoter-directed transcription, but it does not inhibit elongation in the nonspecific transcription assay.

    Abcam recommended secondaries - Goat Anti-Mouse HRP (ab205719) and Goat Anti-Mouse Alexa Fluor® 488 (ab150113).

    See other anti-mouse secondary antibodies that can be used with this antibody.

Properties

Applications

Our Abpromise guarantee covers the use of ab817 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
CHIPseq Use at an assay dependent concentration. PubMed: 19251593Use 2ug per 0.3ml of sonicated chromatin.
WB Use a concentration of 0.25 - 2 µg/ml. Detects a band of approximately 217 kDa (predicted molecular weight: 217 kDa).
ICC/IF Use a concentration of 1 - 5 µg/ml.
IHC - Wholemount Use at an assay dependent concentration.

See Abreview.

ChIP Use at an assay dependent concentration.

See Abreview; the user recommends using anti-mouse IgG coated Dynabeads instead of Protein A to recover the precipitate).

Flow Cyt Use 0.5µg for 106 cells.

ab170191 - Mouse monoclonal IgG2a, is suitable for use as an isotype control with this antibody.

We recommend using Goat Anti-Mouse IgG H&L (DyLight® 488) preadsorbed (ab96879) secondary antibody.

IP Use at an assay dependent concentration.

Target

  • Function
    DNA-dependent RNA polymerase catalyzes the transcription of DNA into RNA using the four ribonucleoside triphosphates as substrates. Largest and catalytic component of RNA polymerase II which synthesizes mRNA precursors and many functional non-coding RNAs. Forms the polymerase active center together with the second largest subunit. Pol II is the central component of the basal RNA polymerase II transcription machinery. It is composed of mobile elements that move relative to each other. RPB1 is part of the core element with the central large cleft, the clamp element that moves to open and close the cleft and the jaws that are thought to grab the incoming DNA template. At the start of transcription, a single-stranded DNA template strand of the promoter is positioned within the central active site cleft of Pol II. A bridging helix emanates from RPB1 and crosses the cleft near the catalytic site and is thought to promote translocation of Pol II by acting as a ratchet that moves the RNA-DNA hybrid through the active site by switching from straight to bent conformations at each step of nucleotide addition. During transcription elongation, Pol II moves on the template as the transcript elongates. Elongation is influenced by the phosphorylation status of the C-terminal domain (CTD) of Pol II largest subunit (RPB1), which serves as a platform for assembly of factors that regulate transcription initiation, elongation, termination and mRNA processing. Acts as an RNA-dependent RNA polymerase when associated with small delta antigen of Hepatitis delta virus, acting both as a replicate and transcriptase for the viral RNA circular genome.
  • Sequence similarities
    Belongs to the RNA polymerase beta' chain family.
  • Domain
    The C-terminal domain (CTD) serves as a platform for assembly of factors that regulate transcription initiation, elongation, termination and mRNA processing.
  • Post-translational
    modifications
    The tandem heptapeptide repeats in the C-terminal domain (CTD) can be highly phosphorylated. The phosphorylation activates Pol II. Phosphorylation occurs mainly at residues 'Ser-2' and 'Ser-5' of the heptapeptide repeat and is mediated, at least, by CDK7 and CDK9. CDK7 phosphorylation of POLR2A associated with DNA promotes transcription initiation by triggering dissociation from DNA. Phosphorylation also takes place at 'Ser-7' of the heptapeptide repeat, which is required for efficient transcription of snRNA genes and processing of the transcripts. The phosphorylation state is believed to result from the balanced action of site-specific CTD kinases and phosphatases, and a 'CTD code' that specifies the position of Pol II within the transcription cycle has been proposed. Dephosphorylated by the protein phosphatase CTDSP1.
    Among tandem heptapeptide repeats of the C-terminal domain (CTD) some do not match the Y-S-P-T-S-P-S consensus, the seventh serine residue 'Ser-7' being replaced by a lysine. 'Lys-7' in these non-consensus heptapeptide repeats can be alternatively acetylated, methylated and dimethylated. EP300 is one of the enzyme able to acetylate 'Lys-7'. Acetylation at 'Lys-7' of non-consensus heptapeptide repeats is associated with 'Ser-2' phosphorylation and active transcription. It may regulate initiation or early elongation steps of transcription specially for inducible genes.
    Methylated at Arg-1810 prior to transcription initiation when the CTD is hypophosphorylated, phosphorylation at Ser-1805 and Ser-1808 preventing this methylation. Symmetrically or asymmetrically dimethylated at Arg-1810 by PRMT5 and CARM1 respectively. Symmetric or asymmetric dimethylation modulates interactions with CTD-binding proteins like SMN1/SMN2 and TDRD3. SMN1/SMN2 interacts preferentially with the symmetrically dimethylated form while TDRD3 interacts with the asymmetric form. Through the recruitment of SMN1/SMN2, symmetric dimethylation is required for resolving RNA-DNA hybrids created by RNA polymerase II, that form R-loop in transcription terminal regions, an important step in proper transcription termination. CTD dimethylation may also facilitate the expression of select RNAs. Among tandem heptapeptide repeats of the C-terminal domain (CTD) some do not match the Y-S-P-T-S-P-S consensus, the seventh serine residue 'Ser-7' being replaced by a lysine. 'Lys-7' in these non-consensus heptapeptide repeats can be alternatively acetylated, methylated and dimethylated. Methylation occurs in the earliest transcription stages and precedes or is concomitant to 'Ser-5' and 'Ser-7' phosphorylation.
    Ubiquitinated by WWP2 leading to proteasomal degradation (By similarity). Following UV treatment, the elongating form of RNA polymerase II (RNA pol IIo) is ubiquitinated UV damage sites without leading to degradation: ubiquitination is facilitated by KIAA1530/UVSSA and promotes RNA pol IIo backtracking to allow access to the nucleotide excision repair machinery.
  • Cellular localization
    Nucleus.
  • Information by UniProt
  • Database links
    see all
  • Alternative names
    • DNA directed RNA polymerase II A antibody
    • DNA-directed RNA polymerase II largest subunit RNA polymerase II 220 kd subunit antibody
    • DNA-directed RNA polymerase II subunit A antibody
    • DNA-directed RNA polymerase II subunit RPB1 antibody
    • DNA-directed RNA polymerase III largest subunit antibody
    • hRPB220 antibody
    • hsRPB1 antibody
    • POLR2 antibody
    • Polr2a antibody
    • POLRA antibody
    • Polymerase (RNA) II (DNA directed) polypeptide A 220kDa antibody
    • Polymerase (RNA) II (DNA directed) polypeptide A antibody
    • RNA polymerase II subunit B1 antibody
    • RNA-directed RNA polymerase II subunit RPB1 antibody
    • RPB1 antibody
    • RPB1_HUMAN antibody
    • RPBh1 antibody
    • RpIILS antibody
    • RPO2 antibody
    • RPOL2 antibody
    see all

Images

  • Various regions across the Actin2/7 loci were tested for the presence of RNA polymerase II CTD repeat YSPTSPS. A nuclear lysate from Arabidopsis thaliana seedlings was crosslinked using 1% formaldehyde for 30 seconds.  The ChIP was performed with 0.1 µg of ab817 per µg of chromatin; incubated together for 16 hours at 4°C.  The immunoprecipitated DNA was quantified by Real-Time PCR.  The bottom panel indicates the positive (ab817) and negative controls (no antibody) at region B3.

    See Abreview

  • Anti-RNA polymerase II CTD repeat YSPTSPS antibody [8WG16] - ChIP Grade (ab817) at 1/500 dilution + HeLa (human epithelial cell line from cervix adenocarcinoma) total lysate at 15 µg

    Secondary
    Goat anti-mouse-IgG secondary antibody conjugated to HRP at 1/3000 dilution

    Predicted band size: 217 kDa

  • 2% PFA-fixed, 0.5% Triton X-100 permeabilized HeLa (human epithelial cell line from cervix adenocarcinoma) cells stained for RNA polymerase II CTD repeat YSPTSPS using ab817 (red) at 2 µg/ml in ICC/IF. Secondary antibody: DyLight™ 594 conjugated goat anti-mouse IgG for 1 hour at RT. Actin filaments were labeled with Alexa Fluor® 488 Phalloidin (green). Nuclei were counterstained with DAPI (blue).

  • IHC - Wholemount of Caenorhabditis elegans embryo labelling RNA polymerase II CTD repeat YSPTSPS with ab817. Sample was incubated with primary antibody (1/100 in PBS + 3% BSA + 0.1% Triton-X 100) for 24 hours at 4°C. ab150113, a goat anti-mouse Alexa Fluor® 488 (undiluted) was used as the secondary antibody.

    See Abreview

  • Overlay histogram showing HeLa (human epithelial cell line from cervix adenocarcinoma) cells stained with ab817 (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 (ab817, 0.5µg/1x106 cells) for 30 min at 22ºC. The secondary antibody used was a Goat Anti-Mouse IgG H&L (DyLight® 488) preadsorbed (ab96879) at 1/500 dilution for 30 min at 22ºC. Isotype control antibody (black line) was mouse IgG2a [ICIGG2A] (ab91361, 1µg/1x106 cells) used under the same conditions. Acquisition of >5,000 events was performed.

  • All lanes : Anti-RNA polymerase II CTD repeat YSPTSPS antibody [8WG16] - ChIP Grade (ab817) at 0.25 µg/ml

    Lane 1 : HeLa (human epithelial cell line from cervix adenocarcinoma) cell lysate
    Lane 2 : THP-1 (human monocytic leukemia cell line) cell lysate
    Lane 3 : RAW 264.7 (mouse macrophage cell line transformed with Abelson murine leukemia virus) cell lysate

    Lysates/proteins at 15 µg per lane.

    Developed using the ECL technique.

    Predicted band size: 217 kDa

  • Chromatin was prepared from nuclear lysate of the human MCF7 breast epithelial adenocarcinoma cells. The cross-linking (X-ChiP) technique was used, crosslinking was performed for 15 minutes in formaldehyde. The primary antibody was used in concentration of 0.2 µg/µg chromatin and incubated with the sample for 16 hours at 4°C in SDS, DOC, TritonX-100, EDTA, HEPES, NaCl. The immunoprecipitated DNA was quantified by real time PCR. Ct values were converted to DNA copy numbers using a standard curve in the Q-PCR step. The number of binding events detected for each test reaction was then calculated by taking into account the DNA copy number, cell equivalents of chromatin used in the ChIP and PCR, and primer pair amplification efficiency.

    See Abreview

References

This product has been referenced in:
  • Miao Y  et al. Enhancer-associated long non-coding RNA LEENE regulates endothelial nitric oxide synthase and endothelial function. Nat Commun 9:292 (2018). Read more (PubMed: 29348663) »
  • Mondal T  et al. Two way controls of apoptotic regulators consign DmArgonaute-1 a better clasp on it. PLoS One 13:e0190548 (2018). Read more (PubMed: 29385168) »
See all 212 Publications for this product

Customer reviews and Q&As

1-10 of 21 Q&A

Answer

The earliest reference I found for the clone is:

J Biol Chem. 1989 Jul 5;264(19):11511-20.
PMID:2472398
http://www.jbc.org/content/264/19/11511.long

It describes production of the clone and some characterization but as far as I can tell, they do not assess affinity for phosphorylated Pol II CTD.

A search on PubMed using the clone ID as a search term returns papers that have more characterization data, including this one:

Nucleic Acids Res. 1996 Dec 1;24(23):4649-52.
PMID:8972849
http://www.ncbi.nlm.nih.gov/pubmed/8972849

But a more relevant paper may be the one that is mentioned in a reply to a similar question we received in May 2007, in the "Q&A" section of the datasheet, https://www.abcam.com/RNA-polymerase-II-CTD-repeat-YSPTSPS-antibody-8WG16-ChIP-Grade-ab817/questions/20092 .

J Biol Chem. 2005 Sep 9;280(36):31368-77.
PMID: 16012166
http://www.ncbi.nlm.nih.gov/pubmed/?term=16012166

From the Results:

"...Of the antibodies directed at the CTD, 8WG16 binds the unphosphorylated consensus repeat, YSPTSPS, whereas H14 requires phosphorylation at Ser-5 (32, 45–47)...Unsurprisingly, the H14 antibody precipitated Pol II with the H14 epitope most effectively, but each of the antibodies precipitated some phosphorylated Pol II (Fig. 1A, compare B lanes).."

A paragraph or two later, there is "...data that suggest that the unphosphorylated Ser-2 is required for the 8WG16 epitope."

I hope this helps. Please let us know if you still have questions.

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Answer

I've compiled some information below regarding potential primer sets for the ChIP antibodies we talked about, where information was lacking (verified in ChIP by other researches), I included links to abReviews and references that used the antibody in ChIP.

1. ab817: GAPDH (+), GAD1 (-), AFM (-), Factor VIII (-)

2. ab1791: GAPDH, RPL30, ALDOA, MYO-D, SERPINA, AFM, SAT2, SATa (each batch is verified against this primer set, an example image of one batch test is given on the datasheet)

3. ab7030: was verified in ChIP by other researchers submitting data through our abReview system and specific references. Their positive and negative controls used are given through the link below, as well as a "click to contact this user" link that allows you to contact the researchers if you would like more specific primer information. The specific references with ChIP listed in bold are good sources for positive and negative primers as well.
a) Abreview: (NFKBIA promoter (+), GAPDH (-)) https://www.abcam.com/HDAC3-antibody-ChIP-Grade-ab7030-reviews.html?intabreviewid=25057
b) Abreview: https://www.abcam.com/HDAC3-antibody-ChIP-Grade-ab7030-reviews.html?intabreviewid=18386
c) Specific references: https://www.abcam.com/HDAC3-antibody-ChIP-Grade-ab7030-references.html

4. ab111069 Untested in ChIP.

5. ab992: was verified in ChIP by other researchers (abReviews and specific references)
a) Specific references: https://www.abcam.com/Rad21-antibody-ChIP-Grade-ab992-references.html
b) Abreview: https://www.abcam.com/Rad21-antibody-ChIP-Grade-ab992-reviews.html?intabreviewid=14542
c) Abreview: (EBV 3+ (+), EBV1,2,4,5,6 (-), GAPDH (-), Cen (-)) https://www.abcam.com/Rad21-antibody-ChIP-Grade-ab992-reviews.html?intabreviewid=33942

Our ChIP primer sets:
https://www.abcam.com/index.html?pageconfig=resource&rid=10600

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Answer

Merci de nous avoir contactés.

Le numéro de commande pour ab26721 et ab1791 est xxxxxxxx. Vous recevrez prochainement un mail de confirmation comprenant les détails d'expédition.

Le numéro de commande pour ab106293 est maintenant xxxxxxxxx. Cet anticorps vous sera envoyé quand il sera en stock.

N'hésitez pas à nous contacter lors d'une prochaine occasion.

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Answer

Merci de nous avoir contactés.

Nous sommes désolés d'apprendre que les produits que vous avez reçus ne fonctionnent pas comme attendu. Je me permets de vous transférer le numéro de commande des remplacements gratuits de ces produits : xxxxxxxxxx

Je dois vous informer qu’ab106293 n’est pas en stock et nous le recevrons dans 2-3 semaines. Si vous préférez recevoir ab26721 et ab1791 (qui sont en stock) avant cette date, s’il vous plait contactez le service clientèle sur :

mailto:orders@abcam.com
Tel: 01.46.94.62.96
Fax: 01.70.80.96.08

en citant ce numéro de commande.

Vous recevrez prochainement un mail de confirmation comprenant les détails d'expédition.

N'hésitez pas à nous contacter lors d'une prochaine occasion.

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Answer

Merci de nous avoir contactés.

Nous sommes désolés d'apprendre que les produits que vous avez reçus ne fonctionnent pas comme attendu. Je suis d’accord pour vous envoyer ab26721, ab106293 et ab1791 en remplacement d’ab817, ab5408 et ab24758 qui n’ont pas fonctionnés comme indiqué sur la fiche technique.

Pour faire ces remplacements, pouvez-vous m’envoyer le numéro de commande original? J’ai trouvé la commande pour ab817 (commande xxxxxxxxx.) Par contre je n’ai pas trouvé la commande placé pour ab5408 et ab24758 pouvez-vous me faire parvenir ce numéro de commande ou le nom de la personne qui a fait passer la commande ?

J’attends votre réponse.

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Question

carlesc@ujf-grenoble.fr a écrit :
> Bonjour,
>
> Je vous contacte suite à des tests que nous avons effectués pour
> vérifier la qualité des Anticorps que nous avons commandé chez vous.
> Certains d'entre-eux ne reconnaissent pas les antigènes attendus, ou
> mal, ou n'ont pas la spécificité mentionnée sur les fiches produit.
> Je joins en attachement le document support avec les résultats
> d'expériences de dot-blot réalisées par la post-doctorante qui
> travaille avec moi.
> Vous trouverez aussi en attachement le Proforma correspondant à
> l?ensemble des peptides que nous avons commandés auprès de chez vous
> pour tester les anticorps.
>
> Ci-dessous, le bilan de dysfonctionnement des Anticorps Abcam :
>
> - ab817 against unphosphorylated form of CTD ; produces very low/no signal.
> - ab5408 should be against all, but "ELISA and peptide blocking show
> that the specificity of ab5408 towards the different phospho-forms
> varies between batches, with a preference towards the phosphoS5
> form" ; binds only S5P in our experiment.
> - ab24758 should be against S2P ; does not work at all.
>
> Pour poursuivre nos travaux, nous aurions besoin de votre part (en
> ChIPseq-grade):
> 1) d?un anticorps anti-CTD non-phosphorylé : dans votre catalogue,
> vous recommendez ab26721 (« another suitable antibody that shows a
> greater preference for unmodified RNAPII in WB peptide inhibition."
> ). Est-il fiable pour une reconaissance quasi exclusive de CTD non
> phosphorylé ?
> 2) d?un anticorps anti-CTD toutes isoformes
>
> De plus, nous recherchons aussi :
> 3) un anticorps anti-Histone 3:
> ab1791 est-il fiable pour les expériences de ChIPseq ?
>
> Ayant perdu beaucoup de temps à des expériences de ChIP et dépensé
> énormément d?argent, également pour leur trouble-shooting (achat de
> tous les peptides, très honéreux), nous espérons vivement un geste
> gracieux de votre part.

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Answer

Merci de nous avoir contactés.

J’ai eu ma réponse de nos laboratoires et ce cas et assez compliqué. Premièrement je tiens à vous remercier d’avoir patienté et j’apprécie énormément l’effort et le temps que vous avez passé avec ces anticorps.

Je peux maintenant vous expliquer ou nous en sommes avec ces anticorps puisque il y a eu des fautes avec les fiches techniques.

Pour les anticorps qui ne fonctionne pas :

Ab817 (pas de signal): Nous n’avons pas testé cet anticorps contre la CTD non-phosphorylé et la CTD phosphorylé donc nous ne pouvons pas garantir une spécificité contre la CTD non-phosphorylé. Avant nous avions une image qui suggérait que cet anticorps reconnaissait seulement la CTD non-phosphorylé. Cette image a maintenant été supprimée parce que c’était une erreur sur notre fiche technique et nous sommes désolés pour cela.

Sur votre dot blot il n’y a aucun dot donc je conclue que ce produit n’a pas fonctionné selon la fiche technique et je vous propose un remplacement gratuit ou un avoir ou remboursement en compensation.

Ab5408 (un dot pour ser5 uniquement) : l’immunogène de cet anticorps est “10 repeats of synthetic peptide YSPTSPS using chemically synthesized phospho-ser 5” donc cet anticorps devrait reconnaitre phospho-ser 5 uniquement comme indique vos résultats. Par contre puisque il y avait une erreur sur la fiche technique, je vous propose également un remplacement gratuit ou un avoir ou remboursement en compensation.

Ab24758 (pas de marquage) : cet anticorps aurait dû donner un dot pour ser2. Sur votre dot blot il n’y a aucun dot donc je conclue que ce produit n’a pas fonctionné selon la fiche technique et je vous propose un remplacement gratuit ou un avoir ou remboursement en compensation.



En tout, cela vous fait 3 anticorps pour lesquels je peux vous faire un remplacement ou en avoir.

En ce qui concerne les produits que vous recherchiez je peux vous proposer les suivants :

Ab26721. J’ai contacté le laboratoire qui m’a confirmé que le lot que nous avons en stock ne reconnait pas phosphoS2 ni phosphoS5 et donc reconnait uniquement la protéine non-phosphorylé en WB. Nos manipulations montre que ce lot reconnait <2% les formes phosphorylé.

Ab1797 est garanti pour fonctionner en ChIPseq.

Pour un anticorps anti-CTD toutes isoformes nous en n’avons pas en stock qui soit « ChIP grade.» Nous avons un anticorps ab106293 (www.abcam.com/ab106293) qui n’est pas ChIP grade mais qui devrait reconnaitre toutes les isoformes. L’immunogène c’est » Recombinant fragment, corresponding to amino acids 195-422 of Human RNA polymerase II CTD repeat YSPTSPS (BC067295). » Si vous souhaitez le tester en ChIP. Il me serait possible de vous faire une offre de test cependant, vous serez obligé de l’acheter dans un premier temps pour que cette offre soit valable.



Étapes à suivre :

1. Nous informer que vous souhaitez tester cet anticorps en ChIP et nous vous enverrons un code promotionnel (valable 4 mois) à utiliser après obtention et envoi de vos résultats,

2. Acheter l'anticorps,

3. Nous envoyer vos résultats (positifs ou négatifs) obtenus sous forme d'Abreview (www.abcam.com/abreviews).

4. L'envoi de vos résultats activera le code et vous donnera un avoir de la valeur de l'anticorps testé.

J'espère que ces informations vous aideront. J’attends votre réponse en ce qui concerne comment vous aimeriez procéder avec ce cas.

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Question
Answer

Thank you for your reply and for kindly confirming these details.

I am sorry to confirm we do not have isotype control antibodies to match the primary antibodies you mention that have been tested in ChIP or ChIP-seq. We do have the following which I can recommend to consider trying, but please note these are regrettably not covered by the guarantee for ChIP or ChIP-seq.

ab817 is a mouse IgG2a primary. The suitable isotype control would be:

ab91361 Mouse IgG2a [ICIGG2A] - Isotype Control
Tested in: Flow cytometry
https://www.abcam.com/index.html?datasheet=91361 (or use the following: https://www.abcam.com/index.html?datasheet=91361).

ab5408 is a mouse mouse IgG1 primary . The suitable isotype control would be:

ab81216 Mouse IgG1 [CT6] - Isotype control
Flow Cyt, WB, IHC-P
https://www.abcam.com/index.html?datasheet=5408 (or use the following: https://www.abcam.com/index.html?datasheet=5408).

I hope this will be helpful. Please do not hesitate to let me know if you have any further questions.

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Question
Answer

Merci de nous avoir contactés.

J'ai compris que vous rencontrez des difficultés avec un de nos produits et j'aimerais essayer de résoudre avec vous ce problème.
Nos produits sont garantis 6 mois pour fonctionner tels qu'ils sont décrits sur la fiche technique (garantie Abpromise), nous pouvons donc toujours résoudre le problème que vous avez rencontré en vous apportant un support technique, vous rembourser, vous faire un avoir, ou vous envoyer un anticorps de remplacement.

Pourriez-vous, s'il vous plait me communiquer et/ou confimer les informations suivantes :
- la référence de l'anticorps. ab817?
- le numéro de lot. 610713?
- le numéro et la date de la commande
- l'application testée. ChIP?
- le type d'échantillon (espèce, lignée cellulaire, tissu)
- quelques détails sur votre protocole (concentration/dilution de l'anticorps, temps d'incubation, agent bloquant, préparation des échantillons, etc.)

Je serai ravi de vous apporter une solution, scientifique ou commerciale, à votre problème.

En attendant votre réponse, je vous souhaite une bonne continuation dans vos recherches.

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Answer

I am very pleased to hear you would like to accept our offer and test ab24342 and ab817 in Sencio. These code will give you the value stated above off your next order before the expiration date. To redeem this offer, please submit an Abreview for Scencio and include this code in the “Additional Comments” section so we know the Abreview is for this promotion. Please remember that submission of the Abreview is sufficient for the discount code to become active.

For more information on how to submit an Abreview, please visit the site:

www.abcam.com/Abreviews

Remember, we publish both positive and negative Abreviews on our datasheets so please submit the results of your tests. The code will be active once the Abreview has been submitted and can be redeemed in one of the following ways:

1) Call to place your order and mention the code to our customer service department;

2) Include the code in your fax order;

3) Place your order on the web and enter the promotional code.

Any feedback that you can provide will be greatly appreciated, whether positive or negative. If you have any further questions, please do not hesitate to contact us. We look forward to receiving your Abreview and wish you luck with your research.

The terms and conditions applicable to this offer can be found here:

www.abcam.com/abtrial

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Answer

Thank you for your reply.

I can confirm the alignment of the TBP antibody ab24342 with the Senecio TBP sequence you provided is 88%. Therefore, I would suggest that ab24342 would be likely to detect the Senecio TBP protein.

The alignment of the immunogen of TBP antibody ab12089 with the Senecio TBP you provided is low at 30%. Therefore, I would not expect this antibody to detect the Senecio TBP protein.

The immunogen for ab817 RNA POL II antibody has 100% alignment with the Senecio sequence you provided. This antibody should therefore detect the Senecio protein.

Please note that even when alignment with the immunogen is high, we are not able to guarantee an antibody will work in an untested species without further testing

Therefore, I can offer a discount off a future purchase if you buy ab817 RNA POL II antibody and/ or TBP antibody ab24342 now, test them in Senecio and submit feedback to us in the form of an Abreview. It doesn’t matter whether the Abreview is positive or negative, we would just really like to receive your feedback. The discount would represent the value of this product.

If you are interested in this offer, please follow these steps:

1. Reply to this e-mail to let me know that you would like to proceed and test ab817 RNA POL II antibody and/ or TBP antibody ab24342 in Senecio. I will then send a discount code, one code for each antibody. This code must be issued before purchasing the antibodies so please wait for my reply before ordering.

2. Purchase the antibodies either by phone, fax, or online (www.abcam.com).

3. Test it in Senecio

4. Let us know the results, positive or negative, using our Abreview system (this will take about 10 minutes and images are great if you have them!). To find out how to submit an Abreview, please visit: https://www.abcam.com/abreviews.

5. After the review is submitted to us, the discount codes become active. Simply place your new order by phone, fax, or on the web and mention the discount code. The discount can be redeemed for any order and the discount code is valid for 4 months after issue.

Please remember that submission of the Abreview is sufficient for the discount code to become active.

We are always pleased to obtain feedback about our products and any information is greatly appreciated! Even if ab817 RNA POL II antibody and/ or TBP antibody ab24342 turn out to be unsuitable for Senecio, you will still receive the discount on your next purchase after your Abreview has been submitted.

Please let me know if you have any questions about this offer and I would be happy to help you further.

The Terms and Conditions of this offer can be found at: www.abcam.com/abtrial.

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1-10 of 21 Q&A

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