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    products/primary-antibodies/s6k1-antibody-e175-ab32359.pdf

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Cell Biology Cell Cycle Cell Cycle Inhibitors Rb
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Validated using a knockout cell lineRecombinantRabMAb

Recombinant Anti-S6K1 antibody [E175] (ab32359)

  • Datasheet
  • SDS
Reviews (4)Q&A (36)References (51)

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Western blot - Anti-S6K1 antibody [E175] (ab32359)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-S6K1 antibody [E175] (ab32359)
  • Western blot - Anti-S6K1 antibody [E175] (ab32359)
  • Anti-S6K1 antibody [E175] (ab32359)

Key features and details

  • Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
  • Rabbit monoclonal [E175] to S6K1
  • Suitable for: WB, IHC-P
  • Knockout validated
  • Reacts with: Mouse, Human

Conjugates logo Related conjugates and formulations

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Overview

  • Product name

    Anti-S6K1 antibody [E175]
    See all S6K1 primary antibodies
  • Description

    Rabbit monoclonal [E175] to S6K1
  • Host species

    Rabbit
  • Specificity

    ab32359 may detect both phosphorylated and non-phosphorylated forms of p70 S6 Kinase.
  • Tested applications

    Suitable for: WB, IHC-Pmore details
    Unsuitable for: Flow Cyt or ICC/IF
  • Species reactivity

    Reacts with: Mouse, Human
    Predicted to work with: Rat, Cow
  • Immunogen

    Synthetic peptide within Human S6K1. The exact sequence is proprietary.

  • Positive control

    • Jurkat cell lysate; human breast carcinoma
  • 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

  • Form

    Liquid
  • Storage instructions

    Shipped at 4°C. Upon delivery aliquot and store at -20°C. Avoid freeze / thaw cycles.
  • Storage buffer

    pH: 7.20
    Preservative: 0.05% Sodium azide
    Constituents: 40% Glycerol (glycerin, glycerine), 9.85% Tris glycine, 50% Tissue culture supernatant
  • Concentration information loading...
  • Purity

    Protein A purified
  • Clonality

    Monoclonal
  • Clone number

    E175
  • Isotype

    IgG
  • Research areas

    • Cell Biology
    • Cell Cycle
    • Cell Cycle Inhibitors
    • Rb
    • Epigenetics and Nuclear Signaling
    • DNA / RNA
    • DNA Damage & Repair
    • DNA Damage Response
    • DNA Damage Recognition
    • Epigenetics and Nuclear Signaling
    • DNA / RNA
    • Translation
    • Ribosome
    • Signal Transduction
    • Protein Phosphorylation
    • Ser / Thr Kinases
    • S6K Family
    • Metabolism
    • Types of disease
    • Obesity

Associated products

  • Alternative Versions

    • Anti-S6K1 antibody [E175] - BSA and Azide free (ab239810)
  • Compatible Secondaries

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

    • Rabbit IgG, monoclonal [EPR25A] - Isotype Control (ab172730)
  • Positive Controls

    • Jurkat whole cell lysate (ab7899)
  • Recombinant Protein

    • Recombinant human S6K1 protein (ab62291)

Applications

The Abpromise guarantee

Our Abpromise guarantee covers the use of ab32359 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 - 1/5000. Detects a band of approximately 70 kDa (predicted molecular weight: 59 kDa).
IHC-P (2)
1/50. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.
Notes
WB
1/1000 - 1/5000. Detects a band of approximately 70 kDa (predicted molecular weight: 59 kDa).
IHC-P
1/50. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.
Application notes
Is unsuitable for Flow Cyt or ICC/IF.

Target

  • Function

    Acts to integrate nutrient and growth factor signals in regulation of protein synthesis, cell proliferation, cell growth, cell cycle progression and cell survival. Downstream effector of the mTOR signaling pathway. Phosphorylates specifically ribosomal protein S6 in response to insulin or several classes of mitogens. During translation initiation, the inactive form associatess with the eIF-3 complex under conditions of nutrient depletion. Mitogenic stimulation leads to phosphorylation and dissociation from the eIF-3 complex and the free activated form can phosphorylate other translational targets including EIF4B. Promotes protein synthesis by phosphorylating PDCD4 at 'Ser-67' and targeting it for degradation. Phosphorylates RICTOR leading to regulation of mammalian target of rapamycin complex 2 (mTORC2) signaling; probably phosphorylates RICTOR at 'Thr-1135'. Phosphorylates IRS1 at multiple serine residues coupled with insulin resistance; probably phosphorylates IRS1 at 'Ser-270'. Required for TNF-alpha induced IRS-1 degradation. Phosphorylates EEF2K in response to IGF1 and inhibits EEF2K activity. Phosphorylates BAD at 'Ser-99' in response to IGF1 leading to BAD inactivation and inhibition of BAD-induced apoptosis. Phosphorylates mitochondrial RMP leading to dissociation of a RMP:PPP1CC complex; probably phosphorylates RMP at 'Ser-99'. The free mitochondrial PPP1CC can dephosphorylate RPS6KB1 at Thr-412 which is proposed to be a negative feed back mechanism for the RPS6KB1 antiapoptotic function. Phosphorylates GSK3B at 'Ser-9' under conditions leading to loss of the TSC1-TSC2 complex. Phosphorylates POLDIP3.
  • Tissue specificity

    Widely expressed.
  • Sequence similarities

    Belongs to the protein kinase superfamily. AGC Ser/Thr protein kinase family. S6 kinase subfamily.
    Contains 1 AGC-kinase C-terminal domain.
    Contains 1 protein kinase domain.
  • Domain

    The autoinhibitory domain is believed to block phosphorylation within the AGC-kinase C-terminal domain and the activation loop.
    The TOS (TOR signaling) motif is essential for activation by mTORC1.
  • Post-translational
    modifications

    Phosphorylation at Thr-412 is regulated by mTORC1. The phosphorylation at this site is maintained by an agonist-dependent autophosphorylation mechanism.
  • Cellular localization

    Cytoplasm; Nucleus. Cytoplasm and Cell junction > synapse > synaptosome. Mitochondrion outer membrane.
  • Target information above from: UniProt accession P23443 The UniProt Consortium
    The Universal Protein Resource (UniProt) in 2010
    Nucleic Acids Res. 38:D142-D148 (2010) .

    Information by UniProt
  • Database links

    • Entrez Gene: 404181 Cow
    • Entrez Gene: 6198 Human
    • Entrez Gene: 72508 Mouse
    • Entrez Gene: 83840 Rat
    • Omim: 608938 Human
    • SwissProt: P23443 Human
    • SwissProt: Q8BSK8 Mouse
    • SwissProt: P67999 Rat
    • Unigene: 463642 Human
    • Unigene: 394280 Mouse
    • Unigene: 446624 Mouse
    • Unigene: 479484 Mouse
    • Unigene: 4042 Rat
    see all
  • Alternative names

    • 70 kDa ribosomal protein S6 kinase 1 antibody
    • KS6B1_HUMAN antibody
    • p70 alpha antibody
    • P70 beta 1 antibody
    • p70 ribosomal S6 kinase alpha antibody
    • p70 ribosomal S6 kinase beta 1 antibody
    • p70 S6 kinase alpha antibody
    • P70 S6 Kinase antibody
    • p70 S6 kinase, alpha 1 antibody
    • p70 S6 kinase, alpha 2 antibody
    • p70 S6K antibody
    • p70 S6K-alpha antibody
    • p70 S6KA antibody
    • p70(S6K) alpha antibody
    • p70(S6K)-alpha antibody
    • p70-alpha antibody
    • p70-S6K 1 antibody
    • p70-S6K antibody
    • P70S6K antibody
    • P70S6K1 antibody
    • p70S6Kb antibody
    • PS6K antibody
    • Ribosomal protein S6 kinase 70kDa polypeptide 1 antibody
    • Ribosomal protein S6 kinase beta 1 antibody
    • Ribosomal protein S6 kinase beta-1 antibody
    • Ribosomal protein S6 kinase I antibody
    • RPS6KB1 antibody
    • S6K antibody
    • S6K-beta-1 antibody
    • S6K1 antibody
    • Serine/threonine kinase 14 alpha antibody
    • Serine/threonine-protein kinase 14A antibody
    • STK14A antibody
    see all

Images

  • Western blot - Anti-S6K1 antibody [E175] (ab32359)
    Western blot - Anti-S6K1 antibody [E175] (ab32359)

    Lane 1: Wild-type HAP1 cell lysate (20 µg)
    Lane 2: S6K1 knockout HAP1 cell lysate (20 µg)
    Lane 3: MCF7 cell lysate (20 µg)
    Lane 4: HEK293 cell lysate (20 µg)
    Lanes 1 - 4: Merged signal (red and green). Green - ab32359 observed at 68 kDa. Red - loading control, ab8245, observed at 37 kDa.
    ab32359 was shown to recognize S6K1 when S6K1 knockout samples were used, along with additional cross-reactive bands. Wild-type and S6K1 knockout samples were subjected to SDS-PAGE. ab32359 and ab8245 (loading control to GAPDH) were diluted 1/1000 and 1/10000 respectively and incubated overnight at 4°C. Blots were developed with goat anti-rabbit IgG (H + L) and goat anti-mouse IgG (H + L) secondary antibodies at 1/10000 dilution for 1 hour at room temperature before imaging.

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-S6K1 antibody [E175] (ab32359)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-S6K1 antibody [E175] (ab32359)

    Paraffin-embedded human breast carcinoma
    ab32359 at 1/50 dilution

    Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.

  • Western blot - Anti-S6K1 antibody [E175] (ab32359)
    Western blot - Anti-S6K1 antibody [E175] (ab32359)
    Anti-S6K1 antibody [E175] (ab32359) + HEK-293T (Human epithelial cell line from embryonic kidney transformed with large T antigen) whole cell lysate

    Predicted band size: 59 kDa

  • Anti-S6K1 antibody [E175] (ab32359)
    Anti-S6K1 antibody [E175] (ab32359)

Protocols

  • Recommended WB protocol with ab32359

Click here to view the general protocols

Datasheets and documents

  • SDS download

  • Datasheet download

    Download

References (51)

Publishing research using ab32359? Please let us know so that we can cite the reference in this datasheet.

ab32359 has been referenced in 51 publications.

  • Li G  et al. Osteocytic Connexin43 Channels Regulate Bone-Muscle Crosstalk. Cells 10:N/A (2021). PubMed: 33530465
  • Li T  et al. RNF167 activates mTORC1 and promotes tumorigenesis by targeting CASTOR1 for ubiquitination and degradation. Nat Commun 12:1055 (2021). PubMed: 33594058
  • Zhang X  et al. FOXO3a regulates lipid accumulation and adipocyte inflammation in adipocytes through autophagy : Role of FOXO3a in obesity. Inflamm Res 70:591-603 (2021). PubMed: 33891136
  • Lin F  et al. Rapamycin protects against aristolochic acid nephropathy in mice by potentiating mammalian target of rapamycin-mediated autophagy. Mol Med Rep 24:N/A (2021). PubMed: 33955513
  • Thomas A  et al. Assessment of PI3K/mTOR/AKT Pathway Elements to Serve as Biomarkers and Therapeutic Targets in Penile Cancer. Cancers (Basel) 13:N/A (2021). PubMed: 34066040
View all Publications for this product

Customer reviews and Q&As

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

1-10 of 40 Abreviews or Q&A

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) abreview for Anti-S6K1 antibody [E175]

Average
Abreviews
Abreviews
Application
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Sample
Mouse Tissue sections (Liver)
Antigen retrieval step
Heat mediated - Buffer/Enzyme Used: CC2
Permeabilization
No
Specification
Liver
Blocking step
ventana optiview kit as blocking agent for 12 minute(s) · Concentration: 40µg/mL · Temperature: 37°C
Fixative
Formaldehyde
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 Feb 16 2021

Question

Ordered ab32359 not knowing the clone number had changed. Would like to switch out for ab129230

Read More

Abcam community

Verified customer

Asked on Nov 09 2012

Answer

Thank you for contacting Abcam.

I have gone ahead and refunded the cost of the order for ab32359 (see credit note information below) and also you should have received an email from FedEx with the shipping label for returning this antibody to Abcam.

Concerning the replacement antibody that we talked about, ab129230, as the original order was placed using a credit card and because our system does not store the credit card information, I cannot go ahead and place the order for the new antibody. If you could either call our CS team back with the credit card information, or place a new order through your purchasing agent for ab129230, we will get you that antibody as soon as possible.

I am sorry about this inconvenience and also for not spotting this issue whilst we talked on the phone, but I hope we can have this resolved fro you quickly.


Your credit note ID is **

If payment has already been made on the original order and you wish to receive a refund, please ask your purchasing department to contact our accounting department so that we may gather the correct information needed for the refund. To avoid confusion, please ensure your accounts department is aware of how the credit note is being used.

Our accounting department can be contacted by email at us.credits@abcam.com or by telephone using the information at the Contact Us link in the top right corner of our website. Please refer to the credit note ID in any correspondence with our accounting department.

The credit note ID is for your reference only and does not automatically guarantee the credit.

I hope this experience will not prevent you from purchasing other products from us in the future. Our Scientific Support team is always at your service, should you require further expert advice.

Read More

Abcam Scientific Support

Answered on Nov 09 2012

Question

I am looking for an antibody to S6K that reacts with drosophila.

Read More

Abcam community

Verified customer

Asked on Sep 26 2012

Answer

Thank you for contacting us. I can certainly compare the immunogens for our antibodies with a sequence for you. If you can provide the full length sequence you are interested in or the accession number, I will let you know which antibodies would be likely to cross-react.

Read More

Abcam Scientific Support

Answered on Sep 26 2012

Question

I am so sorry for the long delay. We have now finally desided which free og
charge antibody we like to try. Can you please send product# ab59208,
Anti-P70 S6 Kinase beta (phospho T229) antibody
Thanks in advance
Best regards

Read More

Abcam community

Verified customer

Asked on Sep 26 2012

Answer

Thank you for confirming these details and for your cooperation. The details provided enable us to closely monitor the quality of our products.
I am sorry this product did not perform as stated on the datasheet and for the inconvenience this has caused. As requested, I have issued a free of charge replacement with the order number 1171509 (ab59208).
To check the status of the order please contact our Customer Service team and reference this number.
Please note that this free of charge replacement vial is also covered by our Abpromise guarantee. Should you still be experiencing difficulties, or if you have any further questions, please do not hesitate to let us know.
I wish you the best of luck with your research.

Read More

Abcam Scientific Support

Answered on Sep 26 2012

Question

I am a Ph.D student studying at the University of Malta and I have some queries regarding my recent order:Anti-S6K antibody [E175] (ab32359) and Anti-AKT1 (phospho S473) antibody [EP2109Y] (ab81283).





To which S6K isoform (S6K1 and/or S6K2) does Anti-S6K antibody [E175] (ab32359) react to?





Moreover should the Anti-AKT1 (phospho S473) antibody [EP2109Y] (ab81283) stain normal tissue as I have been having the same intensities in tumour and in normal tissue. I have tried the Akt1 on cervix, breast and prostate tumours with surrounding normal tissue and in all cases it was difficult to determine between normal and tumour by the intensity.



I would like to thank you while awaiting for your reply

Read More

Abcam community

Verified customer

Asked on Sep 14 2012

Answer

Thank you for contacting us.


The exact epitope is proprietary, but the immunogen sequence is approximately 20 amino acids long and maps within amino acids 400-440 of p70 S6 Kinase of human origin.Thismeans, that both isoforms will be detected as they differ on the N-terminus.

I am attaching our questionnaire so that we can gather further information regarding the samples tested and the protocol used. Once we receive the completed questionnaire, we will look at the protocol and see if there are any suggestions we can make that may improve the results. This information will also allow us to investigate this case internally and initiate additional testing where necessary. If the product was purchased in the last six months and is being used according to our Abpromise, we would be happy to replace or refund the antibody.


I look forward to receiving your reply.

Read More

Abcam Scientific Support

Answered on Sep 14 2012

Question

wir haben einen Antikörper von Ihnen erhalten, der leider nicht funktioniert hat, weil es seine Phosphoprotein-Spezifität verloren hatte. Sie haben einen kostenlosen Ersatz oder eine Gutschrift angeboten. Wir möchten gerne den Gutschrift bekommen. Bitte senden Sie den an Frau Gerlinde Wimmelbacher von der Abteilung Einkauf.

Read More

Abcam community

Verified customer

Asked on Jul 31 2012

Answer

Vielen Dank für Ihre Antwort.

Die Referenznummer Ihrer Gutschrift ist 111111.

Es tut mir leid, dass der Antikörper nicht so funktioniert hat, wie auf dem Datenblatt angegeben. Ich habe unsere Finanzabteilung beauftragt, eine Gutschrift für Sie auszustellen. Sie können diese Gutschrift auf folgende Weise verwenden:

(1) Gegen die Rechnung dieses Antikörpers verwenden falls diese noch nicht bezahlt wurde
(2) Aufbewahren und gegen eine zukünftige Rechnung verwenden
(3) Wir stellen Ihnen gerne eine komplette Rückerstattung aus, wenn keine offenen Rechnungen mehr ausstehen

Um eine Rückerstattung des Kaufpreises zu erhalten oder um zu bestätigen wie Sie diese Gutschrift benutzen möchten, möchte ich Sie bitten, sich mit unserer Finanzabteilung in Verbindung zu setzen.
Sie können unsere Finanzabteilung per Email unter der Adresse creditcontrol@abcam.com oder telefonisch unter der Nummer 030 896 779 154 erreichen.

Bitte geben Sie die Referenznummer Ihrer Gutschrift immer mit an.

Die Referenznummer ist für Ihre Unterlagen bestimmt. Wir schicken Ihnen die eigentliche Gutschrift per Post oder Email zu. Sie enthält auch die Gutschriftnummer, die mit dem Buchstaben CGB beginnt.

Ich hoffe dass Sie trotz dieser schlechten Erfahrung weiterhin Kunde bei uns bleiben. Bitte zögern Sie nicht sich wieder an uns zu wenden, falls Sie weitere Fragen haben.

Read More

Abcam Scientific Support

Answered on Jul 31 2012

Question

Hi,
With regards to your email and previous discussions we wish to purchase the following antibodies free of charge
Anti-smad2 (ps467) #ab53100 £234
Anti-IKB (phosphor s32 + s36) ab 39a1431 #ab12135 £234
What is the best way to proceed with regards to ordering thse items? In addition, I know you have mentioned previously about another p-p70 s6 kinase we could use, could you possible provide me with this information again?
Thanks

Read More

Abcam community

Verified customer

Asked on Jun 27 2012

Answer

Thank you for getting back to me.
I can set up the order for the anti-smad2 (ps467) (ab53100) and anti-IKB (phospho s32 + s36) (ab12135) directly for you. Could you please just confirm that the delivery address I have for you is correct:
xxxxxxxx

Once this has been confirmed I will organise for the delivery.
The main alternative antibody we discussed directed against the phospho-T389 S6K was ab2571. Each new batch of this antibody is tested for its specificity by observing the change in band detected when a sample of HeLa cells are treated with and without l-phosphatase. As presented on the datasheet of this antibody, it has also been used with AEE788 treated rat samples indicating it shows strong phospho-specificity.
I hope this information has been of help. If you have any further questions, please do not hesitate to ask.
I look forward to receiving your reply.

Read More

Abcam Scientific Support

Answered on Jun 27 2012

Question

I would be interested in the ab2571 to replace the ab32359 antibody. My PO# was ***.



Thanks in advance

Read More

Abcam community

Verified customer

Asked on Jun 05 2012

Answer

Thank you for your reply.

I'm sending a free of charge vial of ab2571 on the order ***, which should ship out tonight and get to you tomorrow.

Please keep me updated about the performance of this replacement antibody, and let me know if there's anything else that we can do for you. Have a great day!

Read More

Abcam Scientific Support

Answered on Jun 05 2012

Question

Ab32359 was previously listed as being phospho-specific and that information has recently changed. Needs a phospho-specific S6K antibody to replace it.

Read More

Abcam community

Verified customer

Asked on Jun 05 2012

Answer

Thank you for your calls today and last week, and I am sorry for the trouble with our emails.

As we discussed, we have ab2571 and ab19380 which are both phospho-specific S6K antibodies. Please let me know which you would like as a replacement and I will be happy to send it out. Do you also have your order# or PO# that ab32359 waspurchased on?

I look forward to hearing from you. Please let me know if you have any questions or if there is anything else that we can do for you.

Read More

Abcam Scientific Support

Answered on Jun 05 2012

Question

die E-Mail des AK ab32359 vom 23.3.12 habe ich leider nicht erhalten.

E-Mails mit ungenauen Einkaufsdaten (Werbung etc.) landen oftmals im Spam, da nicht zu erkennen ist,

um welche Bestellung es sich handelt.

Bitte daher immer die Bestellnummer 305… od. 466… von uns mit angeben, damit ein Bezug erstellt werden kann.

Technische Probleme bitte immer mit dem Anwender absprechen.


Einkaufssachberarbeiterin

Read More

Abcam community

Verified customer

Asked on May 25 2012

Answer

Wenn wir eine Informationskampagne über einen Antikörper starten, versuchen wir natürlich immer, den Endverbraucher zu kontaktieren. Da aber viele Universitäten eine zentrale Einkaufsabteilung haben über die die Bestellungen abgewickelt werden, haben leider nicht die Kontaktdetails des Anwenders. Daher versenden wir die Information immer an die Email-Adresse, and die wir auch die Rechnung gesandt haben. Wir hoffen natürlich, dass dies dann entsprechend weitergeleitet wird, da es sehr wichtige Informationen sind.

Gerne würden wir Ihnen diesen "Papierkram" ersparen, wir denken aber, dass es wichtig ist, wenn unser Kunde über mögliche Komplikationen informiert worden ist. Des weiteren bieten wir bei Antikörpern, die große Qualitätsmängel aufweisen, immer einen kostenlosen Ersatz, eine Gutschrift oder eine Erstattung an, ganz egal, wann der Antikörper bestellt wurde.

Im Endeffekt bedeutet dies für den Endverbraucher, dass die Forschung ohne Probleme weitergeführt werden kann, ohne das weitere Arbeitsstunden und Material für eine Fehlerbehebung investiert werden müssen. (Und nebenher werden auch auch noch jede Menge Nerven geschont...)

Vielen Dank für Ihre Kooperation und Ihr Verständnis. Es ist uns bewusst, dass dieser nicht die optimale Lösung ist, aber glücklicherweise sind diese Maßnahmen nicht an der Tagesordnung.

PS: Wenn Sie keine Werbung/ Spam von uns erhalten möchten, kann ich Sie natürlich gern von unserer Werbeliste nehmen!

Read More

Abcam Scientific Support

Answered on May 25 2012

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