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  1. Link

    irak-1-antibody-ab238.pdf

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Immunology Innate Immunity Cytokines Interleukins
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Anti-IRAK-1 抗体 (ab238)

  • Datasheet
Reviews (6)Q&A (15)References (16)

Product price, shipping and contact information

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出荷および受注について

Abpromise

保証された製品品質、優れたカスタマー・サポート。 詳細はこちら。

Western blot - Anti-IRAK-1 antibody (ab238)
  • Immunocytochemistry/ Immunofluorescence - Anti-IRAK-1 antibody (ab238)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-IRAK-1 antibody (ab238)
  • Immunocytochemistry/ Immunofluorescence - Anti-IRAK-1 antibody (ab238)
  • Immunocytochemistry/ Immunofluorescence - Anti-IRAK-1 antibody (ab238)

Key features and details

  • Rabbit polyclonal to IRAK-1
  • Suitable for: WB, IHC-P, ICC/IF
  • Reacts with: Mouse, Rat, Human, Rhesus monkey
  • Isotype: IgG

こちらの製品もご検討ください

二次抗体
Product image
Goat Anti-Rabbit IgG H&L (HRP) (ab205718)
標識
Product image
Alexa Fluor® 488 Conjugation Kit (Fast) - Lightning-Link® (ab236553)

関連製品

製品の概要

  • 製品名

    Anti-IRAK-1 antibody
    IRAK-1 一次抗体 製品一覧
  • 製品の詳細

    Rabbit polyclonal to IRAK-1
  • 由来種

    Rabbit
  • 特異性

    At least four isoforms of IRAK are known to exist; this antibody will detect all four isoforms. IRAK antibody is predicted to not cross-react with other members of the IRAK protein family.

  • アプリケーション

    適用あり: WB, IHC-P, ICC/IFmore details
  • 種交差性

    交差種: Mouse, Rat, Human, Rhesus monkey
  • 免疫原

    Synthetic peptide corresponding to Human IRAK-1 aa 700-712 (C terminal).
    (Peptide available as ab6230)

  • 特記事項

    Reproducibility is key to advancing scientific discovery and accelerating scientists’ next breakthrough.

    Abcam is leading the way with our range of recombinant antibodies, knockout-validated antibodies and knockout cell lines, all of which support improved reproducibility.

    We are also planning to innovate the way in which we present recommended applications and species on our product datasheets, so that only applications & species that have been tested in our own labs, our suppliers or by selected trusted collaborators are covered by our Abpromise™ guarantee.

    In preparation for this, we have started to update the applications & species that this product is Abpromise guaranteed for.

    We are also updating the applications & species that this product has been “predicted to work with,” however this information is not covered by our Abpromise guarantee.

    Applications & species from publications and Abreviews that have not been tested in our own labs or in those of our suppliers are not covered by the Abpromise guarantee.

    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, as well as customer reviews and Q&As.

製品の特性

  • 製品の状態

    Liquid
  • 保存方法

    Shipped at 4°C. Store at +4°C.
  • バッファー

    pH: 7.2
    Preservative: 0.02% Sodium azide
    Constituent: PBS
  • Concentration information loading...
  • 精製度

    Affinity purified
  • 特記事項(精製)

    Affinity chromatography purified via peptide column.
  • ポリ/モノ

    ポリクローナル
  • アイソタイプ

    IgG
  • 研究分野

    • Immunology
    • Innate Immunity
    • Cytokines
    • Interleukins
    • Signal Transduction
    • Protein Phosphorylation
    • Ser / Thr Kinases
    • Other Kinases
    • Cardiovascular
    • Atherosclerosis
    • Vascular Inflammation
    • Inflammatory mediators
    • Immunology
    • Innate Immunity
    • TLR Signaling

関連製品

  • Compatible Secondaries

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

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

    • Rabbit IgG, polyclonal - Isotype Control (ChIP Grade) (ab171870)
  • Positive Controls

    • THP1 whole cell lysate (ab7913)

アプリケーション

Our Abpromise guarantee covers the use of ab238 in the following tested applications.

The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.

アプリケーション Abreviews 特記事項
WB 1/1000. Predicted molecular weight: 77 kDa.

(THP-1 or HeLa whole cell) from non-activated cells.

IHC-P Use a concentration of 1 µg/ml. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.
ICC/IF Use a concentration of 10 µg/ml.

ターゲット情報

  • 機能

    Binds to the IL-1 type I receptor following IL-1 engagement, triggering intracellular signaling cascades leading to transcriptional up-regulation and mRNA stabilization. Isoform 1 binds rapidly but is then degraded allowing isoform 2 to mediate a slower, more sustained response to the cytokine. Isoform 2 is inactive suggesting that the kinase activity of this enzyme is not required for IL-1 signaling. Once phosphorylated, IRAK1 recruits the adapter protein PELI1.
  • 組織特異性

    Isoform 1 and isoform 2 are ubiquitously expressed in all tissues examined, with isoform 1 being more strongly expressed than isoform 2.
  • 配列類似性

    Belongs to the protein kinase superfamily. TKL Ser/Thr protein kinase family. Pelle subfamily.
    Contains 1 protein kinase domain.
  • 翻訳後修飾

    Autophosphorylated or is transphosphorylated by IRAK4 following recruitment to the IL-1RI. In the case of isoform 1, this is linked to ubiquitination and degradation.
    Polyubiquitinated; after cell stimulation with IL-1-beta. Polyubiquitination occurs with polyubiquitin chains linked through 'Lys-63'.
  • Target information above from: UniProt accession P51617 The UniProt Consortium
    The Universal Protein Resource (UniProt) in 2010
    Nucleic Acids Res. 38:D142-D148 (2010) .

    Information by UniProt
  • 参照データベース

    • Entrez Gene: 3654 Human
    • Entrez Gene: 16179 Mouse
    • Entrez Gene: 363520 Rat
    • Omim: 300283 Human
    • SwissProt: P51617 Human
    • SwissProt: Q62406 Mouse
    • Unigene: 522819 Human
    • Unigene: 38241 Mouse
    • Unigene: 22238 Rat
    see all
  • 別名

    • AA48924 antibody
    • Il1rak antibody
    • Interleukin 1 receptor associated kinase 1 antibody
    • Interleukin-1 receptor-associated kinase 1 antibody
    • IRAK antibody
    • IRAK-1 antibody
    • Irak1 antibody
    • IRAK1-S antibody
    • IRAK1_HUMAN antibody
    • mPLK antibody
    • OTTHUMP00000026014 antibody
    • OTTHUMP00000026015 antibody
    • OTTHUMP00000026020 antibody
    • OTTHUMP00000180621 antibody
    • Pelle antibody
    • Pelle homolog antibody
    • Pelle-like protein kinase antibody
    • Plpk antibody
    see all

画像

  • Western blot - Anti-IRAK-1 antibody (ab238)
    Western blot - Anti-IRAK-1 antibody (ab238)
    All lanes : Anti-IRAK-1 antibody (ab238) at 1 µg/ml

    Lane 1 : L1210 cell lysate
    Lane 2 : YB2/0 cell lysate

    Lysates/proteins at 15 µg per lane.

    Secondary
    All lanes : Goat anti-rabbit IgG HRP conjugate at 1/10000 dilution

    Predicted band size: 77 kDa
    Observed band size: 77 kDa



    1h incubation at RT in 5% NFDM/TBST.

  • Immunocytochemistry/ Immunofluorescence - Anti-IRAK-1 antibody (ab238)
    Immunocytochemistry/ Immunofluorescence - Anti-IRAK-1 antibody (ab238)
    ab238 at 10µg/ml staining Hela cells by ICC/IF
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-IRAK-1 antibody (ab238)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-IRAK-1 antibody (ab238)
    IHC image of ab238 staining in human breast carcinoma 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 ab238, 1µg/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.

  • Immunocytochemistry/ Immunofluorescence - Anti-IRAK-1 antibody (ab238)
    Immunocytochemistry/ Immunofluorescence - Anti-IRAK-1 antibody (ab238)Image courtesy of Dr Mahesh Shivananjappa by Abreview.

    ab238 staining IRAK-1 in murine RAW 264.7 cells by Immunocytochemistry/ Immunofluorescence.Cells were fixed in paraformaldehyde, permeabilized using 0.1% Triton-X100 in 2% BSA for 15 minutes, blocked with 2% BSA for 1 hour at 22°C and then incubated with ab238 at a 1/150 dilution for 16 hours at 4°C. The secondary used was an Alexa-Fluor 488 conjugated chicken anti-rabbit IgG (H+L) used at a 1/1000 dilution.

    See Abreview

  • Immunocytochemistry/ Immunofluorescence - Anti-IRAK-1 antibody (ab238)
    Immunocytochemistry/ Immunofluorescence - Anti-IRAK-1 antibody (ab238)

    Immunofluorescence of IRAK-1 in HeLa cells using ab238 at 20 ug/ml.

プロトコール

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

Click here to view the general protocols

データシートおよび資料

    • Datasheet
  • 参考文献 (16)

    ab238 を使用した論文を発表された方は、こちらまでお知らせください。データシートに掲載させていただきます。

    ab238 は 16 報の論文で使用されています。

    • Yang G & Zhao Y Overexpression of miR-146b-5p Ameliorates Neonatal Hypoxic Ischemic Encephalopathy by Inhibiting IRAK1/TRAF6/TAK1/NF-aB Signaling. Yonsei Med J 61:660-669 (2020). PubMed: 32734729
    • Chu H  et al. MicroRNA-206 promotes lipopolysaccharide-induced inflammation injury via regulation of IRAK1 in MRC-5 cells. Int Immunopharmacol 73:590-598 (2019). PubMed: 31279225
    • Ji L  et al. Jieduquyuziyin Prescription-Treated Rat Serum Suppresses Activation of Peritoneal Macrophages in MRL/Lpr Lupus Mice by Inhibiting IRAK1 Signaling Pathway. Evid Based Complement Alternat Med 2019:2357217 (2019). PubMed: 31781262
    • Lindeløv Vestergaard A  et al. MicroRNAs and histone deacetylase inhibition-mediated protection against inflammatory ß-cell damage. PLoS One 13:e0203713 (2018). PubMed: 30260972
    • Gao Y  et al. Overexpression of miR-146a blocks the effect of LPS on RANKL-induced osteoclast differentiation. Mol Med Rep 18:5481-5488 (2018). PubMed: 30387844
    View all Publications for this product

    レビューと Q&A

    Show All レビュー Q&A
    レビューを送る 質問を送る

    1-10 of 21 Abreviews or Q&A

    Immunocytochemistry/ Immunofluorescence abreview for Anti-IRAK antibody

    Excellent
    Abreviews
    Abreviews
    Application
    Immunocytochemistry/ Immunofluorescence
    Sample
    Rhesus monkey Cell (Colon)
    Specification
    Colon
    Fixative
    Paraformaldehyde
    Permeabilization
    Yes - 0.1% Triton-x-100
    Blocking step
    BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 3% · Temperature: 25°C
    Read More

    Abcam user community

    Verified customer

    投稿 Feb 21 2013

    Question

    Thank you a lot - I will let you know how it worked!!!

    Read More

    Abcam community

    Verified customer

    Asked on Dec 06 2012

    Answer

    I have checked your new order 1217270 and it looks that both antibodies (abhttp://ops.adminsite.com/admin/data/fs_ab.cfm?&intAbID=119289 and abhttp://ops.adminsite.com/admin/data/fs_ab.cfm?&intAbID=135027) are out of stock at the moment. We have reordered the items but it may take a week to get them. According to our record, we should be able to get them by 18th December.

    Could you please drop me an e-mail to confirm if you are happy to wait.

    I apologize for any inconvenience caused and look forward to hearing from you soon.

    Read More

    Abcam Scientific Support

    Answered on Dec 06 2012

    Question

    Thank you for the reply and helpful comments.

    I wouldd like to get the follwing products for replacement:
    Mouse monoclonal Anti-IRAK antibody [3F7] (ab119289)
    Rabbit polyclonal Anti-CARD10 antibody (ab135027)

    Our previous order is attaced as PDF, we did it on 10.08.12

    Thanks again for the help,

    Read More

    Abcam community

    Verified customer

    Asked on Dec 05 2012

    Answer

    Thank you for getting back to me.

    This is to confirm that I have placed a new order for you - for one vial of ab238 and another vial of ab64171 as free of charge replacements and the new order number is 1217226.

    I would kindly advise you to run a non-stimulated positive control cell line which expresses the target protein endogenously and to apply lysis buffer (i.e. RIPA) which contains a mixture of different proteinase inhibitors to prevent the degradation of the protein of interest.

    Recommended positive controls:

    ab238 - THP-1 or HeLa whole cell lysate;

    ab64171- Jurkat whole cell lysate

    I hope the second vials will perform better, and please do let me know how you are getting on with these two products.

    Good luck!

    Read More

    Abcam Scientific Support

    Answered on Dec 05 2012

    Question

    Sorry that siRNA file did not come through, I am sending it again. Please find siRNA experiment attached. Both CARD10 and IRAK1 are highly expressed in keratinocytes. There are downstream effects - knock-down of CARD10 results in reduction of IL8 and CCL5, knock-down of IRAK1 results in production of IL8. mRNA results are very nice.
    Please look also your own webpage: Your different CARD10 antibodies run on very different sizes - why it is like that? If you cannot really replace the antibodies, can we please get at least 50% reduction when we buy new ones? This is an important experiment thatt we need for publishing.

    Thank you for the help,

    Read More

    Abcam community

    Verified customer

    Asked on Dec 05 2012

    Answer

    Thank you for getting back to me and for passing some useful details. Your co-operation is much appreciated.

    As our Abpromise indicates, in the event that a product is not functioning in the applications/species cited on the product data sheet (and the problem has been reported within 6 months of purchase) we will happily offer a credit note/refund to the value of the product purchased.

    I can certainly offer replacement vials to you if you wish to carry on with the experiments.

    Could you please confirm the Abcam Order Number or you Purchase Order Number and the date of purchase.

    After reading through the detailed protocol you kindly forwarded to Abcam, I would like to make the following comments/suggestions:

    According to Swiss-Prot database, IRAK has 4 different isoforms (P51617-1, P51617-2, P51617-3, P51617-4) and these alternative splice variants have different sizes. You can find some further details at this site:

    http://www.uniprot.org/uniprot/P51617

    One more comment that RT-PCR data should be treated carefully since, there is no clear correlation between mRNA and protein levels and sometimes it is not easy to predict the protein expression levels from quantitative mRNA data.

    There are several publications available which delineate the technical boundaries of the approaches for quantitative analysis of protein expression and reveal that simple deduction from mRNA transcript analysis is insufficient.

    Please do let me know how you wish to proceed with your enquiry. I look forward to hearing from you soon.

    Read More

    Abcam Scientific Support

    Answered on Dec 05 2012

    Question

    Thank you for the reply. Here are my answers to your questions.

    About the siRNA experiment: siRNAs were purchased from Life Technologies, Transfected using lipofection (siPORT-NeoFX, Life Technologies agian) for 72 hours, some of the cells were induced with IFNgamma for 48 hours. For illustration, I add the results in Powerpoint file which shows that siRNAs work on RNA level and there are downstream effects on cytokine production. I also include WB picture again and our order as PDF.

    I do hope we can get replacement for these products with the following antibodies:

    Mouse monoclonal Anti-IRAK antibody [3F7] (ab119289)
    Rabbit polyclonal Anti-CARD10 antibody (ab135027)

    With best regards and thank you for the help.

    Read More

    Abcam community

    Verified customer

    Asked on Dec 04 2012

    Answer

    Thank you for getting back to me.

    I can find the 3 pdf files attached to your response but it seems that the ppt (Powerpoint) file did not come through.

    After reading through the detailed protocol you kindly forwarded to Abcam, I would like to make the following comments/suggestions:

    1) Control for siRNA experiments:

    Have you done a time-course experiment with siRNA treated cells to select the most optimal time point for the studies?
    How have you selected the sequence of siRNA? Have you tried more than one SiRNA sequence? Could you check if the sequence corresponds correctly to IRAK and CARD10 protein?
    Have you checked the transfection efficiency and optimized the length of time for knockout to take effect?
    Have you checked the transfection by fluorescence tags and the knockout by RT-PCR?
    Have you used the correct endogenous positive and negative controls? Have you applied scrambled siRNA or standard negative siRNA controls?



    2) RIPA buffer:

    Laemmli buffer does not contain any proteinase inhibitors to protect the target protein form degradation. I would suggest using RIPA buffer and you can find the composition of the this solution at this site:

    https://www.abcam.com/index.html?pageconfig=resource&rid=11379#A1



    3) Positive control:

    Since the cells are stimulated with IFN gamma, this may modify/alter the expression pattern of the target. It would be worth running good positive control alongside with the samples. It would be also be worth checking the cell viability after treatment.

    ab238 - THP-1 or HeLa whole cell lysate;

    ab64171- Jurkat whole cell lysate

    I hope this will be useful for you. Should you still have any problem with this antibody after following these suggestions, then please do not hesitate to contact our Technical Department again.

    Read More

    Abcam Scientific Support

    Answered on Dec 04 2012

    Question

    Hi,

    I am great fan of your antibodies. I have used them in many applications including ChIP, WB, IP, IF etc.
    But now we have got problem with two antibodies - the recognized bands are with wrong size. The result does not correspond with other results we have in mRNA level and according to downstream effects. Our other data is convincing and nice, but we cannot get the results with the antibodies listed here. The image is attached.
    1) CARD10 (ab64171) lot GR65252-4:
    116 kDa, observed band size can be : 130 kDa
    Our bands are ca 57 and ca 170
    2) IRAK (ab238) should be 80 kDa, ours is around 60 kDa, lot nr GR79890-1:
    Western:SDS-PAGE, sample in 1xSDS loading containing 100 mM DTT. Image is attaced.We have performed several experiments in different other conditions - no help!

    Can you please replace these products with other prodducts from your company?

    - Mouse monoclonal Anti-IRAK antibody [3F7] (ab119289)
    - Rabbit polyclonal Anti-CARD10 antibody (ab135027)


    With best regards and with hope we can solve the problem.

    Read More

    Abcam community

    Verified customer

    Asked on Dec 03 2012

    Answer

    I am sorry to hear that you have been experiencing problems using these two products (ab238 and ab64171) in the application that you wish. Thank you very much for sending the WB images.

    In order to assess the quality of our products I would ask that you complete a brief questionnaire relating to the application used. Often it is possible to make suggestions that may help resolve problems experienced using a particular product.

    As our Abpromise indicates, in the event that a product is not functioning in the applications/species cited on the product data sheet (and the problem has been reported within 6 months of purchase) we will happily offer a credit note/refund to the value of the product purchased.

    All our customer feedback, including complaints are monitored weekly by our in house technical support team. If a product is at fault the technical support team will consider removing the product from our catalogue in order to avoid future customer inconvenience.

    Could you please provide some further details of the protocol used and complete the following form (attached as a word document).

    I am particularly interested in the following information:

    - lysis buffer used,

    - positive control,

    - summary of the siRNA experiment (sequence for siRNA, transfection efficiency, endogenous positive and negative control)

    Thank you for your understanding and co-operation in this matter. I look forward to hearing from you soon and resolving this issue as soon as possible.

    Read More

    Abcam Scientific Support

    Answered on Dec 03 2012

    Question

    Hello:

    I purchased one antibody form ABCAM (AB238) for introcellular staining(for flow cyt). I saw the datasheet said the incubation time is 16 hours in 4 degree. In our routine protocol , this step for all antibodies is only 30 minutes in 4 degree, so why this one takes so long? is there some special binding character for this one?

    Thanks!

    Read More

    Abcam community

    Verified customer

    Asked on Nov 16 2012

    Answer

    Thank you for your inquiry.

    I can confirm that the protocol for flow cytometry that we know of for this antibody includes an incubation time of 16 hours at 4C.

    Please feel free to use your standard protocol if you find that this is working too for this antibody. In case the staining is week I can suggest too prolong the incubation time. A longer incubation will achieve an optimal saturation.

    If in your hand a shorter incubation time is working well, i would like to encourage you to also write an Abreview. Other researchers will value this information. And you would earn Abpoints that can be redeemed as discount on future purchases or as Amazon vouchers.

    I hope this information is helpful and wish you good luck with your research.

    Read More

    Abcam Scientific Support

    Answered on Nov 16 2012

    Question

    Thanks for the reply.
    I tried to altered the steps of blocking and primary antibody at RT or 4 degree and other way round, but results were the same.
    I purchased this antibody before 6 months, so I think could not get replace or refund.Thanks

    Read More

    Abcam community

    Verified customer

    Asked on Sep 12 2012

    Answer

    I am sorry to hear that the result did not improve.

    To proceed further in this case could you let me know, how the antibody was stored and what was the purchase order number?

    Thank very much for your cooperation in this case!

    Read More

    Abcam Scientific Support

    Answered on Sep 12 2012

    Question

    ReviewID: 31685 AbID: 238, Anti-IRAK antibody Rating: Inconclusive Image: Anti-IRAK antibody for Western blot (Mouse) Sample: Species: Mouse Type: Cell lysate - whole cell Loading amount: 50 µg Specification: RAW macrophages cell line Treatment: LPS and peptidoglycan Application: Application: Western blot Gel Running Conditions: Reduced Denaturing Blocking step: Milk as blocking agent for 1 hour at 25°C Blocking Concentration: 5% Other detail: Dilution: 1/500 Incubation time: 1 hour 30 minutes at 25°C Diluent: 5% milk in TBS-T Secondary Antibody: Name: Non-Abcam Antibody was used: sc-2030 Conjugation: HRP Dilution: 1/5000 Detection: Detection method: ecl+ Exposure: 5 minutes and 0 seconds Bands: Specific: no kDa Non-specific: ˜100KD kDa Additional Data:

    Read More

    Abcam community

    Verified customer

    Asked on Sep 11 2012

    Answer

    Thank you for submitting an Abreview of ab238. As you may have noticed, your review has now been published on our website.

    Since you obtained poor results using the antibody in a tested species and application, we would like to follow up on this to see if we can possibly improve the results you are seeing with this antibody.

    Based on the information in your review, I feel that altering the step in protocol may help toimprove your results. Did you try any troubleshooting of protocol?

    Additionally, this antibody is covered by our Abpromise, so if the antibody was purchased within the past 6 months, I would be happy to discuss a refund or replacement with you to resolve this issue.

    I look forward to your reply.

    Read More

    Abcam Scientific Support

    Answered on Sep 11 2012

    Western blot abreview for Anti-IRAK antibody

    Inconclusive
    Abreviews
    Abreviews
    abreview image
    Application
    Western blot
    Sample
    Mouse Cell lysate - whole cell (RAW macrophages cell line)
    Loading amount
    50 µg
    Specification
    RAW macrophages cell line
    Treatment
    LPS and peptidoglycan
    Gel Running Conditions
    Reduced Denaturing
    Blocking step
    Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C
    Read More

    Dr. Varkha Agrawal

    Verified customer

    投稿 Sep 11 2012

    1-10 of 21 Abreviews or Q&A

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