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Epigenetics and Nuclear Signaling Chromosome Structure Telomeres

Anti-TRF1抗体(ab1423)

  • Datasheet
  • SDS
Reviews (4)Q&A (12)References (38)

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Western blot - Anti-TRF1 antibody (ab1423)
  • Immunocytochemistry - Anti-TRF1 antibody (ab1423)
  • Immunocytochemistry - Anti-TRF1 antibody (ab1423)

Key features and details

  • Rabbit polyclonal to TRF1
  • Suitable for: ICC, WB
  • Reacts with: Human
  • Isotype: IgG

选择批间可重复性更高的重组抗体

Product image
Anti-TRF1 antibody [EPR6313] (ab129177)
  • 研究可靠 —— 各批次间结果一致且可重复
  • 长期批量供应 —— 采用重组技术,可实现快速生产
  • 首次实验即可成功 —— 经过大量验证确认了特异性
  • 符合伦理标准 —— 产品不含动物成分

概述

  • 产品名称

    Anti-TRF1抗体
    参阅全部 TRF1 一抗
  • 描述

    兔多克隆抗体to TRF1
  • 宿主

    Rabbit
  • 经测试应用

    适用于: ICC, WBmore details
    不适用于: IHC-P
  • 种属反应性

    与反应: Human
  • 免疫原

    Recombinant full length protein corresponding to Human TRF1. Human full length recombinant protein expressed in bacteria.

  • 常规说明

    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

性能

  • 形式

    Liquid
  • 存放说明

    Shipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C or -80°C. Avoid freeze / thaw cycle.
  • 存储溶液

    pH: 7.20
    Preservative: 0.05% Sodium azide
  • Concentration information loading...
  • 纯度

    Whole antiserum
  • 克隆

    多克隆
  • 同种型

    IgG
  • 研究领域

    • Epigenetics and Nuclear Signaling
    • Chromosome Structure
    • Telomeres
    • Tags & Cell Markers
    • Subcellular Markers
    • Nucleus
    • Chromatin

相关产品

  • ChIP Related Products

    • ChIP Kit (ab500)
  • Compatible Secondaries

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

    • Rabbit IgG, polyclonal - Isotype Control (ChIP Grade) (ab171870)
  • Recombinant Protein

    • Recombinant Human TRF1 protein (ab116406)

应用

The Abpromise guarantee

Abpromise™承诺保证使用ab1423于以下的经测试应用

“应用说明”部分 下显示的仅为推荐的起始稀释度;实际最佳的稀释度/浓度应由使用者检定。

应用 Ab评论 说明
ICC
Use at an assay dependent concentration.
WB (3)
1/2000. Detects a band of approximately 60-65 kDa. An additional non specific band at 35 kDa may be observed. Denatured whole cell extracts (prepare total cell extracts with 6M urea, 1% SDS, 150 mM NaCl, 25mM Tris ph8, and then sonicate for 10 seconds)and TBS based buffers with 0.5% Tween 20 are recommended.
说明
ICC
Use at an assay dependent concentration.
WB
1/2000. Detects a band of approximately 60-65 kDa. An additional non specific band at 35 kDa may be observed. Denatured whole cell extracts (prepare total cell extracts with 6M urea, 1% SDS, 150 mM NaCl, 25mM Tris ph8, and then sonicate for 10 seconds)and TBS based buffers with 0.5% Tween 20 are recommended.
应用说明
Is unsuitable for IHC-P.

靶标

  • 功能

    Binds the telomeric double-stranded TTAGGG repeat and negatively regulates telomere length. Involved in the regulation of the mitotic spindle. Component of the shelterin complex (telosome) that is involved in the regulation of telomere length and protection. Shelterin associates with arrays of double-stranded TTAGGG repeats added by telomerase and protects chromosome ends; without its protective activity, telomeres are no longer hidden from the DNA damage surveillance and chromosome ends are inappropriately processed by DNA repair pathways.
  • 组织特异性

    Highly expressed and ubiquitous. Isoform Pin2 predominates.
  • 序列相似性

    Contains 1 HTH myb-type DNA-binding domain.
  • 结构域

    The acidic N-terminal domain binds to the ankyrin repeats of TNKS1 and TNKS2. The C-terminal domain binds microtubules.
    The TRFH dimerization region mediates the interaction with TINF2.
  • 翻译后修饰

    Phosphorylated preferentially on Ser-219 in an ATM-dependent manner in response to ionizing DNA damage.
    ADP-ribosylation by TNKS1 or TNKS2 diminishes its ability to bind to telomeric DNA.
    Ubiquitinated by RLIM/RNF12, leading to its degradation by the proteasome. Ubiquitinated by a SCF (SKP1-CUL1-F-box protein) ubiquitin-protein ligase complex, leading to its degradation by the proteasome.
  • 细胞定位

    Nucleus. Cytoplasm > cytoskeleton > spindle. Chromosome > telomere. Colocalizes with telomeric DNA in interphase and metaphase cells and is located at chromosome ends during metaphase. Associates with the mitotic spindle.
  • Target information above from: UniProt accession P54274 The UniProt Consortium
    The Universal Protein Resource (UniProt) in 2010
    Nucleic Acids Res. 38:D142-D148 (2010) .

    Information by UniProt
  • 数据库链接

    • Entrez Gene: 7013 Human
    • Omim: 600951 Human
    • SwissProt: P54274 Human
    • Unigene: 442707 Human
    • 别名

      • hTRF1 AS antibody
      • NIMA interacting protein 2 antibody
      • NIMA-interacting protein 2 antibody
      • PIN 2 antibody
      • PIN2 antibody
      • t TRF1 antibody
      • Telomeric protein Pin2 antibody
      • Telomeric protein Pin2/TRF1 antibody
      • Telomeric repeat binding factor (NIMA interacting) 1 antibody
      • Telomeric repeat binding factor 1 antibody
      • Telomeric repeat binding protein 1 antibody
      • Telomeric repeat-binding factor 1 antibody
      • TERF 1 antibody
      • Terf1 antibody
      • TERF1_HUMAN antibody
      • TRBF 1 antibody
      • TRBF1 antibody
      • TRF 1 antibody
      • TRF antibody
      • TTAGGG repeat binding factor 1 antibody
      • TTAGGG repeat-binding factor 1 antibody
      see all

    图片

    • Western blot - Anti-TRF1 antibody (ab1423)
      Western blot - Anti-TRF1 antibody (ab1423)
      Western blot using anti-TRF1 at 1/2000. A single band at 60-65 kD is detected. Western blot using anti-TRF1 at 1/2000. A single band at 60-65 kD is detected.
    • Immunocytochemistry - Anti-TRF1 antibody (ab1423)
      Immunocytochemistry - Anti-TRF1 antibody (ab1423)

      Cells transfected with a TRF1-GFP contruct. On the left: immunofluorescence with the anti-TRF1 antibody used at 1/300, with a Texas Red-conjugated secondary antibody. In the centre: fluorescence from the GFP. Right: merge of both images showing overlap of GFP fluorescence and antibody staining.

    • Immunocytochemistry - Anti-TRF1 antibody (ab1423)
      Immunocytochemistry - Anti-TRF1 antibody (ab1423)
      The cells in the first image are probably in a stage of the cell cycle where the signal is not so clear. In this image the antibody is shown to recognise the same signal but staining is shown as 20-30 spots (punctate foci).

    实验方案

    • Immunocytochemistry & immunofluorescence protocols
    • Western blot protocols

    Click here to view the general protocols

    数据表及文件

    • SDS download

    • Datasheet download

      Download

    文献 (38)

    发表研究结果有使用 ab1423?请让我们知道,以便我们可以引用本数据表中的参考文章。

    ab1423 被引用在 38 文献中.

    • Ferreira da Silva J  et al. Prime editing efficiency and fidelity are enhanced in the absence of mismatch repair. Nat Commun 13:760 (2022). IF ; Human . PubMed: 35140211
    • Ge J  et al. Telomere Dysfunction in Oocytes and Embryos From Obese Mice. Front Cell Dev Biol 9:617225 (2021). IF ; Mouse . PubMed: 33553179
    • Kang S  et al. Transcriptional regulation of telomeric repeat-containing RNA by acridine derivatives. RNA Biol N/A:1-17 (2021). WB ; Human . PubMed: 33749516
    • Ali S  et al. Pt-ttpy, a G-quadruplex binding platinum complex, induces telomere dysfunction and G-rich regions DNA damage. Metallomics N/A:N/A (2021). ChIP ; Human . PubMed: 34021581
    • Morcos YAT  et al. A Novel Tissue and Stem Cell Specific TERF1 Splice Variant Is Downregulated in Tumour Cells. Int J Mol Sci 21:N/A (2019). WB, IF, IHC ; Mouse, Human . PubMed: 31877678
    View all Publications for this product

    客户评价及客户问答

    Show All 评价 Q&A
    提交评价 提交问题

    1-10 of 16 Abreviews or Q&A

    Western blot abreview for Anti-TRF1 antibody - ChIP Grade

    Poor
    Abreviews
    Abreviews
    abreview image
    Application
    Western blot
    Loading amount
    50 µg
    Gel Running Conditions
    Reduced Denaturing (8% resolution gel)
    Sample
    Human Cell lysate - whole cell (hepatocellular carcinoma)
    Specification
    hepatocellular carcinoma
    Treatment
    Inhibition of one gene's expression using lentiviral shRNA
    Blocking step
    Milk as blocking agent for 3 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 24°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

    提交于 Jul 16 2013

    Immunocytochemistry/ Immunofluorescence abreview for Anti-TRF1 antibody - ChIP Grade

    Average
    Abreviews
    Abreviews
    abreview image
    Application
    Immunocytochemistry/ Immunofluorescence
    Sample
    Human Cell (Breast carcinoma)
    Specification
    Breast carcinoma
    Fixative
    Paraformaldehyde
    Permeabilization
    Yes - 0.5% Triton X-100
    Blocking step
    Serum as blocking agent for 30 minute(s) · Concentration: 2% · Temperature: 37°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

    提交于 Aug 23 2009

    Western blot abreview for Anti-TRF1 antibody - ChIP Grade

    Poor
    Abreviews
    Abreviews
    abreview image
    Application
    Western blot
    Sample
    Human Cell lysate - whole cell (HeLa cell)
    Loading amount
    100 µg
    Specification
    HeLa cell
    Gel Running Conditions
    Reduced Denaturing
    Blocking step
    Milk as blocking agent for 30 minute(s) · Concentration: 5%
    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.

    Dr. Geun Hyoung Ha

    Verified customer

    提交于 May 27 2009

    Western blot abreview for Anti-TRF1 antibody - ChIP Grade

    Poor
    Abreviews
    Abreviews
    Application
    Western blot
    Sample
    Human Cell lysate - whole cell (Hela CELLS)
    Loading amount
    50 µg
    Specification
    Hela CELLS
    Gel Running Conditions
    Reduced Denaturing (8%)
    Blocking step
    Milk as blocking agent for 30 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.

    Mr. Geun-Hyoung Ha

    Verified customer

    提交于 Apr 30 2009

    Question

    Hi
    I’m surprise that this protocol is not the same as the one done by the technical sheet
    Indeed I did as recommended:
    - fixation with PFA 3.7% in PBS 5min

    - permeabilization with 0.5% triton-PBS for 5min

    - blocking with 5% milk/0.2% tween for 1h (I also try with 1% BSA in PBS for 30min)

    - incubation with antibody at 1/100 in 5% milk/0.2% tween (or in 0.1% BSA) for 1h at RT

    - incubation with secondary antibody for 30min at RT

    In a previous experiment I also try:

    - fixation with PFA 3.7% in PBS 10min

    - permeabilization with 0.2% triton-PBS for 10min

    - blocking with 1% BSA in PBS for 15min

    - incubation with antibody at 1/100 in PBS for 1h at RT

    - incubation with secondary antibody for 1h at RT


    And in all case, I didn’t obtain specific staining.

    Can I have a credit note/refund please?

    Read More

    Abcam community

    Verified customer

    Asked on Oct 17 2012

    Answer

    Thank you for your reply.

    Our datasheet for this antibody does not suggest any specific ICC protocol, as a general ICC protocol should work with this antibody. The protocol stated on the datasheet belongs to a customer’s review who submitted an Abreview for this antibody in that application, and that can actually serve as guidance for other researches.

    I understand, however, that this antibody should have given the expected results under the conditions you have used it, and it may well be that you have received a faulty vial.

    If the antibody is still under the 6 months guarantee I will be happy to either send you a replacement for the same or a different antibody, or, as you mentioned in your last email, refund the product’s price.

    Could you please send me your order number to be able to raise the replacement / refund?

    Thank you very much for your cooperation. I look forward to receiving your reply with further details.

    Read More

    Abcam Scientific Support

    回复于 Oct 17 2012

    Question

    The staining of TRF1 in MCF7 and another cell line is diffuse, and not primarily nuclear.

    Read More

    Abcam community

    Verified customer

    Asked on Jul 10 2012

    Answer

    Thank you for contacting us and bringing this to our attention. The switch to PFA fixation and detergent permeabilization instead of methanol fixation that I mentioned in our conversation may help, as that is what has worked for customers in the past.

    Please let us know if you continue to have difficulty.

    Read More

    Abcam Scientific Support

    回复于 Jul 10 2012

    Question

    I see a strong band at 50 kDa but only a very weak band at 60-65 kDa, where the antibody datasheet says this is observed. Other TRF1 antibody datasheets also show the protein at 60 kDa.

    Read More

    Abcam community

    Verified customer

    Asked on Jan 17 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 1016117.

    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

    回复于 Jan 17 2012

    Question

    follow up to previous correspondance: 1) Did the customer aliquot the vial upon receipt and uses a fresh aliquot every time? -> yes!! 2) Is the HeLa cell extract a nuclear extract? The protein is a nuclear protein and absence in the WB could be due to the low amount of TRF1 in the whole cell lysate. -> whole cell extract. but I tested TRF2 of other company, I detected TRF2 very well with whole cell extract. 3) Were the protease inhibitors added freshly to the lysis buffer? Were samples kept on ice at all times? -> of course! PIC was added and all samples kept on ice well. 4) Was the sample mixed with loading buffer and boiled? -> yes 5) You say "amount protein loaded: 50ug" is this for the WB only or for the IP too? -> No, I used 2mg for IP. Input was 50ug of protein. 6) Has the customer tried 5% BSA block? -> yes, but same result. 7) has the customer tried a higher dilution with the IPed samples? -> of course. I tried 1/500, 1/1000, 1/2000, 1/5000. 8)What is the buffer used to dilute the primary antibody and the secondary antibody? -> 5% milk(5% BSA also) and TBST only. 9) The WB image you provide says "TRF1 is not detected but orc4 is detected well". I see a faint band around the 55kDa mark which could be TRF1. The Swiss Prot database indicates that the MW of TRF1 is around 50kDa. The customer is happy with the faint band of orc4 but I would disagree and optimise the protocol for both antibodies. -> orc4 band was not faint. just scanner was bad.. and other our lab members see orc4 everyday. I think TRF1 antibody (delivered to me) has something wrong. 10) When investigating low molecular weight proteins the customer should use high percentage gels, use of a 7% gel means the bands are at the bottom of the gel and not well separated. -> but I detected well orc4(40kDa) and other proteins of low molecular (lower than TRF1) with 7% SDS gel. also, I tried again with 12% gel. but result was bad.. 11) for investigating the problem with the IP, I would need a detailed protocol of the IP. I suggest we first concentrate on the WB to find out if the antibody works. I believe the faint 50kDa band might be the TRF1 but an optimised protocol is necessary to get a clean band. The background from the IP samples could be due to the blocking agent used, the type of protein A or G beads used for the IP and other factors. Has the customer much experience of IP? -> 1 tube : whole cell extracts(1mg, 2mg of protein) were incubate with protein A beads (G beads also) for pre-clearing. 2 tube : lysis buffer and TRF1 antibody (diluted 1/500,1/1000, 1/2000, 1/5000.) were incubate with protein A beads (G beads also) for TRF1 antibody can attach to beads. ... after that, pre-cleared cell extract of 1 tube was transfered to protein A beads attached TRF1 antibody of 2 tube and incubated for 2,3,4,5hrs and O/N.

    Read More

    Abcam community

    Verified customer

    Asked on Jan 12 2005

    Answer

    Thank you for this information, it is very useful and I can now really understand the protocol and the optimisation steps. I think the WB protocol looks very good (apart from my suggestion of running a high percentage gel), the IP protocol we suggest is very different but the fact the customer has a problem in WB means the problem is not related to the IP protocol. I understand that the customer can detect TRF2 in whole cell extracts, but it is possible that the TRF1 levels are much lower and a nuclear extraction would concentrate the protein A/G-sepharose beads for 4 hours and finally boiling the sample . If you can provide me with the purchase order or Abcam order number I can send you a replacement antibody to try, but I would strongly recommend nuclear extraction to maximise the amount of TRF1 protein loaded on the gel. I would also suggest changes to the IP protocol: incubate the cell extracts with the antibody overnight (try 5ug antibody per 500ug lysate), then the next morning add the protein A sepharose beads, incubate together for 4hrs, then wash the beads and boil, separating the protein from the antibody and from the beads. Please do not hesitate to contact us for further details of our recommended IP protocol. I look forward to hearing from you,

    Read More

    Abcam Scientific Support

    回复于 Jan 12 2005

    Question

    Our end user claimed about ab1423. She performed western blot , and IP, but she coull not detect band. When she performed westernblot, she expected 60-65 KDa band according your data sheet. But she could detect 75KDa band. Please confirm this. And she performed IP, there was high back ground and no TRF1 band . I attached claim report and data. Check this.

    Read More

    Abcam community

    Verified customer

    Asked on Jan 05 2005

    Answer

    I have carefully looked at the protocol and unfortunately need more information to understand the problem, I am sorry for the delay. The antibody has not been tested on HeLa cells, it does cross react with human but it is known that cancer cells can be very different morphologically and biochemically, this may explain why the signal is low. 1) Did the customer aliquot the vial upon receipt and uses a fresh aliquot every time? 2) Is the HeLa cell extract a nuclear extract? The protein is a nuclear protein and absence in the WB could be due to the low amount of TRF1 in the whole cell lysate. 3) Were the protease inhibitors added freshly to the lysis buffer? Were samples kept on ice at all times? 4) Was the sample mixed with loading buffer and boiled? 5) You say "amount protein loaded: 50ug" is this for the WB only or for the IP too? 6) Has the customer tried 5% BSA block? 7) has the customer tried a higher dilution with the IPed samples? 8)What is the buffer used to dilute the primary antibody and the secondary antibody? 9) The WB image you provide says "TRF1 is not detected but orc4 is detected well". I see a faint band around the 55kDa mark which could be TRF1. The Swiss Prot database indicates that the MW of TRF1 is around 50kDa. The customer is happy with the faint band of orc4 but I would disagree and optimise the protocol for both antibodies. 10) When investigating low molecular weight proteins the customer should use high percentage gels, use of a 7% gel means the bands are at the bottom of the gel and not well separated. 11) for investigating the problem with the IP, I would need a detailed protocol of the IP. I suggest we first concentrate on the WB to find out if the antibody works. I believe the faint 50kDa band might be the TRF1 but an optimised protocol is necessary to get a clean band. The background from the IP samples could be due to the blocking agent used, the type of protein A or G beads used for the IP and other factors. Has the customer much experience of IP? I look forward to hearing from you, Thank you for your patience,

    Read More

    Abcam Scientific Support

    回复于 Jan 05 2005

    Question

    Hi, Thank you for your time and for all the information. My protocol is almost identical to this one that you gave me. I also tried two other protocols. If it looked like I was getting something close to what I expect then I would persevere but none of the changes that I have tried have made any difference - I just see the diffuse staining throughout the nucleus and cytoplasm. And so, I went to your website to go through the proceedings to return the product. However, when I clicked on the link to get the form in the 'return and replacement' policy under 'terms and conditions' the link does not take me to a form. Can you help me with this? Thank you,

    Read More

    Abcam community

    Verified customer

    Asked on Jul 13 2004

    Answer

    I'm sorry to hear that this antibody is not working out for you. If you send me the Abcam order number or purchase order number that was used for your order for ab1423, I can give you a refund. There is no need to return the product to us.

    Read More

    Abcam Scientific Support

    回复于 Jul 13 2004

    1-10 of 16 Abreviews or Q&A

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