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Anti-PLK4抗体(ab2642)

  • Datasheet
  • SDS
Submit a review Q&A (7)References (10)

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Immunocytochemistry/ Immunofluorescence - Anti-PLK4 antibody (ab2642)
  • Western blot - Anti-PLK4 antibody (ab2642)
  • Flow Cytometry (Intracellular) - Anti-PLK4 antibody (ab2642)
  • Immunocytochemistry/ Immunofluorescence - Anti-PLK4 antibody (ab2642)

Key features and details

  • Goat polyclonal to PLK4
  • Suitable for: ICC/IF, WB, Flow Cyt (Intra)
  • Reacts with: Human
  • Isotype: IgG

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概述

  • 产品名称

    Anti-PLK4抗体
    参阅全部 PLK4 一抗
  • 描述

    山羊多克隆抗体to PLK4
  • 宿主

    Goat
  • 经测试应用

    适用于: ICC/IF, WB, Flow Cyt (Intra)more details
  • 种属反应性

    与反应: Human
    预测可用于: Mouse, Cow, Dog
  • 免疫原

    Synthetic peptide corresponding to Human PLK4 aa 1-100 (N terminal). (NP_001177728.1)
    Database link: O00444

    Run BLAST with BLAST the sequence with ExPASy Run BLAST with BLAST the sequence with NCBI
  • 阳性对照

    • Human colon lysate Flow Cyt (Intra): HeLa cells.
  • 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. Upon delivery aliquot and store at -20°C. Avoid freeze / thaw cycles.
  • 存储溶液

    pH: 7.30
    Preservative: 0.02% Sodium azide
    Constituents: Tris buffered saline, 0.5% BSA
  • Concentration information loading...
  • 纯度

    Immunogen affinity purified
  • 纯化说明

    Purified from goat serum by ammonium sulphate precipitation followed by antigen affinity chromatography using the immunizing peptide.
  • 克隆

    多克隆
  • 同种型

    IgG
  • 研究领域

    • Cell Biology
    • Cell Cycle
    • Cell Cycle Inhibitors
    • Other
    • Epigenetics and Nuclear Signaling
    • Cell cycle
    • Cell Cycle Inhibitors
    • Other

相关产品

  • Compatible Secondaries

    • Donkey Anti-Goat IgG H&L (Alexa Fluor® 488) (ab150129)
    • Donkey Anti-Goat IgG H&L (HRP) (ab205723)
  • Isotype control

    • Goat IgG, polyclonal - Isotype Control (ab37373)
  • Recombinant Protein

    • Recombinant human PLK4 protein (ab125558)

应用

The Abpromise guarantee

Abpromise™承诺保证使用ab2642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/IF
Use a concentration of 10 µg/ml.
WB
Use a concentration of 1 - 3 µg/ml. Detects a band of approximately 110 kDa (predicted molecular weight: 109 kDa).

1 hour primary incubation is recommended for this product.

Flow Cyt (Intra)
Use a concentration of 10 µg/ml.
说明
ICC/IF
Use a concentration of 10 µg/ml.
WB
Use a concentration of 1 - 3 µg/ml. Detects a band of approximately 110 kDa (predicted molecular weight: 109 kDa).

1 hour primary incubation is recommended for this product.

Flow Cyt (Intra)
Use a concentration of 10 µg/ml.

靶标

  • 功能

    Serine/threonine-protein kinase that plays a central role in centriole duplication. Able to trigger procentriole formation on the surface of the parental centriole cylinder, leading to the recruitment of centriole biogenesis proteins such as SASS6, CENPJ/CPAP, CP110, CEP135 and gamma-tubulin. When overexpressed, it is able to induce centrosome amplification through the simultaneous generation of multiple procentrioles adjoining each parental centriole during S phase. Its central role in centriole replication suggests a posible role in tumorigenesis, centrosome aberrations being frequently observed in tumors. Also involved in trophoblast differentiation by phosphorylating HAND1, leading to disrupt the interaction between HAND1 and MDFIC and activate HAND1. Phosphorylates CDC25C and CHEK2/CHK2.
  • 序列相似性

    Belongs to the protein kinase superfamily. Ser/Thr protein kinase family. CDC5/Polo subfamily.
    Contains 1 POLO box domain.
    Contains 1 protein kinase domain.
  • 翻译后修饰

    Ubiquitinated; leading to its degradation by the proteasome.
    Tyrosine-phosphorylated by TEC.
  • 细胞定位

    Cytoplasm > cytoskeleton > centrosome > centriole. Nucleus > nucleolus. Cleavage furrow. Associates with centrioles throughout the cell cycle. According to PubMed:16244668, it is not present at cleavage furrows.
  • Target information above from: UniProt accession O00444 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: 10733 Human
    • Entrez Gene: 20873 Mouse
    • SwissProt: O00444 Human
    • SwissProt: Q64702 Mouse
    • Unigene: 172052 Human
    • Unigene: 3794 Mouse
    • 别名

      • PLK 4 antibody
      • PLK-4 antibody
      • PLK4 antibody
      • PLK4_HUMAN antibody
      • Polo like Kinase 4 antibody
      • Polo-like kinase 4 antibody
      • Protein serine/threonine kinase antibody
      • SAK antibody
      • Serine/Threonine Kinase 18 antibody
      • Serine/threonine kinase antibody
      • Serine/threonine protein kinase 18 antibody
      • Serine/threonine protein kinase PLK4 antibody
      • Serine/threonine protein kinase Sak antibody
      • Serine/threonine-protein kinase 18 antibody
      • Serine/threonine-protein kinase PLK4 antibody
      • Serine/threonine-protein kinase SAK antibody
      • Snk Akin Kinase antibody
      • STK 18 antibody
      • STK18 antibody
      see all

    图片

    • Immunocytochemistry/ Immunofluorescence - Anti-PLK4 antibody (ab2642)
      Immunocytochemistry/ Immunofluorescence - Anti-PLK4 antibody (ab2642)
      Immunofluorescence analysis of paraformaldehyde fixed HeLa cells staining PLK4. Cells were permeabilized with 0.15% Triton. Samples were incubated with primary anitbody for 1 hour at 10µg/ml. An Alexa Fluor 488 was used as the secondary antibody. DAPI was used as a nuclear counterstain. Unimmunized goat IgG (10µg/ml) was used as the negative control.
    • Western blot - Anti-PLK4 antibody (ab2642)
      Western blot - Anti-PLK4 antibody (ab2642)
      Anti-PLK4 antibody (ab2642) at 2 µg/ml + human colon lysate in RIPA buffer at 35 µg

      Developed using the ECL technique.

      Predicted band size: 109 kDa
      Observed band size: 110 kDa why is the actual band size different from the predicted?


      Exposure time: 1 hour
    • Flow Cytometry (Intracellular) - Anti-PLK4 antibody (ab2642)
      Flow Cytometry (Intracellular) - Anti-PLK4 antibody (ab2642)
      Flow Cytometry analysis of HeLa cells labeling PLK4 with ab2642 at 10ug/mL followed by Alexa Fluor 488 secondary antibody (1ug/mL) (blue line). Cells were permeabilized with 0.5% Triton. IgG control: Unimmunized goat IgG (black line) followed by Alexa Fluor 488 secondary antibody.

       

    • Immunocytochemistry/ Immunofluorescence - Anti-PLK4 antibody (ab2642)
      Immunocytochemistry/ Immunofluorescence - Anti-PLK4 antibody (ab2642)
      Immunofluorescence analysis of paraformaldehyde fixed A431 cells staining PLK4. Cells were permeabilized with 0.15% Triton. Samples were incubated with primary anitbody for 1 hour at 10µg/ml. An Alexa Fluor 488 was used as the secondary antibody. DAPI was used as a nuclear counterstain. Unimmunized goat IgG (10µg/ml) was used as the negative control.

    实验方案

    • Recommended WB protocol for ab2642

    Click here to view the general protocols

    数据表及文件

    • SDS download

    • Datasheet download

      Download

    文献 (10)

    发表研究结果有使用 ab2642?请让我们知道,以便我们可以引用本数据表中的参考文章。

    ab2642 被引用在 10 文献中.

    • Yang Y  et al. CircKIF2A contributes to cell proliferation, migration, invasion and glycolysis in human neuroblastoma by regulating miR-129-5p/PLK4 axis. Mol Cell Biochem 476:2513-2525 (2021). PubMed: 33630225
    • Yang XD  et al. PLK4 deubiquitination by Spata2-CYLD suppresses NEK7-mediated NLRP3 inflammasome activation at the centrosome. EMBO J 39:e102201 (2020). PubMed: 31762063
    • Xu Z  et al. SNHG16 promotes tumorigenesis and cisplatin resistance by regulating miR-338-3p/PLK4 pathway in neuroblastoma cells. Cancer Cell Int 20:236 (2020). PubMed: 32536824
    • Shi J  et al. Long non-coding RNA RUNX1-IT1 plays a tumour-suppressive role in colorectal cancer by inhibiting cell proliferation and migration. Cell Biochem Funct 37:11-20 (2019). PubMed: 30499136
    • Zhang Z  et al. PLK4 is a determinant of temozolomide sensitivity through phosphorylation of IKBKE in glioblastoma. Cancer Lett 443:91-107 (2019). PubMed: 30529153
    • Czub B  et al. TOPORS, a Dual E3 Ubiquitin and Sumo1 Ligase, Interacts with 26 S Protease Regulatory Subunit 4, Encoded by the PSMC1 Gene. PLoS One 11:e0148678 (2016). PubMed: 26872363
    • Rosario CO  et al. A novel role for Plk4 in regulating cell spreading and motility. Oncogene N/A:N/A (2014). PubMed: 25174401
    • Rosario CO  et al. Plk4 is required for cytokinesis and maintenance of chromosomal stability. Proc Natl Acad Sci U S A 107:6888-93 (2010). PubMed: 20348415
    • Martindill DM  et al. Nucleolar release of Hand1 acts as a molecular switch to determine cell fate. Nat Cell Biol 9:1131-41 (2007). WB, ICC/IF ; Rat . PubMed: 17891141
    • Syed N  et al. Transcriptional silencing of Polo-like Kinase 2 (Snk/Plk2) is a frequent event in B cell malignancies. Blood : (2005). PubMed: 16160013

    客户评价及客户问答

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    1-7 of 7 Abreviews or Q&A

    Question

    Hi. If you say problems are few, I am willing to try a new batch of this antibody. Please fed-ex it to me. Thanks for your patience.

    Read More

    Abcam community

    Verified customer

    Asked on Nov 15 2004

    Answer

    I have organised a new vial to be sent to you, you should get a confirmation e-mail for this "order" soon. Please could you let me know how you get on with this batch? I would very much appreciate your feedback.

    Read More

    Abcam Scientific Support

    回复于 Nov 16 2004

    Question

    I will accept the new stock only if it has been recently tested and is known to work under your conditions. In perusing your website, I have noticed that I am not the only one with difficulties with this antibody. Does this antibody actually work? If you can state with confidence that this new batch is functional, I will accept this. Otherwise, I would like a credit note.

    Read More

    Abcam community

    Verified customer

    Asked on Nov 11 2004

    Answer

    The antibody in stock is a new batch and therefore has not been tested by other customers. I have arranged a credit note to be issued to you. This antibody sells well and the reported problems are therefore few. I wish you all the best with your research,

    Read More

    Abcam Scientific Support

    回复于 Nov 12 2004

    Question

    BATCH NUMBER 58237 ORDER NUMBER -- NOT SPECIFIED -- DESCRIPTION OF THE PROBLEM No signal at all SAMPLE 3T3 cells expressing recombinant flag-tagged human plk4 PRIMARY ANTIBODY 3 ug/mL anti plk-4, in 1% BSA tris-saline, 0.05% tween SECONDARY ANTIBODY 1:5000 anti-goat-hrp (santa cruz) DETECTION METHOD ECL POSITIVE AND NEGATIVE CONTROLS USED POSITIVE CONTROL: recombinant flag-tagged human plk4 was expressed in 3T3 cells and immunoprecipitated with anti-flag antibody. Membranes were probed with anti-flag antibody and a clear band of ~120 kDa was observed. When a parallel membrane was probed with anti-plk4, there is no band detected at all. NEGATIVE: 3T3 cells were transfected with empty vector (FLAG-CMV 7.0, sigma) and immunoprecipitated with anti-flag. When probed with anti-flag, there is no band at 120 kDa. ANTIBODY STORAGE CONDITIONS -20 SAMPLE PREPARATION Lysed at 4 degrees with protease and phosphatase inhibitors. 500 ug of protein immunoprecipitated with 2 ug of anti-flag antibody AMOUNT OF PROTEIN LOADED All of IP ELECTROPHORESIS/GEL CONDITIONS reducing 10% sds-page TRANSFER AND BLOCKING CONDITIONS semi-dry tris-glycine methanol HOW MANY TIMES HAVE YOU TRIED THE APPLICATION? 3 HAVE YOU RUN A "NO PRIMARY" CONTROL? Yes DO YOU OBTAIN THE SAME RESULTS EVERY TIME? Yes ADDITIONAL NOTES This antibody does not recognize plk-4 at all. I have tried this numerous times with whole cell lysate and recombinant proteins. I have wasted time and money and am very unhappy with tech support thus far. Please either send me a full refund or a new lot of the antibody that has actually been tested and is known to work.

    Read More

    Abcam community

    Verified customer

    Asked on Nov 10 2004

    Answer

    I'm terribly sorry you are having problems with ab2642. We have a new batch in stock which I could send to you or I can arrange a refund or credit note. Please let me know which option you would prefer and accept my apologies for this problem, I look forward to hearing from you,

    Read More

    Abcam Scientific Support

    回复于 Nov 11 2004

    Question

    I haven’t still get a real data following your suggested protocol. I have modified my Western bolt protocol as your described previously, including lyses placenta using RIPA buffer and no freeze-thawing to prevent protein degradation. But the result is the same as that the major band is still the wrong size about 66KDa when used HeLa cell lysate as sample. I also did the no primary control” which reveal that the major band 50KDa of human placenta lysate maybe come from the secondary antibody. I couldn’t still get my wanted band at appropriate 100KDa. I think this antibody didn’t work at all. You can test whether this antibody (lot #: 62683) is normal or not by yourself. Please offer me a replacement that is able to work normally. I'm looking forward to your next message.

    Read More

    Abcam community

    Verified customer

    Asked on Oct 18 2004

    Answer

    Thank you for your e-mail. I am very sorry you are not getting the right band, especially with your positive control. I will arrange for a new replacement vial to be sent to you. I understand you bought the antibody from Gene Research. Can you please confirm this and I will straight away arrange with them to send you a new vial. I look forward to hearing from you soon,

    Read More

    Abcam Scientific Support

    回复于 Oct 18 2004

    Question

    I am very happy to get your response. And following protocols were my detailed Western blotting analysis. ANTIBODY CODE ab2642 DESCRIPTION OF THE PROBLEM high background, the major band is the wrong size (66 KDa)?besides human placenta lysate that is at appropriate 50 KDa SAMPLE ?1?positive control as your datasheet using human placenta lysate? 2?various cancer cell lines including HeLa?PC3?HBL100?Au565?MCF7?A431 PRIMARY ANTIBODY incubated with anti-Plk4 antibody (1?2000) for overnight at 4? dissolving 0.3g BSA/10ml TBST buffer. Washed 4 times for 8 minutes using TBST buffer. SECONDARY ANTIBODY incubated with anti-goat conjugated-HRP antibody(1? 3000) for 40 minutes at room-temperature dissolving 0.3g milk/10ml TBST buffer. Washed 4 times for 6 minutes using TBST buffer. DETECTION METHOD used ECL detection system (Amersham) ANTIBODY STORAGE CONDITIONS Aliquot and store at -20? SAMPLE PREPARATION Cell lines and placenta tissue lysed with lysis buffer. And then freeze-thaw 3 times. Centrifuge at 15000 rpm for 10 minutes. The supernatant was subjected to the BCA Quantitation assay(BioRed).?lysis buffer?50mM Hepes-KOH pH7.4, 150mM NaCl, 1?TritonX-100, protease inhibitor ? AMOUNT OF PROTEIN LOADED 30 ug or 50 ug ELECTROPHORESIS/GEL CONDITIONS 7.5% SDS-PAGE gel TRANSFER AND BLOCKING CONDITIONS Transferred to PVDF membrane(MILLIPORE) in a trans-cassette. Blocked with 5?milk/TBST buffer at room-temperature for 1 hour. HOW MANY TIMES HAVE YOU TRIED THE APPLICATION? 5 HAVE YOU RUN A "NO PRIMARY" CONTROL? No, but I have used two different secondary antibodys from distinct sources. DO YOU OBTAIN THE SAME RESULTS EVERY TIME? Almost the same Above is my experimental protocol. Hope that you can find the root of the problem though these described information. Please help and contact me again as soon as possible. I'm looking forward to your next message.

    Read More

    Abcam community

    Verified customer

    Asked on Oct 08 2004

    Answer

    Thank you for the information. I have now a clearer idea of your problem and would like to help by suggesting some modifications to your protocol. I think the problem might be a degradation of your protein before it is recognised by the antibody as you mention freeze thawing it several times and I would like to recommend another lysis buffer (RIPA buffer = 50 mM Tris-HCl pH 7.4, 150 mM NaCl, 1 mM PMSF, 1 mM EDTA, 5 µg/ml Aprotinin, 5 µg/ml Leupeptin, 1% Triton -100, 1% Sodium deoxycholate, 0.1% SDS). Following homogenisation over 20 minutes on ice centrifuge at 13,000 rpm for 5 min in a centrifuge (which is pre-cooled and stays cool)transfer the supernatant into clean tubes and do the protein concentration as usual. Add an equal volume of 2 x SDS sample buffer (made with SDS and betamercaptoethanol) and boil for 5 minutes. You can then store the lysates at -80C until use. I think the rest of your WB protocol looks fine. To minimise the background you report I would suggest blocking your membrane in non-fat milk (3-5%w/v in TBST, filtered) for one hour and not incubating the other antibodies in blocking buffer (just TBST). After blocking, rinse once (for 10sec)the membrane with TBST.This will remove excess milk which can cause sometimes high background. It could also be due to too much secondary antibody or simply that the secondary has gone off...have you tried incubating with no primary, this will help determining if the problem comes from the secondary. Try also the secondary in other WBs for the proteins and see if it gives you problems. Also, wash extensively the membrane between steps, it's always better to wash many times for short periods of time (4-5min) than a few long washes. I hope this information helps, please do not hesitate to contact me again if you still have problems, Good luck!

    Read More

    Abcam Scientific Support

    回复于 Oct 08 2004

    Question

    Yes, we have done a "no primary" control and saw negilible binding to the membrane.How did you purify the protein for the test case? On the webpage you claimed it was human placental lysate, which I took to mean a whole cell extract.

    Read More

    Abcam community

    Verified customer

    Asked on Sep 10 2004

    Answer

    Thank you for your e-mail. Please find enclosed the protocol. Tissue Lysis: Tissue chunks were weighed and cut into approx 1mm cubes using a razor blade. The tissue was transferred to a handheld homogenizer and 3 ml of ice-cold RIPA buffer was added per 1g of tissue. The tissue was gently homogenised over 20 minutes on ice. The resulting lysate was aliquotted into 1.5 ml microfuge tubes and centrifuged at 13,000 rpm for 5 min in a microfuge. The supernatant was transferred into clean tubes and its protein concentration was measured with BioRad protein assay. The concentration was then adjusted to 5 mg/ml with RIPA lysis buffer. An equal volume of 2 x SDS sample buffer was added and the cell lysate was boiled for 5 minutes. Lysates were stored at -80C until use.(RIPA buffer = 50 mM Tris-HCl pH 7.4, 150 mM NaCl, 1 mM PMSF, 1 mM EDTA, 5 µg/ml Aprotinin, 5 µg/ml Leupeptin, 1% Triton -100, 1% Sodium deoxycholate, 0.1% SDS). We hope this will help.

    Read More

    Abcam Scientific Support

    回复于 Sep 16 2004

    Question

    ANTIBODY CODE ab2642 DESCRIPTION OF THE PROBLEM High background, major band is the wrong size (72 kDa) SAMPLE Multiple samples used: 1) Cell line BJ- human fibroblast line using exponentially growing cells and nocodazole treated cells 2) Human colorectal cancer tissue lysates 3) Human colorectal cancer cell lines PRIMARY ANTIBODY Incubated with 3ug/mL of your anti-plk4 antibody overnight at 4 degrees celcius in 1%BSA Tris/Saline + 0.05% Tween. Washed 3x for 5 minutes each with Tris/Saline with 0.05% Tween. SECONDARY ANTIBODY Used 1:2000 santa-cruz donkey anti-goat HRP conjugated antibody with 1%BSA Tris/Saline +0.05% Tween. Washed 3X for 5 minutes each with Tris/Saline with 0.05% Tween. DETECTION METHOD used amersham chemiluminescent detection system, femto grade POSITIVE AND NEGATIVE CONTROLS USED Positve control: M phase enriched cells (blocked 48 hrs with nocodazole) Negative control: G0 phase enriched cells (Blocked 48 hrs with serum starvation) ANTIBODY STORAGE CONDITIONS Aliquoted and stored at -20 SAMPLE PREPARATION Cell lines and tissues lysed in TNT with complete protease inhibitors (Roche) plus phosphatase inhibitors. Diluted with SDS-PAGE loading dye with 2-mercaptoethanol. Boiled at 100 degrees celcius for 10 minutes. AMOUNT OF PROTEIN LOADED 30 ug. ELECTROPHORESIS/GEL CONDITIONS Reducing 8% SDS-PAGE gel TRANSFER AND BLOCKING CONDITIONS Transferred to immbolion-p PDVF membranes using tris/glycine/methanol buffer in a mini-trans blot (BioRad) system. Blocked by drying in fume hood overnight. HOW MANY TIMES HAVE YOU TRIED THE APPLICATION? 4 HAVE YOU RUN A "NO PRIMARY" CONTROL? No DO YOU OBTAIN THE SAME RESULTS EVERY TIME? No ADDITIONAL NOTES I just can't seem to get a major band that is greater than 100 kDa... and my blots are very dirty. Please help!

    Read More

    Abcam community

    Verified customer

    Asked on Sep 04 2004

    Answer

    Thank you for your enquiry and for forwarding your detailed protocol. This antibody has been tested for Western blot application using Human Placenta Lysate. We would strongly suggest running a positive control along with the samples. Also you need to run "no primary" control to see if the secondary antibody gives a clear background. You have mentioned in your e-mail that "Blocked by drying in fume hood overnight" - we are very sorry but haven't heard of anyone doing this before. You may wish to try the following: after protein transfer on PVDF membrane, block the membrane in 2.5% skimmed milk in TBS-T for 1 hr at room temperature with agitation. Note that if PVDF membranes dry out after transfer therefore they should be "rewetted" with 100% methanol – there should be some information about this in the membrane product literature. We hope this will help. Should you still need more technical support, please do not hesitate to contact us again.

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    Abcam Scientific Support

    回复于 Sep 08 2004

    Please note: All products are "FOR RESEARCH USE ONLY. NOT FOR USE IN DIAGNOSTIC PROCEDURES"
    For licensing inquiries, please contact partnerships@abcam.com

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    研究类别(A-Z)
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