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Anti-ORC4L 抗体 (ab9641)

Submit a review Q&A (3)References (11)
Western blot - Anti-ORC4L antibody - ChIP Grade (ab9641)
  • Western blot - Anti-ORC4L antibody - ChIP Grade (ab9641)
  • Western blot - Anti-ORC4L antibody - ChIP Grade (ab9641)
  • Western blot - Anti-ORC4L antibody - ChIP Grade (ab9641)

Key features and details

  • Goat polyclonal to ORC4L
  • Suitable for: WB
  • Reacts with: Mouse, Rat, Human
  • Isotype: IgG

製品の概要

  • 製品名

    Anti-ORC4L antibody
    ORC4L 一次抗体 製品一覧

  • 製品の詳細

    Goat polyclonal to ORC4L

  • 由来種

    Goat

  • アプリケーション

    適用あり: WBmore details

  • 種交差性

    交差種: Mouse, Rat, Human
    交差が予測される動物種: Cow, Dog

  • 免疫原

    Synthetic peptide corresponding to Human ORC4L aa 400-500 (C terminal). (NP_001177810.1; NP_001177811.1)
    Database link: NP_002543.2

    Run BLAST with BLAST the sequence with ExPASy Run BLAST with BLAST the sequence with NCBI

  • 特記事項

    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

製品の特性

  • 製品の状態

    Liquid

  • 保存方法

    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.

  • バッファー

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

  • 精製度

    Immunogen affinity purified

  • 特記事項（精製）

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

  • ポリ/モノ

    ポリクローナル

  • アイソタイプ

    IgG

  • 研究分野

アプリケーション

The Abpromise guarantee

Abpromise保証は、 次のテスト済みアプリケーションにおけるab9641の使用に適用されます

アプリケーションノートには、推奨の開始希釈率がありますが、適切な希釈率につきましてはご検討ください。

アプリケーション Abreviews 特記事項
WB
Use a concentration of 0.5 - 2 µg/ml. Detects a band of approximately 50 kDa (predicted molecular weight: 50 kDa).

1 hour primary incubation is recommended for this product.
特記事項
WB
Use a concentration of 0.5 - 2 µg/ml. Detects a band of approximately 50 kDa (predicted molecular weight: 50 kDa).

1 hour primary incubation is recommended for this product.

ターゲット情報

  • 機能

    Component of the origin recognition complex (ORC) that binds origins of replication. DNA-binding is ATP-dependent, however specific DNA sequences that define origins of replication have not been identified so far. ORC is required to assemble the pre-replication complex necessary to initiate DNA replication.

  • 関連疾患

    Defects in ORC4 are the cause of Meier-Gorlin syndrome type 2 (MGORS2) [MIM:613800]. MGORS2 is a syndrome characterized by bilateral microtia, aplasia/hypoplasia of the patellae, and severe intrauterine and postnatal growth retardation with short stature and poor weight gain. Additional clinical findings include anomalies of cranial sutures, microcephaly, apparently low-set and simple ears, microstomia, full lips, highly arched or cleft palate, micrognathia, genitourinary tract anomalies, and various skeletal anomalies. While almost all cases have primordial dwarfism with substantial prenatal and postnatal growth retardation, not all cases have microcephaly, and microtia and absent/hypoplastic patella are absent in some. Despite the presence of microcephaly, intellect is usually normal.

  • 配列類似性

    Belongs to the ORC4 family.

  • 細胞内局在

    Nucleus.
  • Information by UniProt

  • 参照データベース

    see all

  • 別名

    • Origin recognition complex, subunit 4, S. cerevisiae, homolog of antibody
    • FLJ46668 antibody
    • HSORC4 antibody
    • ORC 4 antibody
    • ORC 4L antibody
    • ORC 4P antibody
    • ORC4 antibody
    • ORC4_HUMAN antibody
    • ORC4L antibody
    • ORC4L protein antibody
    • ORC4P antibody
    • Origin recognition complex subunit 4 (yeast homolog) like antibody
    • Origin recognition complex subunit 4 antibody
    • Origin recognition complex subunit 4 like (yeast) antibody
    • Origin recognition complex subunit 4 like antibody
    • origin recognition complex, subunit 4 homolog antibody
    • Origin recognition complex, subunit 4, S. cerevisiae, homolog-like antibody
    see all

画像

  • Western blot - Anti-ORC4L antibody - ChIP Grade (ab9641)
    Western blot - Anti-ORC4L antibody - ChIP Grade (ab9641)
    Anti-ORC4L antibody (ab9641) at 0.5 µg/ml + Mouse heart lysate at 35 µg

    Predicted band size: 50 kDa

  • Western blot - Anti-ORC4L antibody - ChIP Grade (ab9641)
    Western blot - Anti-ORC4L antibody - ChIP Grade (ab9641)
    Anti-ORC4L antibody (ab9641) at 0.5 µg/ml + Rat Heart lysate in RIPA buffer at 35 µg

    Developed using the ECL technique.

    Predicted band size: 50 kDa
    Observed band size: 50 kDa

  • Western blot - Anti-ORC4L antibody - ChIP Grade (ab9641)
    Western blot - Anti-ORC4L antibody - ChIP Grade (ab9641)
    ab9641 (2µg/ml) staining of Jurkat lysate (RIPA buffer, 1.4E5 cells per lane). Detected by western blot using chemiluminescence.
  • Western blot - Anti-ORC4L antibody - ChIP Grade (ab9641)
    Western blot - Anti-ORC4L antibody - ChIP Grade (ab9641)
    All lanes : Anti-ORC4L antibody (ab9641) at 1 µg/ml

    Lane 1 : Jurkat (Human T cell lymphoblast-like cell line) Whole Cell Lysate
    Lane 2 : HepG2 (Human hepatocellular liver carcinoma cell line) Whole Cell Lysate
    Lane 3 : HEK293 (Human embryonic kidney cell line) Whole Cell Lysate

    Lysates/proteins at 10 µg per lane.

    Secondary
    All lanes : Rabbit polyclonal to Goat IgG - H&L - Pre-Adsorbed (HRP) at 1/3000 dilution

    Developed using the ECL technique.

    Performed under reducing conditions.

    Predicted band size: 50 kDa
    Observed band size: 49 kDa why is the actual band size different from the predicted?
    Additional bands at: 38 kDa. We are unsure as to the identity of these extra bands.


    Exposure time: 20 minutes

参考文献 (11)

ab9641 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。

ab9641 は 11 報の論文で使用されています。

  • Hossain M  et al. Multiple, short protein binding motifs in ORC1 and CDC6 control the initiation of DNA replication. Mol Cell 81:1951-1969.e6 (2021). PubMed: 33761311
  • Cheung MH  et al. Human NOC3 is essential for DNA replication licensing in human cells. Cell Cycle 18:605-620 (2019). PubMed: 30741601
  • Hossain M & Stillman B Opposing roles for DNA replication initiator proteins ORC1 and CDC6 in control of Cyclin E gene transcription. Elife 5:N/A (2016). IP ; Human . PubMed: 27458800
  • Zhang R  et al. Targeting GLI by GANT61 involves mechanisms dependent on inhibition of both transcription and DNA licensing. Oncotarget 7:80190-80207 (2016). ICC/IF ; Human . PubMed: 27863397
  • Sherstyuk VV  et al. Mapping of Replication Origins in the X Inactivation Center of Vole Microtus levis Reveals Extended Replication Initiation Zone. PLoS One 10:e0128497 (2015). WB . PubMed: 26038842
View all Publications for this product

レビューと Q&A

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レビューを送る 質問を送る

1-3 of 3 Abreviews or Q&A

Question

BATCH NUMBER -- NOT SPECIFIED -- ORDER NUMBER 202754 DESCRIPTION OF THE PROBLEM no signal SAMPLE a lymphoblastoid cell line nuclear extract PRIMARY ANTIBODY AbCam 9641, 20ul in 2ml sample, 4C, overnight DETECTION METHOD Sybr Green ANTIBODY STORAGE CONDITIONS aliquoted and stored at -20C SAMPLE PREPARATION Crosslink, then wash with PBS twice, incubate at cell lysate buffer (5mM PIPES, 85mM KCl, 25ul NP40, 1:100 Protease Inhibitor cocktail) for 10 min on ice, spin down, and resuspend in nuclear lysate buffer (50 mM Tris-HCl, ph8, 10mM EDTA, 1% SDS, 1:100 protease inhibitor cocktail), then sonicate X-CHIP or N-CHIP X-ChIP CROSSLINKING 1% formaldehyde, at 37C for 15 min DNA FRAGMENTATION sonication, size is enriched between 500bp to 2Kb as checked on a 1% agarose gel IP STEP 10 to 15 million cells in 200 ul nuclear lysate buffer, then dilute with 9 volume of ChIP dilution buffer (Upstate), add 160 ul Salmone Sperm DNA/Protein A Agarose Slurry, 4C, 30 min, rotation, then spin, save the supernantant, add in 20ul of antibody, 4C, rotation overnight, then add 120ul beads, 1hr at 4C, and then wash the beads with low salt, high salt, LiCl, and TE (Upstate), then elute with 1% SDS in 0.1M NaHCO3 we also try to conjugate beads and antibody first DECROSSLINKING 20 ul 5M NaCl was added to the sample (final concentation 0.2M), 65C for 4 hr DNA PURIFICATION Phenol Chloroform extraction PELLET AFTER PRECIPITATION EtOH precipitation PRIMERS TESTED ON GENOMIC/INPUT DNA Yes, on genomic DNA with Sybr Green, no primer dimer NO-TEMPLATE CONTROL IN PCR Yes water control HOW MANY TIMES HAVE YOU TRIED THE APPLICATION? 2 DO YOU OBTAIN THE SAME RESULTS EVERY TIME? Yes WHAT STEPS HAVE YOU ALTERED? in the 2nd trial, we conjugate the beads with the antibody first

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Answer

Thank you for your enquiry. I am sorry to hear that you have been having difficulties with this antibody. This antibody was assigned "ChIP grade" status following its application in the following publication; Todorovic V et al. Human origins of DNA replication selected from a library of nascent DNA. Mol Cell 19:567-75 (2005). PubMed: 16109380 I have examined the protocol that they employed and the protocol that you are using is indeed very similar. As a control I would like to ask whether you have been incorporating a control antibody such as anti-histone H3. Also can you tell me the positive control loci that you have been using? Do you have a locus that you are expecting ORC4L to be recruited to? Is ORC4L constantly associated with this locus, or recruited in a stepwise process? I look forward to hearing from you.

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Question

We want to purchase this product. However, the lab is requesting lot#35325. Is that lot still available and would we request it?

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Answer

Thank you for your enquiry. Unfortunately, lot #35325 is no longer in stock. Please rest assured, however, that we guarantee that ab9641 will work as stated on the online datasheet or we will provide a replacement or refund. Please contact us again if you have any additional questions.

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Question

BATCH NUMBER 44011 ORDER NUMBER A39762 DESCRIPTION OF THE PROBLEM No signal when using Anti-ORC4L, not even for the positive control. However I can detect ORC2. SAMPLE HeLa nuclear extract PRIMARY ANTIBODY Anti-ORC4L, abcam, 1:500 dilution, 90' incubation, wash once with TS buffer (10 mM Tris pH 8.0, 200 mM NaCl) wash twice with TS-NP40 (0.05% NP40), wash once with TS SECONDARY ANTIBODY Anti-goat, IgG-HRP, Santa Cruz, 1:2000, 90' incubtaion at RT, wash as the primary antibody. DETECTION METHOD ECLPlus POSITIVE AND NEGATIVE CONTROLS USED Use HeLa nuclear extract as positive control (which give strong signal with Anti-ORC2 Ab). ANTIBODY STORAGE CONDITIONS -20 C SAMPLE PREPARATION Protease inhibitors from Sigma. Mix sample with 2x SDS Loading buffer, heat for 5'. ELECTROPHORESIS/GEL CONDITIONS Reducing, 8-16% TRANSFER AND BLOCKING CONDITIONS Transfer O/N, 0.120 A HOW MANY TIMES HAVE YOU TRIED THE APPLICATION? 3 HAVE YOU RUN A "NO PRIMARY" CONTROL? No DO YOU OBTAIN THE SAME RESULTS EVERY TIME? Yes WHAT STEPS HAVE YOU ALTERED? Used Anti-ORC4 at 1:250 dilution, avoid Sodium azide, incubate primary over night at 4 C, incubate secondary over night at 4 C, new ECLPlus kit.

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Answer

Thank you for your e-mail. I'm really sorry to hear you are having problems with this antibody and would like to give you a few suggestions. You have not mentioned the amount of protein loaded onto the gel. If it is under 30-40ug this may be too low and hence there is very little ORC4L protein to be detected. You may also have a very low concentration of ORC4L in your samples and a western blot detection may not be enough to detect the protein. I would recommend trying a cell line which is known to express ORC4L at detectable levels, such as Jurkat and NSO (mouse) cell lysates (we have Jurkat cell lysat in our catalogue:ab7899). Can I confirm also that you add Tween 20 in the buffer in which the primary antibody is diluted in, as it helps the penetration of the antibody into the membrane? Do you block the membrane with milk or BSA? Finally, our datasheet recommends trying 1-3ug/ml so I would suggest you try 3ug/ml and maybe 5ug/ml too? This may help detecting any low amounts of protein you may have in your samples. I hope this information helps, but please do not hesitate to contact me if you need further advice, Good luck with your westerns!

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