アブカムでは最適な動作のために Google Chrome など最新ブラウザでの閲覧を推奨します。

Hello. We're improving abcam.com and we'd welcome your feedback.

Hello. We're improving abcam.com and we'd welcome your feedback.

Infomation icon

We haven't added this to the BETA yet

New BETA website

New BETA website

Hello. We're improving abcam.com and we'd welcome your feedback.

Take a look at our BETA site and see what we’ve done so far.

Switch on our new BETA site

Now available

Search and browse selected products

  • A selection of primary antibodies

Purchase these through your usual distributor

In the coming months

  • Additional product types
  • Supporting content
  • Sign in to your account
  • Purchase online
お問い合わせは電話 +81-(0)3-6231-0940 または メールでどうぞ

  • アカウント
  • ログアウト
ログイン または 新規登録

Welcome

ログイン または

すでにアカウントをお持ちですか?

新規登録
カート
クイック・オーダー
Abcam homepage

  • 研究用製品
    製品タイプ別
    一次抗体
    二次抗体
    イムノアッセイキット・試薬
    細胞・組織イメージング
    細胞・バイオケミカルアッセイ
    タンパク質・ペプチド
    製品タイプ別
    プロテオミクス
    アゴニスト・アクチベータ-・アンタゴニスト・インヒビター
    細胞株・ライゼート
    miRNA マルチプレックス・アッセイキット
    マルチプレックス・アッセイ
    研究分野別
    癌
    循環器系
    細胞生物学
    エピジェネティクス
    メタボリズム
    発生生物学
    研究分野別
    免疫学
    微生物学
    脳神経科学
    シグナル伝達
    ステムセル
  • カスタム製品 & パートナーシップ
    カスタム製品 & パートナーシップ

    診断や治療プログラムを加速するための、カスタム製品および商業化に向けたパートナーシップ

    カスタム製品

    私たちとパートナーになる

  • サポート
    サポートハブ

    研究のためのサポートとアドバイス

    サポートハブを表示

    プロトコール

    実験のステップごとの詳細

    プロトコールを表示

  • イベント
    • イベント・カレンダー
    • 癌
    • 循環器
    • エピジェネティクス&核内情報
    • 免疫学
    • ニューロサイエンス
    • ステムセル
    • 展示会
    • ウェビナー
    最新のイベント情報

    世界中でアブカムが主催する研究会やセミナーの日程、内容、演者など

    イベント・カレンダー

  • パスウェイ
    細胞シグナル伝達パスウェイ

    すべてのパスウェイを見る

    インタラクティブ・パスウェイを見る

  1. Link

    mcm3-antibody-ab4460.pdf

  1. Send me a copy of this email
    I agree to the terms and conditions.
Epigenetics and Nuclear Signaling DNA / RNA DNA Synthesis Other
Share by email

Anti-MCM3 抗体 (ab4460)

  • Datasheet
  • SDS
Reviews (2)Q&A (2)References (23)

Product price, shipping and contact information

Currently unavailable

申し訳ございませんが、現在表示できません。

こちらにお問い合わせいただくか、またはしばらく経ってから改めてお試しください

 

Loading size & price…

 

出荷および受注について

Shipping info

Promotion Information

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-MCM3 antibody (ab4460)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-MCM3 antibody (ab4460)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-MCM3 antibody (ab4460)
  • Immunoprecipitation - Anti-MCM3 antibody (ab4460)
  • Western blot - Anti-MCM3 antibody (ab4460)
  • Western blot - Anti-MCM3 antibody (ab4460)
  • Immunocytochemistry/ Immunofluorescence - Anti-MCM3 antibody (ab4460)

Key features and details

  • Rabbit polyclonal to MCM3
  • Suitable for: ICC/IF, WB, IHC-P
  • Reacts with: Mouse, Human
  • Isotype: IgG

リコンビナント抗体で、ロット間での高い再現性を実現

Product image
Anti-MCM3 antibody [EPR7080] (ab128923)
  • 異なるロット間での安定した再現性
  • 容易なスケールアップ
  • 評価試験による特異性の確認済み
  • 倫理基準に準拠 - アニマル・フリーの生産

製品の概要

  • 製品名

    Anti-MCM3 antibody
    MCM3 一次抗体 製品一覧
  • 製品の詳細

    Rabbit polyclonal to MCM3
  • 由来種

    Rabbit
  • アプリケーション

    適用あり: ICC/IF, WB, IHC-Pmore details
  • 種交差性

    交差種: Mouse, Human
    交差が予測される動物種: Orangutan
  • 免疫原

    Synthetic peptide within Human MCM3 aa 500-600. The exact immunogen sequence used to generate this antibody is proprietary information. If additional detail on the immunogen is needed to determine the suitability of the antibody for your needs, please contact our Scientific Support team to discuss your requirements. NP_002379.2 (GeneID 4172).
    Database link: P25205

  • ポジティブ・コントロール

    • WB: HeLa cell RIPA extract. HeLa OHIO extract. IHC-P: Human colon carcinoma tissue. ICC: HeLa cells.
  • 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. Avoid freeze / thaw cycle.
  • バッファー

    pH: 7
    Preservative: 0.1% Sodium azide
    Constituents: 0.021% PBS, 1.764% Sodium citrate, 1.815% Tris
  • Concentration information loading...
  • 精製度

    Immunogen affinity purified
  • ポリ/モノ

    ポリクローナル
  • アイソタイプ

    IgG
  • 研究分野

    • Epigenetics and Nuclear Signaling
    • DNA / RNA
    • DNA Synthesis
    • Other

関連製品

  • 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)

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ab4460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
ICC/IF
Use a concentration of 1 µg/ml.
WB (2)
1/2000 - 1/10000. Detects a band of approximately 105 kDa (predicted molecular weight: 90 kDa).
IHC-P
1/1000 - 1/5000. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.
特記事項
ICC/IF
Use a concentration of 1 µg/ml.
WB
1/2000 - 1/10000. Detects a band of approximately 105 kDa (predicted molecular weight: 90 kDa).
IHC-P
1/1000 - 1/5000. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.

ターゲット情報

  • 機能

    Acts as component of the MCM2-7 complex (MCM complex) which is the putative replicative helicase essential for 'once per cell cycle' DNA replication initiation and elongation in eukaryotic cells. The active ATPase sites in the MCM2-7 ring are formed through the interaction surfaces of two neighboring subunits such that a critical structure of a conserved arginine finger motif is provided in trans relative to the ATP-binding site of the Walker A box of the adjacent subunit. The six ATPase active sites, however, are likely to contribute differentially to the complex helicase activity. Required for DNA replication and cell proliferation.
  • 配列類似性

    Belongs to the MCM family.
    Contains 1 MCM domain.
  • 翻訳後修飾

    Phosphorylated upon DNA damage, probably by ATM or ATR.
  • 細胞内局在

    Nucleus.
  • Target information above from: UniProt accession P25205 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: 4172 Human
    • Entrez Gene: 17215 Mouse
    • Omim: 602693 Human
    • SwissProt: P25205 Human
    • SwissProt: P25206 Mouse
    • Unigene: 179565 Human
    • Unigene: 4502 Mouse
    • 別名

      • Cervical cancer proto oncogene 5 antibody
      • DNA polymerase alpha holoenzyme associated P1 antibody
      • DNA polymerase alpha holoenzyme associated protein P1 antibody
      • DNA polymerase alpha holoenzyme-associated protein P1 antibody
      • DNA replication factor MCM3 antibody
      • DNA replication licensing factor mcm3 antibody
      • HCC 5 antibody
      • HCC5 antibody
      • hRlf beta subunit antibody
      • Human cervical cancer proto oncogene 5 antibody
      • MCM 3 antibody
      • mcm3 antibody
      • MCM3 minichromosome maintenance deficient 3 antibody
      • MCM3_HUMAN antibody
      • MGC1157 antibody
      • Minichromosome maintenance complex component 3 antibody
      • Minichromosome maintenance deficient 3 antibody
      • Minichromosome maintenance protein 3 antibody
      • P1 h antibody
      • P1 MCM3 antibody
      • P1 Protein antibody
      • P1-MCM3 antibody
      • P1.h antibody
      • p102 antibody
      • P102 protein antibody
      • Replication licensing factor beta subunit antibody
      • RLF beta subunit antibody
      • RLF subunit beta antibody
      • RLFB antibody
      see all

    画像

    • Western blot - Anti-MCM3 antibody (ab4460)
      Western blot - Anti-MCM3 antibody (ab4460)

      Whole cell lysate (10 µg) from HeLa, Jurkat, HEK-293T, and TCMK-1 cells prepared using NETN lysis buffer.

      ab4460 used for WB at 0.04 µg/ml.

    • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-MCM3 antibody (ab4460)
      Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-MCM3 antibody (ab4460)

      Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human small cell lung cancer tissue labelling MCM3 with ab4460 at 1/1000 (1µg/ml). Detection: DAB.

    • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-MCM3 antibody (ab4460)
      Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-MCM3 antibody (ab4460)
      Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human colon carcinoma tissue labelling MCM3 with ab4460 at 1/1000 (1µg/ml). Detection: DAB.
    • Immunoprecipitation - Anti-MCM3 antibody (ab4460)
      Immunoprecipitation - Anti-MCM3 antibody (ab4460)

      Samples: Whole cell lysate (1.0 mg per IP reaction; 20% of IP loaded) from HeLa cells prepared using NETN lysis buffer.

      Lanes 1 and 2: ab4460 used for IP at 6 µg per reaction, two lots tested.

      Lane 3 Control IgG.

      For blotting immunoprecipitated MCM3, ab4460 was used at 0.04 µg/ml.

    • Western blot - Anti-MCM3 antibody (ab4460)
      Western blot - Anti-MCM3 antibody (ab4460)
      HeLa cell RIPA extract (50 µg) incubated with ab4460 used at 0.2 µg/ml for 1 hour. HeLa cell RIPA extract (50 µg) incubated with ab4460 used at 0.2 µg/ml for 1 hour.
    • Western blot - Anti-MCM3 antibody (ab4460)
      Western blot - Anti-MCM3 antibody (ab4460)
      WB using ab4460. Western blot analysis of ab4460 on HeLa OHIO extract.
    • Immunocytochemistry/ Immunofluorescence - Anti-MCM3 antibody (ab4460)
      Immunocytochemistry/ Immunofluorescence - Anti-MCM3 antibody (ab4460)
      ICC/IF image of ab4460 stained HeLa cells. The cells were 4% formaldehyde fixed (10 min) and then incubated in 1%BSA / 10% normal goat serum / 0.3M glycine in 0.1% PBS-Tween for 1h to permeabilise the cells and block non-specific protein-protein interactions. The cells were then incubated with the antibody (ab4460, 1µg/ml) overnight at +4°C. The secondary antibody (green) was Alexa Fluor® 488 goat anti-rabbit IgG (H+L) used at a 1/1000 dilution for 1h. Alexa Fluor® 594 WGA was used to label plasma membranes (red) at a 1/200 dilution for 1h. DAPI was used to stain the cell nuclei (blue) at a concentration of 1.43µM.

    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

    • SDS download

    • Datasheet download

      Download

    参考文献 (23)

    ab4460 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。

    ab4460 は 23 報の論文で使用されています。

    • Drosopoulos WC  et al. TRF2 Mediates Replication Initiation within Human Telomeres to Prevent Telomere Dysfunction. Cell Rep 33:108379 (2020). PubMed: 33176153
    • Prikrylova T  et al. 5-hydroxymethylcytosine Marks Mammalian Origins Acting as a Barrier to Replication. Sci Rep 9:11065 (2019). PubMed: 31363131
    • Yang Q  et al. Minichromosome maintenance 3 promotes hepatocellular carcinoma radioresistance by activating the NF-?B pathway. J Exp Clin Cancer Res 38:263 (2019). PubMed: 31208444
    • Zhang Y  et al. Histone H3K27 methylation modulates the dynamics of FANCD2 on chromatin to facilitate NHEJ and genome stability. J Cell Sci 131:N/A (2018). PubMed: 29760279
    • Charrasse S  et al. Ensa controls S-phase length by modulating Treslin levels. Nat Commun 8:206 (2017). PubMed: 28785014
    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

    1-4 of 4 Abreviews or Q&A

    Western blot abreview for Anti-MCM3 antibody - ChIP Grade

    Excellent
    Abreviews
    Abreviews
    abreview image
    Application
    Western blot
    Sample
    Human Cell lysate - whole cell (HeLa)
    Loading amount
    50 µg
    Specification
    HeLa
    Gel Running Conditions
    Reduced Denaturing (12%)
    Blocking step
    Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C
    Read More

    Abcam user community

    Verified customer

    投稿 Nov 08 2011

    Western blot abreview for Anti-MCM3 antibody - ChIP Grade

    Good
    Abreviews
    Abreviews
    abreview image
    Application
    Western blot
    Sample
    Human Cell lysate - whole cell (HeLa)
    Loading amount
    25 µg
    Specification
    HeLa
    Gel Running Conditions
    Reduced Denaturing (12%)
    Blocking step
    Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: RT°C
    Read More

    Dr. Mirjam Eckert

    Verified customer

    投稿 Oct 30 2009

    Question

    a couple of month back I ordered one of your antibodies, anti-MCM3 antibody – ChIP Grade (ab4460). It was announced to be suitable for Western Blotting as well as immunoprecipitation. In Western Blotting it works fantastically, but for some reason I´ve got stuck with co-immunprecipitation. Could you please send me the protocol or some further information, how you tested the immunoprecipitation with this antibody?

    I´m trying to couple this antibody to beads and immunoprecipitate the MCM complex as well as putative interacting proteins. I have an MCM7-antibody as a positive control to check if I pulled down the whole complex.

    I´m using Dynabeads ® (invitrogen) cause when I used agarose beads MCM3 as well as MCM7 bound unspecifically to the beads. By Ponceau S staining I could see that the antibodies were coupled to the beads, as I could see the bands for the heavy and light chains of the antibodies, but I couldn´t detect the MCM7 in the pull-down of MCM3. So either my bait doesn´t bind to the antibody or the bait does bind and only the complex is not stable enough and MCM7 is not bound to MCM3 anymore. Could you maybe help with some recommendations or suggestions how to do the IP? Could it be that the antibody bind denatured protein but not the native one, as it works perfectly for Western Blotting?

    Read More

    Abcam community

    Verified customer

    Asked on Nov 01 2012

    Answer

    Thank you for contacting us.
    I'm sorry to hear that the antibody does not seem to work in IP.

    The protocol used to test the antibody in IP is as follows:

    Preparation of Cell Lysate used for Immunoprecipitation

    Reagents needed:

    PBS (Phosphate Buffered Saline, pH 7.2)


    NaCl 8.0 g
    KCl 0.2 g
    NaH2PO4 0.23 g
    Na2HPO4 0.12 g



    Adjust the volume to 1 liter with distilled water.

    Store at 4-25 C.

    IP Lysis Buffer


    NaCl 7.31 g
    1 M Tris, pH 8 25 ml
    0.5 M EDTA, pH 8 5 ml
    10% NP-40 25 ml
    Distilled Water 445 ml



    Store at 4 C.

    Protease Inhibitors
    Reconstitute with 1 ml of dH2O. Use 50 mcl per 5 ml of lysis buffer to harvest cells.

    Procedure:

    Allow cells to grow to approximately 80% confluence on 100mm polystyrene tissue culture plates. Approximately 8*10^6 cells (one plate at 80% confluence) are needed per IP reaction.
    Wash cells two times in cold PBS.
    Lyse cells to release soluble cellular proteins using 500 mcl of cold lysis buffer containing 1X protease inhibitors per 100 mm plate. Scrape cells from the plates, transfer to 1.5 ml micro-centrifuge tubes, and place on ice for 30 minutes to ensure efficient lysis.
    Centrifuge lysates at 10,000 x g; 5 minutes.
    Remove and pool the supernatant. Determine protein concentration.




    Immunoprecipitation Protocol

    Reagents Needed:

    Immunoprecipitation (IP) lysis buffer
    Protease Inhibitors
    Primary Antibodies
    Normal IgG, negative control
    Protein A Sepharose Beads
    Cell Lysate
    Sample Buffer

    IP lysis Buffer
    12.5 ml 1M NaCl (250mM)
    2.5 ml 1M Tris (50mM)
    500ul 0.5M EDTA (5mM)
    2.5ml 10% NP-40
    32 ml dH20

    Protein A Beads
    Resuspend 400 mg of Protein A beads in 10 ml of distilled H2O. Mix well to resuspend. Spin at 250 rpm for 5 minutes. Wash 3X in 10 ml IP Lysis buffer. Resuspend to 10 ml with IP lysis buffer for a 20% solution. Use 100 mcl per IP reaction.

    4X Sample Buffer
    Glycerol 4.0 g
    Tris Base 0.68 g
    Tris HCL 0.67 g
    LDS 0.80 g
    EDTA 6 mg
    Brilliant Blue G250 2.5 mg
    Phenol red 2.5 mg
    Adjust volume to 10 ml with ultra pure water.

    Store at 4 C.

    1X Sample Buffer
    4X sample buffer 150 mcl
    1M DTT 60 mcl
    Distilled water 390 mcl

    Make fresh for each use.

    Procedure:

    Prepare cell lysate according to protocol. Place 500 mcl of the prepared cell lysate (1-3 mg/ml) into a 1.5 ml micro-centrifuge tube.
    To this tube add 2 to 10 mcg of the primary antibody (If using neat sera or an IgG fraction such as Protein-A purified antibody, larger amounts are likely to be required. For best results, optimal amounts of antibody should be empirically defined.)
    To a negative control reaction, add an equivalent amount of normal rabbit IgG.
    Add 100 mcl of a 20% Protein A suspension to the mixture of antibody and cell lysate. Rotate the immunoprecipitation reactions (end-to-end) for 3 hours at room temperature or overnight at 4 C.
    Centrifuge (200 x g; 5 minutes) to pellet the complex.
    Remove the supernatant and add 500 mcl cold cell lysis buffer. Centrifuge (200 x g; 5 minutes).
    Repeat wash step 6 twice more. After each centrifugation remove as much of the supernatant as possible.
    After removing the supernatant from the third wash, add 40 mcl of freshly prepared 1X sample buffer to each tube and heat at 90 C for 5 minutes.
    Continue with electrophoresis and immunoblotting as described under western blotting protocol. Load 8 to 16 mcl (20 to 40% of the IP reaction) to a polyacrylamide gel.



    Note: For optimal results, complete reduction of the sample is required. We recommend the use of 0.1 M DTT in SDS-PAGE sample buffer and immediately heating samples, loading and running gels.

    Some additional advice and suggestions:

    The antibody has been tested without conjugation to beads, thus I would suggest trying it that way as well. If the conjugation does not work efficiently, then the IP will not work either.

    It is also important to include the isotype control (ab37415, https://www.abcam.com/Rabbit-IgG-ChIP-Grade-ab37415.html) as negative control as this would give you a better idea of the non-specific background.
    I'd suggest trying various antibody-to-lysate ratios (e.g. 1/100 to 1/1000) as the optimal ratio needs to be determined empirically and can vary from lysate type to lysate type and from antibody to antibody.
    Furthermore, in case that this might be a problem, for avoiding the heavy and light chain bands in the WB, I'd recommend using protein A/G-HRP (ab7456, ab7460) instead of the secondary antibody in the WB.

    I hope this information is helpful to you. Please do not hesitate to contact us if you need any more advice or information.

    Use our products? Submit an Abreview. Earn rewards!
    https://www.abcam.com/abreviews

    Read More

    Abcam Scientific Support

    Answered on Nov 01 2012

    Question

    I have a question about your MCM3 polyclonals - based on the possible sequence identity of the peptides that you use in your immunizations, do you expect any of your MCM3 antibodies to possibly recognize the Drosophila MCM3 protein? Thanking in advance,

    Read More

    Abcam community

    Verified customer

    Asked on Aug 08 2007

    Answer

    Thank you for your enquiry. There is a very low probability that ab3724 would react with Drosophila MCM3 and very low to zero probability that ab3725 or ab4460 would react too. I hope this information will be useful. Should you require any other assistance, please do not hesitate to contact me.

    Read More

    Abcam Scientific Support

    Answered on Aug 08 2007

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

    技術情報や製品情報をメールで受け取る 登録
    製品・研究分野別
    • 癌
    • 循環器
    • 細胞生物学
    • 発生
    • エピジェネティクス&核内情報
    • 免疫学
    • メタボリズム
    • 感染症
    • ニューロサイエンス
    • シグナル伝達
    • ステムセル
    製品・タイプ別
    • 一次抗体
    • 二次抗体
    • バイオケミカルズ
    • アイソタイプ・コントロール
    • マルチカラー・セレクター
    • キット
    • ローディング・コントロール
    • ライゼート
    • ペプチド
    • タンパク質
    • スライド
    • タグ抗体・細胞マーカー
    • その他試薬
    サービス&サポート
    • サポート
    • よくあるお問い合わせ
    • プロトコール&トラブルシュート
    • ご購入に関して
    • お問い合わせフォーム
    • トレーニング
    • ターゲット別に見る
    アブカムについて
    • 企業ウェブサイト
    • 投資家の皆様へ
    • 企業情報
    • 採用情報
    • アブカムについて
    イベント
    • 展示会
    • カンファレンス
    International websites
    • abcam.com
    • abcam.cn

    Join with us

    • LinkedIn
    • facebook
    • Twitter
    • YouTube
    • Terms of sale
    • Website terms of use
    • Cookie policy
    • Privacy policy
    • Legal
    © 1998-2022 Abcam plc. All rights reserved.