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遺伝子ノックアウト細胞株 検証済リコンビナントRabMAb

Anti-KMT5A / SETD8 / Pr-SET7 抗体 [EPR24381-24] - BSA and Azide free (ab300518)

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Western blot - Anti-KMT5A / SETD8 / Pr-SET7 antibody [EPR24381-24] - BSA and Azide free (AB300518)
  • Western blot - Anti-KMT5A / SETD8 / Pr-SET7 antibody [EPR24381-24] - BSA and Azide free (AB300518)
  • Immunoprecipitation - Anti-KMT5A / SETD8 / Pr-SET7 antibody [EPR24381-24] - BSA and Azide free (AB300518)
  • Flow Cytometry (Intracellular) - Anti-KMT5A / SETD8 / Pr-SET7 antibody [EPR24381-24] - BSA and Azide free (AB300518)
  • Anti-KMT5A / SETD8 / Pr-SET7 antibody [EPR24381-24] - BSA and Azide free (AB300518)

Key features and details

  • Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
  • Rabbit monoclonal [EPR24381-24] to KMT5A / SETD8 / Pr-SET7 - BSA and Azide free
  • Suitable for: IP, WB, Flow Cyt (Intra)
  • Knockout validated
  • Reacts with: Human

Conjugates logo Related conjugates and formulations

Unconjugated

製品の概要

  • 製品名

    Anti-KMT5A / SETD8 / Pr-SET7 antibody [EPR24381-24] - BSA and Azide free
    KMT5A / SETD8 / Pr-SET7 一次抗体 製品一覧

  • 製品の詳細

    Rabbit monoclonal [EPR24381-24] to KMT5A / SETD8 / Pr-SET7 - BSA and Azide free

  • 由来種

    Rabbit

  • アプリケーション

    適用あり: IP, WB, Flow Cyt (Intra)more details
    適用なし: ChIP,ICC/IF or IHC-P

  • 種交差性

    交差種: Human
    非交差種: Mouse, Rat

  • 免疫原

    Recombinant fragment. This information is proprietary to Abcam and/or its suppliers.

  • ポジティブ･コントロール

    • WB: HCT116, SH-SY5Y, U-2 OS, Wild-type HEK-293 and KMT5A knockout HEK-293 whole lysates. Flow Cyt (Intra): U-2 OS cell. IP: U-2 OS cell.

  • 特記事項

    ab300518 is the carrier-free version of ab300517.

     

    Our carrier-free antibodies are typically supplied in a PBS-only formulation, purified and free of BSA, sodium azide and glycerol. The carrier-free buffer and high concentration allow for increased conjugation efficiency.

    This conjugation-ready format is designed for use with fluorochromes, metal isotopes, oligonucleotides, and enzymes, which makes them ideal for antibody labelling, functional and cell-based assays, flow-based assays (e.g. mass cytometry) and Multiplex Imaging applications.

    Use our conjugation kits for antibody conjugates that are ready-to-use in as little as 20 minutes with <1 minute hands-on-time and 100% antibody recovery: available for fluorescent dyes, HRP, biotin and gold.

    This product is compatible with the Maxpar® Antibody Labeling Kit from Fluidigm, without the need for antibody preparation. Maxpar® is a trademark of Fluidigm Canada Inc.

    This product is a recombinant monoclonal antibody, which offers several advantages including:

    • - High batch-to-batch consistency and reproducibility
    • - Improved sensitivity and specificity
    • - Long-term security of supply
    • - Animal-free production
    For more information see here.

    Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMAb® patents.

製品の特性

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ab300518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
IP
Use at an assay dependent concentration.
WB
Use at an assay dependent concentration.
Flow Cyt (Intra)
Use at an assay dependent concentration.
特記事項
IP
Use at an assay dependent concentration.
WB
Use at an assay dependent concentration.
Flow Cyt (Intra)
Use at an assay dependent concentration.
追加情報
Is unsuitable for ChIP,ICC/IF or IHC-P.

ターゲット情報

  • 機能

    Protein-lysine N-methyltransferase that monomethylates both histones and non-histone proteins. Specifically monomethylates 'Lys-20' of histone H4 (H4K20me1). H4K20me1 is enriched during mitosis and represents a specific tag for epigenetic transcriptional repression. Mainly functions in euchromatin regions, thereby playing a central role in the silencing of euchromatic genes. Required for cell proliferation, probably by contributing to the maintenance of proper higher order structure of DNA during mitosis. Involved in chromosome condensation and proper cytokinesis. Nucleosomes are preferred as substrate compared to free histones. Mediates monomethylation of p53/TP53 at 'Lys-382', leading to repress p53/TP53-target genes.

  • 配列類似性

    Belongs to the histone-lysine methyltransferase family. PR/SET subfamily.
    Contains 1 SET domain.

  • 発生段階

    Not detected during G1 phase. First detected during S through G2 phases, and peaks during mitosis (at protein level).

  • ドメイン

    Although the SET domain contains the active site of enzymatic activity, both sequences upstream and downstream of the SET domain are required for methyltransferase activity.

  • 細胞内局在

    Nucleus. Chromosome. Specifically localizes to mitotic chromosomes. Associates with silent chromatin on euchromatic arms. Not associated with constitutive heterochromatin.
  • Information by UniProt

  • 参照データベース

    • 別名

      • H4 K20 HMTase antibody
      • H4 K20 specific histone methyltransferase antibody
      • H4-K20-HMTase SETD8 antibody
      • Histone H4 K20 methyltransferase antibody
      • Histone lysine N methyltransferase H4 lysine 20 specific antibody
      • Histone-lysine N-methyltransferase SETD8 antibody
      • Lysine N-methyltransferase 5A antibody
      • N-lysine methyltransferase SETD8 antibody
      • Pr SET 7 antibody
      • Pr SET7 antibody
      • PR-Set7 antibody
      • PR/SET domain containing protein 07 antibody
      • PR/SET domain containing protein 8 antibody
      • PR/SET domain-containing protein 07 antibody
      • PR/SET07 antibody
      • PrSET7 antibody
      • SET 07 antibody
      • SET 8 antibody
      • SET domain containing (lysine methyltransferase) 8 antibody
      • SET domain containing 8 antibody
      • SET domain containing lysine methyltransferase 8 antibody
      • SET domain containing protein 8 antibody
      • SET domain-containing protein 8 antibody
      • SET07 antibody
      • SET8 antibody
      • SETD 8 antibody
      • setd8 antibody
      • SETD8_HUMAN antibody
      see all

    画像

    • Western blot - Anti-KMT5A / SETD8 / Pr-SET7 antibody [EPR24381-24] - BSA and Azide free (AB300518)
      Western blot - Anti-KMT5A / SETD8 / Pr-SET7 antibody [EPR24381-24] - BSA and Azide free (AB300518)
      All lanes : Anti-KMT5A / SETD8 / Pr-SET7 antibody [EPR24381-24] (ab300517) at 1/1000 dilution

      Lane 1 : Wild-type HEK-293 (human embryonic kidney epithelial cell) whole cell lysate
      Lane 2 : KMT5A knockout HEK-293 whole cell lysate

      Lysates/proteins at 20 µg per lane.

      Secondary
      All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/20000 dilution

      Observed band size: 43 kDa why is the actual band size different from the predicted?


      Exposure time: 147 seconds


      This data was developed using ab300517, the same antibody clone in a different buffer formulation. 

      Blocking and diluting buffer and concentration: 5% NFDM/TBST

      Performed under reducing conditions.

      In Western blot, abab300517 was shown to bind specifically to N-lysine methyltransferase KMT5A. A band was observed at 43 kDa in wild-type HEK293 cell lysates whereas no signal observed at this size in N-lysine methyltransferase KMT5A knockout cell line ab261858 (knockout cell lysate ab261667).

      Exposure time: 147 seconds

    • Western blot - Anti-KMT5A / SETD8 / Pr-SET7 antibody [EPR24381-24] - BSA and Azide free (AB300518)
      Western blot - Anti-KMT5A / SETD8 / Pr-SET7 antibody [EPR24381-24] - BSA and Azide free (AB300518)
      All lanes : Anti-KMT5A / SETD8 / Pr-SET7 antibody [EPR24381-24] (ab300517) at 1/1000 dilution

      Lane 1 : HCT116 (human colorectal carcinoma epithelial cell), whole cell lysate
      Lane 2 : SH-SY5Y (human neuroblastoma epithelial cell), whole cell lysate
      Lane 3 : U-2 OS (human bone osteosarcoma epithelial cell), whole cell lysate

      Lysates/proteins at 10 µg per lane.

      Secondary
      All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/20000 dilution

      Observed band size: 43 kDa why is the actual band size different from the predicted?



      This data was developed using ab300517, the same antibody clone in a different buffer formulation. 

      Blocking and diluting buffer and concentration: 5% NFDM/TBST

      Lanes 1/2 were developed using a high sensitivity ECL substrate.

      Lysates were freshly made and used immediately to minimize protein degradation.

      Exposure time: Lanes 1-2: 70 seconds Lane 3: 48 seconds

    • Immunoprecipitation - Anti-KMT5A / SETD8 / Pr-SET7 antibody [EPR24381-24] - BSA and Azide free (AB300518)
      Immunoprecipitation - Anti-KMT5A / SETD8 / Pr-SET7 antibody [EPR24381-24] - BSA and Azide free (AB300518)

      This data was developed using ab300517, the same antibody clone in a different buffer formulation.

      KMT5A / SETD8 / Pr-SET7 was immunoprecipitated from 0.35 mg U-2 OS (human bone osteosarcoma epithelial cell) whole cell lysate 10 ug with ab300517 at 1/30 dilution (2ug in 0.35mg lysates). Western blot was performed on the immunoprecipitate using ab300517 at 1/1000 dilution. VeriBlot for IP secondary antibody (HRP)(ab131366) was used at 1/1000 dilution.

      Lane 1: U-2 OS (human bone osteosarcoma epithelial cell) whole cell lysate 10 ug

      Lane 2: ab300517 IP in U-2 OS (human bone osteosarcoma epithelial cell) whole cell lysate

      Lane 3: Rabbit monoclonal IgG (ab172730) instead of ab300517 in U-2 OS whole cell lysate

      Blocking and dilution buffer and concentration: 5% NFDM/TBST.

      Exposure time: 3 minutes

    • Flow Cytometry (Intracellular) - Anti-KMT5A / SETD8 / Pr-SET7 antibody [EPR24381-24] - BSA and Azide free (AB300518)
      Flow Cytometry (Intracellular) - Anti-KMT5A / SETD8 / Pr-SET7 antibody [EPR24381-24] - BSA and Azide free (AB300518)

      This data was developed using ab300517, the same antibody clone in a different buffer formulation.

      Intracellular flow cytometric analysis of 4% paraformaldehyde fixed 90% methanol permeabilized U-2 OS (human bone osteosarcoma epithelial cell) cells labelling KMT5A / SETD8 / Pr-SET7 with ab300517 at 1/50 dilution (1ug) (Red) compared with a Rabbit monoclonal IgG (ab172730) (Black) isotype control and an unlabeled control (cells without incubation with primary antibody and secondary antibody) (Blue). A Goat Anti-Rabbit IgG (Alexa Fluor® 488, ab150081) at 1/2000 dilution was used as the secondary antibody.

    • Anti-KMT5A / SETD8 / Pr-SET7 antibody [EPR24381-24] - BSA and Azide free (AB300518)
      Anti-KMT5A / SETD8 / Pr-SET7 antibody [EPR24381-24] - BSA and Azide free (AB300518)

    プロトコール

    To our knowledge, customised protocols are not required for this product. Please try the standard protocols listed below and let us know how you get on.

    Click here to view the general protocols

    参考文献 (0)

    ab300518 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。

    ab300518 は論文での使用が確認できていません。

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