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使用敲除细胞株进行验证

Anti-KDM5A / Jarid1A / RBBP2抗体[18E8] (ab78322)

Submit a review Submit a question References (11)
Western blot - Anti-KDM5A / Jarid1A / RBBP2 antibody [18E8] (ab78322)
  • Immunocytochemistry/ Immunofluorescence - Anti-KDM5A / Jarid1A / RBBP2 antibody [18E8] (ab78322)
  • Western blot - Anti-KDM5A / Jarid1A / RBBP2 antibody [18E8] (ab78322)
  • Flow Cytometry (Intracellular) - Anti-KDM5A / Jarid1A / RBBP2 antibody [18E8] (ab78322)

Key features and details

  • Mouse monoclonal [18E8] to KDM5A / Jarid1A / RBBP2
  • Suitable for: Flow Cyt (Intra), ICC/IF, WB
  • Knockout validated
  • Reacts with: Mouse, Human
  • Isotype: IgG2a

概述

  • 产品名称

    Anti-KDM5A / Jarid1A / RBBP2抗体[18E8]
    参阅全部 KDM5A / Jarid1A / RBBP2 一抗

  • 描述

    小鼠单克隆抗体[18E8] to KDM5A / Jarid1A / RBBP2

  • 宿主

    Mouse

  • 经测试应用

    适用于: Flow Cyt (Intra), ICC/IF, WBmore details

  • 种属反应性

    与反应: Mouse, Human

  • 免疫原

    Synthetic peptide corresponding to Human KDM5A/ Jarid1A/ RBBP2 aa 1416-1434.
    Sequence:

    PRKQPRKSPLVPRSLEPPV


    Database link: P29375
    Run BLAST with BLAST the sequence with ExPASy Run BLAST with BLAST the sequence with NCBI

  • 阳性对照

    • HeLa siRNA. HeLa RBP2 siRNA. Extracts from MCF7 cells, U2OS cells, NIH3T3 cells and J1 (mouse ES) cells. HeLa cells : Flow Cyt (Intra).

  • 常规说明

    Histone demethylase that specifically demethylates 'Lys-4' of histone H3, thereby playing a central role in histone code.

    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

应用

The Abpromise guarantee

Abpromise™承诺保证使用ab78322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
Flow Cyt (Intra)
Use 1µg for 106 cells.

ab170191 - Mouse monoclonal IgG2a, is suitable for use as an isotype control with this antibody.
ICC/IF
Use a concentration of 1 - 10 µg/ml.
WB
Use a concentration of 1 µg/ml. Predicted molecular weight: 196 kDa.
说明
Flow Cyt (Intra)
Use 1µg for 106 cells.

ab170191 - Mouse monoclonal IgG2a, is suitable for use as an isotype control with this antibody.
ICC/IF
Use a concentration of 1 - 10 µg/ml.
WB
Use a concentration of 1 µg/ml. Predicted molecular weight: 196 kDa.

靶标

  • 功能

    Histone demethylase that specifically demethylates 'Lys-4' of histone H3, thereby playing a central role in histone code. Does not demethylate histone H3 'Lys-9', H3 'Lys-27', H3 'Lys-36', H3 'Lys-79' or H4 'Lys-20'. Demethylates trimethylated and dimethylated but not monomethylated H3 'Lys-4'. May stimulate transcription mediated by nuclear receptors. May be involved in transcriptional regulation of Hox proteins during cell differentiation. May participate in transcriptional repression of cytokines such as CXCL12.

  • 序列相似性

    Belongs to the JARID1 histone demethylase family.
    Contains 1 ARID domain.
    Contains 1 JmjC domain.
    Contains 1 JmjN domain.
    Contains 3 PHD-type zinc fingers.

  • 结构域

    The GSGFP motif is required for the interaction with SUZ12.

  • 细胞定位

    Nucleus > nucleolus. Occupies promoters of genes involved in RNA metabolism and mitochondrial function.
  • Information by UniProt

  • 数据库链接

    • 别名

      • Histone demethylase JARID1A antibody
      • JARID1A antibody
      • Jumonji/ARID domain containing protein 1A antibody
      • Jumonji/ARID domain-containing protein 1A antibody
      • Kdm5a antibody
      • KDM5A_HUMAN antibody
      • Lysine-specific demethylase 5A antibody
      • RBBP-2 antibody
      • RBBP2 antibody
      • RBP2 antibody
      • Retinoblastoma binding protein 2 antibody
      • Retinoblastoma-binding protein 2 antibody
      see all

    图片

    • Western blot - Anti-KDM5A / Jarid1A / RBBP2 antibody [18E8] (ab78322)
      Western blot - Anti-KDM5A / Jarid1A / RBBP2 antibody [18E8] (ab78322)

      Lane 1: Wild-type HAP1 whole cell lysate (20 µg)
      Lane 2: KDM5A knockout HAP1 whole cell lysate (20 µg)
      Lane 3: HeLa whole cell lysate (20 µg)
      Lane 4: HEK293 whole cell lysate (20 µg)

      Lanes 1 - 4: Merged signal (red and green). Green - ab78322 observed at 240 kDa. Red - loading control, ab176560, observed at 50 kDa.

      ab78322 was shown to specifically react with KDM5A in wild type HAP1 cells along with additional cross reactive bands. No bands were observed when KDM5A knockout samples were examined. Ab78322 and ab176560 (Rabbit anti alpha Tubulin loading control) were incubated overnight at 4°C at 1 ug/ml and 1/10000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye® 800CW) preabsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye® 680RD) preabsorbed (ab216776) secondary antibodies at 1/20000 dilution for 1 hour at room temperature before imaging.

       

       

    • Immunocytochemistry/ Immunofluorescence - Anti-KDM5A / Jarid1A / RBBP2 antibody [18E8] (ab78322)
      Immunocytochemistry/ Immunofluorescence - Anti-KDM5A / Jarid1A / RBBP2 antibody [18E8] (ab78322)

      ab78322 staining KDM5A / Jarid1A / RBBP2 [18E8] in HeLa cell line by ICC/IF (Immunocytochemistry/immunofluorescence). Cells were fixed with 4 % paraformaldehyde, permeabilized with Triton X-100 0.25% in PBS and blocked with 1.5% BSA for 30 minutes. Samples were incubated with primary antibody (1/200 in PBS + 1% BSA) for overnight at 4°C. An Alexa Fluor®488-conjugated Goat anti-mouse IgG polyclonal (1/1000 in PBS + 1% BSA) was used as the secondary antibody and incubated for 1 hour at room temperature. DAPI was used to stain the nuclear DNA.

    • Western blot - Anti-KDM5A / Jarid1A / RBBP2 antibody [18E8] (ab78322)
      Western blot - Anti-KDM5A / Jarid1A / RBBP2 antibody [18E8] (ab78322)
      All lanes : Anti-KDM5A / Jarid1A / RBBP2 antibody [18E8] (ab78322) at 1 µg/ml

      Lane 1 : HeLa control siRNA
      Lane 2 : HeLa RBBP2 siRNA
      Lane 3 : Extracts from MCF7 cells
      Lane 4 : Extracts from U2OS cells
      Lane 5 : Extracts from NIH3T3 cells
      Lane 6 : Extracts from J1 (mouse ES) cells

      Developed using the ECL technique.

      Predicted band size: 196 kDa
      Observed band size: 196 kDa

    • Flow Cytometry (Intracellular) - Anti-KDM5A / Jarid1A / RBBP2 antibody [18E8] (ab78322)
      Flow Cytometry (Intracellular) - Anti-KDM5A / Jarid1A / RBBP2 antibody [18E8] (ab78322)
      Overlay histogram showing HeLa cells stained with ab78322 (red line). The cells were fixed with 80% methanol (5 min) and then permeabilized with 0.1% PBS-Tween for 20 min. The cells were then incubated in 1x PBS / 10% normal goat serum / 0.3M glycine to block non-specific protein-protein interactions followed by the antibody (ab78322, 1µg/1x106 cells) for 30 min at 22ºC. The secondary antibody used was DyLight® 488 goat anti-mouse IgG (H+L) (ab96879) at 1/500 dilution for 30 min at 22ºC. Isotype control antibody (black line) was mouse IgG2a [ICIGG2A] (ab91361, 1µg/1x106 cells) used under the same conditions. Acquisition of >5,000 events was performed.

    文献 (11)

    发表研究结果有使用 ab78322？请让我们知道，以便我们可以引用本数据表中的参考文章。

    ab78322 被引用在 11 文献中.

    • Jo JH  et al. RepID represses megakaryocytic differentiation by recruiting CRL4A-JARID1A at DAB2 promoter. Cell Commun Signal 21:219 (2023). PubMed: 37612584
    • Li S  et al. Fbxo22 inhibits metastasis in triple-negative breast cancer through ubiquitin modification of KDM5A and regulation of H3K4me3 demethylation. Cell Biol Toxicol N/A:N/A (2022). PubMed: 36112263
    • Zhou M  et al. miR-181d/RBP2/NF-?B p65 Feedback Regulation Promotes Chronic Myeloid Leukemia Blast Crisis. Front Oncol 11:654411 (2021). PubMed: 33842368
    • Kumbhar R  et al. Poly(ADP-ribose) binding and macroH2A mediate recruitment and functions of KDM5A at DNA lesions. J Cell Biol 220:N/A (2021). PubMed: 34003252
    • Gong F  et al. Histone demethylase KDM5A regulates the ZMYND8-NuRD chromatin remodeler to promote DNA repair. J Cell Biol 216:1959-1974 (2017). PubMed: 28572115
    View all Publications for this product

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