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使用敲除细胞株进行验证重组RabMAb

重组Anti-HDAC8抗体[EPR23837-45] - BSA and Azide free (ab274378)

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Western blot - Anti-HDAC8 antibody [EPR23837-45] - BSA and Azide free (ab274378)
  • Immunoprecipitation - Anti-HDAC8 antibody [EPR23837-45] - BSA and Azide free (ab274378)
  • Western blot - Anti-HDAC8 antibody [EPR23837-45] - BSA and Azide free (ab274378)
  • Western blot - Anti-HDAC8 antibody [EPR23837-45] - BSA and Azide free (ab274378)
  • Immunoprecipitation - Anti-HDAC8 antibody [EPR23837-45] - BSA and Azide free (ab274378)
  • Anti-HDAC8 antibody [EPR23837-45] - BSA and Azide free (ab274378)

Key features and details

  • Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
  • Rabbit monoclonal [EPR23837-45] to HDAC8 - BSA and Azide free
  • Suitable for: WB, IP
  • Knockout validated
  • Reacts with: Mouse, Rat, Human

Conjugates logo Related conjugates and formulations

Unconjugated

概述

  • 产品名称

    Anti-HDAC8抗体[EPR23837-45] - BSA and Azide free
    参阅全部 HDAC8 一抗

  • 描述

    兔单克隆抗体[EPR23837-45] to HDAC8 - BSA and Azide free

  • 宿主

    Rabbit

  • 经测试应用

    适用于: WB, IPmore details
    不适用于: Flow Cyt,ICC/IF or IHC-P

  • 种属反应性

    与反应: Mouse, Rat, Human

  • 免疫原

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

  • 阳性对照

    • WB: HAP1, K562, HeLa,LADMAC and C6 whole cell lysates; Human brain tissue lysate; Mouse brain tissue lysate; rat brain tissue lysate. IP: K562 and NIH/3T3 whole cell lysates.

  • 常规说明

    ab274378 is the carrier-free version of ab274372.

    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.

性能

应用

The Abpromise guarantee

Abpromise™承诺保证使用ab274378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
WB
Use at an assay dependent concentration. Detects a band of approximately 42 kDa (predicted molecular weight: 42 kDa).
IP
Use at an assay dependent concentration.
说明
WB
Use at an assay dependent concentration. Detects a band of approximately 42 kDa (predicted molecular weight: 42 kDa).
IP
Use at an assay dependent concentration.
应用说明
Is unsuitable for Flow Cyt,ICC/IF or IHC-P.

靶标

  • 功能

    Responsible for the deacetylation of lysine residues on the N-terminal part of the core histones (H2A, H2B, H3 and H4). Histone deacetylation gives a tag for epigenetic repression and plays an important role in transcriptional regulation, cell cycle progression and developmental events. Histone deacetylases act via the formation of large multiprotein complexes. May play a role in smooth muscle cell contractility.

  • 组织特异性

    Weakly expressed in most tissues. Expressed at higher level in heart, brain, kidney and pancreas and also in liver, lung, placenta, prostate and kidney.

  • 序列相似性

    Belongs to the histone deacetylase family. HD type 1 subfamily.

  • 翻译后修饰

    Phosphorylated by PKA on serine 39. Phosphorylation reduces deacetylase activity observed preferentially on histones H3 and H4.

  • 细胞定位

    Nucleus. Cytoplasm. Excluded from the nucleoli. Found in the cytoplasm of cells showing smooth muscle differentiation.
  • Information by UniProt

  • 数据库链接

    see all

  • 别名

    • CDA07 antibody
    • CDLS5 antibody
    • HD 8 antibody
    • HD8 antibody
    • HDAC 8 antibody
    • Hdac8 antibody
    • HDAC8_HUMAN antibody
    • HDACL 1 antibody
    • HDACL1 antibody
    • Histone deacetylase 8 antibody
    • Histone deacetylase like 1 antibody
    • MRXS6 antibody
    • RPD 3 antibody
    • RPD3 antibody
    • WTS antibody
    see all

图片

  • Western blot - Anti-HDAC8 antibody [EPR23837-45] - BSA and Azide free (ab274378)
    Western blot - Anti-HDAC8 antibody [EPR23837-45] - BSA and Azide free (ab274378)
    All lanes : Anti-HDAC8 antibody [EPR23837-45] (ab274372) at 1/1000 dilution

    Lane 1 : Wild-type HAP1 whole cell lysate
    Lane 2 : HDAC8 knockout HAP1 whole cell lysate
    Lane 3 : Mouse brain tissue lysate
    Lane 4 : Rat brain tissue lysate

    Lysates/proteins at 20 µg per lane.

    Secondary
    All lanes : Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated (ab97051) at 1/50000 dilution

    Predicted band size: 42 kDa
    Observed band size: 42 kDa



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

    Ab274372 was shown to specifically react with HDAC8 in wild-type HAP1 cells as signal was lost in HDAC8 knockout cells. Wild-type and HDAC8 knockout samples were subjected to SDS-PAGE. ab274372 and ab129002 (Rabbit anti-Vinculin loading control) were incubated 1 hour at room temperature at 1/1000 dilution and 1/5000 dilution respectively. Blots were developed with Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated (ab97051) secondary antibody at 1/50, 000 dilution for 1 hour at room temperature before imaging. The blot was developed on a BIO-RAD® ChemiDoc™MP instrument using the ECL technique.

    Exposure time: 3 minutes.

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab274372)

  • Immunoprecipitation - Anti-HDAC8 antibody [EPR23837-45] - BSA and Azide free (ab274378)
    Immunoprecipitation - Anti-HDAC8 antibody [EPR23837-45] - BSA and Azide free (ab274378)

    HDAC8 was immunoprecipitated from 0.35 mg NIH/3T3 (mouse embryonic fibroblast) whole cell lysate with ab274372 at 1/30 dilution (2ug in 0.35mg lysates). Western blot was performed on the immunoprecipitate using ab274372 at 1/1000 dilution. VeriBlot for IP Detection Reagent (HRP)(ab131366) was used at 1/5000 dilution.

    Lane 1: NIH/3T3 (mouse embryonic fibroblast) whole cell lysate 10 ug.

    Lane 2: ab274372 IP in NIH/3T3 whole cell lysate.

    Lane 3: Rabbit monoclonal IgG (ab172730) instead of ab274372 in NIH/3T3 whole cell lysate.

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

    Exposure time: 3 minutes.

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab274372).

  • Western blot - Anti-HDAC8 antibody [EPR23837-45] - BSA and Azide free (ab274378)
    Western blot - Anti-HDAC8 antibody [EPR23837-45] - BSA and Azide free (ab274378)
    Anti-HDAC8 antibody [EPR23837-45] (ab274372) at 1/1000 dilution + Human brain tissue lysate at 20 µg

    Secondary
    VeriBlot for IP secondary antibody(HRP)(ab131366) at 1/1000 dilution

    Predicted band size: 42 kDa
    Observed band size: 42 kDa



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

    This blot was developed using a higher sensitivity ECL substrate.

    Exposure time: 3 minutes

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab274372)

  • Western blot - Anti-HDAC8 antibody [EPR23837-45] - BSA and Azide free (ab274378)
    Western blot - Anti-HDAC8 antibody [EPR23837-45] - BSA and Azide free (ab274378)
    All lanes : Anti-HDAC8 antibody [EPR23837-45] (ab274372) at 1/1000 dilution

    Lane 1 : K-562 (human chronic myelogenous leukemia lymphoblast) whole cell lysate
    Lane 2 : HeLa (human cervix adenocarcinoma epithelial cell) whole cell lysate
    Lane 3 : LADMAC (mouse bone marrow monocyte macrophage) whole cell lysate
    Lane 4 : C6 (rat glial tumor glial cell) whole cell lysate

    Lysates/proteins at 20 µg per lane.

    Secondary
    All lanes : Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated (ab97051) at 1/50000 dilution

    Predicted band size: 42 kDa
    Observed band size: 42 kDa



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

    Exposure time: 3 minutes.

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab274372)

  • Immunoprecipitation - Anti-HDAC8 antibody [EPR23837-45] - BSA and Azide free (ab274378)
    Immunoprecipitation - Anti-HDAC8 antibody [EPR23837-45] - BSA and Azide free (ab274378)

    HDAC8 was immunoprecipitated from 0.35 mg K-562 (human chronic myelogenous leukemia lymphoblast) whole cell lysate with ab274372 at 1/30 dilution (2ug in 0.35mg lysates). Western blot was performed on the immunoprecipitate using ab274372 at 1/1000 dilution. VeriBlot for IP Detection Reagent (HRP)(ab131366) was used at 1/5000 dilution.

    Lane 1: K-562 (human chronic myelogenous leukemia lymphoblast) whole cell lysate 10 ug.

    Lane 2: ab274372 IP in K-562 whole cell lysate.

    Lane 3: Rabbit monoclonal IgG (ab172730) instead of ab274372 in K-562 whole cell lysate

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

    Exposure time: 3 minutes.

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab274372).

  • Anti-HDAC8 antibody [EPR23837-45] - BSA and Azide free (ab274378)
    Anti-HDAC8 antibody [EPR23837-45] - BSA and Azide free (ab274378)

实验方案

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

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