电话:
+86 (0512) 65829739
传真:
+86 (010) 6788 5057
电子邮件:
orders@antibodies-online.cn

HDAC1 抗体

This anti-HDAC1 antibody is a 兔 多克隆 antibody detecting HDAC1 in WB, IF, IP, ICC, IHC (p), ChIP, IHC (fro) 和 IHC (wm). Suitable for 人. Independently validated for use in Cleavage Under Targets and Release Using Nuclease. This Primary Antibody has been cited in 7+ publications.
产品编号 ABIN2854776
发货至: 中国

Quick Overview for HDAC1 抗体 (ABIN2854776)

抗原

See all HDAC1 抗体
HDAC1 (Histone Deacetylase 1 (HDAC1))

适用

  • 169
  • 79
  • 64
  • 9
  • 9
  • 7
  • 7
  • 6
  • 6
  • 4
  • 3
  • 2
  • 1
  • 1

宿主

  • 136
  • 39
  • 2

克隆类型

  • 124
  • 53
多克隆

标记

  • 133
  • 8
  • 7
  • 7
  • 3
  • 3
  • 2
  • 2
  • 2
  • 2
  • 2
  • 2
  • 2
  • 2
This HDAC1 antibody is un-conjugated

应用范围

  • 133
  • 78
  • 72
  • 46
  • 37
  • 36
  • 24
  • 24
  • 8
  • 4
  • 3
  • 3
  • 2
  • 2
  • 2
  • 1
  • 1
  • 1
  • 1
  • 1
  • 1
  • 1
  • 1
Western Blotting (WB), Immunofluorescence (IF), Immunoprecipitation (IP), Immunocytochemistry (ICC), Immunohistochemistry (Paraffin-embedded Sections) (IHC (p)), Chromatin Immunoprecipitation (ChIP), Immunohistochemistry (Frozen Sections) (IHC (fro)), Immunohistochemistry (Whole Mount) (IHC (wm))

质量等级

KO Validated
  • 交叉反应

    人, 小鼠, 大鼠, 斑马鱼

    产品特性

    Rabbit Polyclonal antibody to HDAC1 (histone deacetylase 1)
    HDAC1 antibody

    纯化方法

    Purified by antigen-affinity chromatography.

    免疫原

    Recombinant protein encompassing a sequence within the center region of human HDAC1. The exact sequence is proprietary.

    亚型

    IgG
  • 应用备注

    WB: 1:500-1:3000. ICC/IF: 1:100-1:1000. IHC-P: 1:100-1:1000. IP: 1:100-1:500. Optimal dilutions/concentrations should be determined by the researcher. Not tested in other applications.

    说明

    Positive Control: 293T , A431 , HeLa , HepG2 , U87-MG , SK-N-SH , Rat-2 , NIH3T3 , DDDDK-tagged HDAC1-transfected 293T

    Validation: KO/KD, Orthogonal, Overexpression

    限制

    仅限研究用
  • 生效 #104404 (Cleavage Under Targets and Release Using Nuclease)
    '独立验证'标志
    by
    Gianluca Zambanini, Anna Nordin and Claudio Cantù; Cantù Lab, Gene Regulation during Development and Disease, Linköping University
    No.
    #104404
    日期
    2022.02.28
    抗原
    HDAC1
    Lot Number
    Method validated
    Cleavage Under Targets and Release Using Nuclease
    Positive Control

    Polyclonal rabbit anti-H3K4me (antibodies-online, ABIN3023251)

    Negative Control

    Polyclonal guinea pig anti-rabbit IgG (antibodies-online, ABIN101961)

    Notes

    Passed. ABIN2854776 allows for HDAC1 targeted digestion using CUT&RUN in mouse fore limbs (11.5) cells.

    '独立验证'标志
    Validation Images
    Full Methods
    Primary Antibody
    ABIN2854776
    Secondary Antibody
    Full Protocol
    • Cell harvest and nuclear extraction
      • Dissect 3 Fore limbs (11.5 DAC) from mouse strain RjOrl:SWISS for each sample.
      • Dissociate the tissue into single cells in TrypLE for 15 min at 37 °C.
      • Centrifuge cell solution 5 min at 800 x g at RT.
      • Remove the liquid carefully.
      • Gently resuspend cells in 1 mL of Nuclear Extraction Buffer (20 mM HEPES-KOH pH 8.2, 20% Glycerol, 0,05% IGEPAL, 0.5 mM Spermidine, 10 mM KCl, Roche Complete Protease Inhibitor EDTA-free).
      • Move the solution to a 2 mL centrifuge tube.
      • Pellet the nuclei 800 x g for 5 min.
      • Repeat the NE wash twice for a total of three washes.
      • Resuspend the nuclei in 20 µL NE Buffer per sample.
    • Concanavalin A beads preparation
      • Prepare one 2 mL microcentrifuge tube.
      • Gently resuspend the magnetic Concanavalin A Beads (antibodies-online, ABIN6952467).
      • Pipette 20 µL Con A Beads slurry for each sample into the 2 mL microcentrifuge tube.
      • Place the tube on a magnet stand until the fluid is clear. Remove the liquid carefully.
      • Remove the microcentrifuge tube from the magnetic stand.
      • Pipette 1 mL Binding Buffer (20 mM HEPES pH 7.5, 10 mM KCl, 1 mM CaCl2, 1 mM MnCl2) into the tube and resuspend ConA beads by gentle pipetting.
      • Spin down the liquid from the lid with a quick pulse in a table-top centrifuge.
      • Place the tubes on a magnet stand until the fluid is clear. Remove the liquid carefully.
      • Remove the microcentrifuge tube from the magnetic stand.
      • Repeat the wash twice for a total of three washes.
      • Gently resuspend the ConA Beads in a volume of Binding Buffer corresponding to the original volume of bead slurry, i.e. 20 µL per sample.
    • Nuclei immobilization – binding to Concanavalin A beads
      • Carefully vortex the nuclei suspension and add 20 µL of the Con A beads in Binding Buffer to the cell suspension for each sample.
      • Close tube tightly incubates 10 min at 4 °C.
      • Put the 2 mL tube on the magnet stand and when the liquid is clear remove the supernatant.
      • Resuspend the beads in 1 mL of EDTA Wash buffer (20 mM HEPES pH 7.5, 150 mM NaCl, 0.5 mM Spermidine, Roche Complete Protease Inhibitor EDTA-free, 2mM EDTA).
      • Incubate 5 min at RT.
      • Place the tube on the magnet stand and when the liquid is clear remove the supernatant.
      • Resuspend the beads in 200 µl of Wash Buffer (20 mM HEPES pH 7.5, 150 mM NaCl, 0.5 mM Spermidine, Roche Complete Protease Inhibitor EDTA-free) per sample.
    • Primary antibody binding
      • Divide nuclei suspension into separate 200 µL PCR tubes, one for each antibody.
      • Add 2 µL antibody (anti-HDAC1 antibody ABIN2854776, anti-H3K27me3 antibody positive control ABIN6923144, and guinea pig anti-rabbit IgG negative control antibody ABIN101961) to the respective tube, corresponding to a 1:100 dilution.
      • Incubate at 4 °C ON.
      • Place the tubes on a magnet stand until the fluid is clear. Remove the liquid carefully.
      • Remove the microcentrifuge tubes from the magnetic stand.
      • Wash with 200 µL of Wash Buffer using a multichannel pipette to accelerate the process.
      • Repeat the wash five times for a total of six washes.
    • pAG-MNase Binding
      • Prepare a 1.5 mL microcentrifuge tube containing 100 µL of pAG mix per sample (100 µL of wash buffer + 58.5 µg pAG-MNase per sample).
      • Place the PCR tubes with the sample on a magnet stand until the fluid is clear. Remove the liquid carefully.
      • Remove tubes from the magnetic stand.
      • Resuspend the beads in 100 µL of pAG-MNase premix.
      • Incubate 30 min at 4 °C.
      • Place the tubes on a magnet stand until the fluid is clear. Remove the liquid carefully.
      • Remove the microcentrifuge tubes from the magnetic stand.
      • Wash with 200 µL of Wash Buffer using a multichannel pipette to accelerate the process.
      • Repeat the wash five times for a total of six washes.
      • Resuspend in 100 µL of Wash Buffer.
    • MNase digestion and release of pAG-MNase-antibody-chromatin complexes
      • Place PCR tubes on ice and allow to chill.
      • Prepare a 1.5 mL microcentrifuge tube with 102 µl of 2 mM CaCl2 mix per sample (100 µl Wash Buffer + 2 µL 100 mM CaCl2) and let it chill on ice.
      • Always in ice, place the samples on the magnetic rack and when the liquid is clear remove the supernatant.
      • Resuspend the samples in 100 µl of the 2 mM CaCl2 mix and incubate in ice for exactly 30 min.
      • Place the sample on the magnet stand and when the liquid is clear remove the supernatant.
      • Resuspend the sample in 50 µl of 1x Urea STOP Buffer (8.5 M Urea, 100 mM NaCl, 2 mM EGTA, 2 mM EDTA, 0,5% IGEPAL).
      • Incubate the samples 1h at 4°C.
      • Transfer the supernatant containing the pAG-MNase-bound digested chromatin fragments to fresh 200 µl PCR tubes.
    • DNA Clean up
      • Take the Mag-Bind® TotalPure NGS beads (Omega Bio-Tek, M1378-01) from the storage and wait until they are at RT.
      • Add 2x volume of beads to each sample (e.g. 100 µL of beads for 50 µL of sample).
      • Incubate the beads and the sample for 15 min at RT.
      • During incubation prepare fresh EtOH 80%.
      • Place the PCR tubes on a magnet stand and when the liquid is clear remove the supernatant.
      • Add 200 µl of fresh 80% EtOH to the sample without disturbing the beads (Important!!! Do NOT resuspend the beads or remove the tubes from the magnet stand or the sample will be lost).
      • Incubate 30 sec at RT.
      • Remove the EtOH from the sample.
      • Repeat the wash with 80% EtOH.
      • Resuspend the beads in 25 µL of 10 mM Tris.
      • Incubate the sample for 2 min at RT.
      • Repeat the 2x beads clean up as described before (this time with 50 µL of beads for each sample).
      • Resuspend the beads + DNA in 20 µL of 10 mM Tris.
    • Library preparation and sequencing
      • Prepare Libraries using KAPA HyperPrep Kit using KAPA Dual-Indexed adapters according to protocol.
      • Sequence samples on an Illumina NextSeq 500 sequencer, using a NextSeq 500/550 High Output Kit v2.5 (75 Cycles), 36 bp PE.
    • Peak calling
      • Trim reads using using bbTools bbduk (BBMap - Bushnell B. - sourceforge.net/projects/bbmap/) to remove adapters, artifacts and repeat sequences.
      • Map aligned reads to the hg38 human genome using bowtie with options -m 1 -v 0 -I 0 -X 500.
      • Use SAMtools to convert SAM files to BAM files and remove duplicates.
      • Use BEDtools genomecov to produce Bedgraph files.
      • Call peaks using SEACR with a 0.001 threshold and the option norm stringent.
    Experimental Notes

    The protocol is published in Zambanini, G. et al. A New CUT&RUN Low Volume-Urea (LoV-U) protocol uncovers Wnt/β-catenin tissue-specific genomic targets. bioRxiv (2022). https://doi.org/10.1101/2022.07.06.498999

  • 状态

    Liquid

    浓度

    1 mg/mL

    缓冲液

    1XPBS ( pH 7), 20 % Glycerol, 0.01 % Thimerosal

    储存液

    Thimerosal (Merthiolate)

    注意事项

    This product contains Thimerosal (Merthiolate): a POISONOUS AND HAZARDOUS SUBSTANCE which should be handled by trained staff only.

    储存条件

    4 °C,-20 °C

    储存方法

    Store as concentrated solution. Centrifuge briefly prior to opening vial. For short-term storage (1-2 weeks), store at 4°C. For long-term storage, aliquot and store at -20°C or below. Avoid multiple freeze-thaw cycles.
  • Zambanini, Nordin, Jonasson, Pagella, Cantù: "A new cut&run low volume-urea (LoV-U) protocol optimized for transcriptional co-factors uncovers Wnt/b-catenin tissue-specific genomic targets." in: Development (Cambridge, England), (2022) (PubMed).

    Hu, Chung, Ping, Hsu, Tsai, Chen, Cheng: "Differential Expression of Multiple Disease-Related Protein Groups Induced by Valproic Acid in Human SH-SY5Y Neuroblastoma Cells." in: Brain sciences, Vol. 10, Issue 8, (2020) (PubMed).

    Ochiai, Hayashi, Umeda, Yoshimura, Harada, Shimizu, Nakano, Saitoh, Liu, Yamamoto, Okamura, Ohkawa, Kimura, Nikaido: "Genome-wide kinetic properties of transcriptional bursting in mouse embryonic stem cells." in: Science advances, Vol. 6, Issue 25, pp. eaaz6699, (2020) (PubMed).

    Lin, Wang, Wu, Lin, Chen, Chen, Chen, Peng: "Nifedipine Exacerbates Lipogenesis in the Kidney via KIM-1, CD36, and SREBP Upregulation: Implications from an Animal Model for Human Study." in: International journal of molecular sciences, Vol. 21, Issue 12, (2020) (PubMed).

    Deng, Yang, Ji, Lu, Qiu, Sheng, Sun, Kong: "Overexpression of peptidase inhibitor 16 attenuates angiotensin II-induced cardiac fibrosis via regulating HDAC1 of cardiac fibroblasts." in: Journal of cellular and molecular medicine, Vol. 24, Issue 9, pp. 5249-5259, (2020) (PubMed).

    Lee, Lin, Chae, Yoo, Kim, Lee, Johnson, Kim, Cantley, Lee, Yu, Cho: "The chromatin remodeler RSF1 controls centromeric histone modifications to coordinate chromosome segregation." in: Nature communications, Vol. 9, Issue 1, pp. 3848, (2019) (PubMed).

    Lee, Kuo, Tsai, Cheng, Chen, Chan, Lin, Lin, Li, Kanwar, Leung, Cheung, Huang, Wang, Cheung: "Inhibition of HDAC3- and HDAC6-Promoted Survivin Expression Plays an Important Role in SAHA-Induced Autophagy and Viability Reduction in Breast Cancer Cells." in: Frontiers in pharmacology, Vol. 7, pp. 81, (2016) (PubMed).

  • 抗原

    HDAC1 (Histone Deacetylase 1 (HDAC1))

    别名

    histone deacetylase 1

    背景

    Histone acetylation and deacetylation, catalyzed by multisubunit complexes, play a key role in the regulation of eukaryotic gene expression. The protein encoded by this gene belongs to the histone deacetylase/acuc/apha family and is a component of the histone deacetylase complex. It also interacts with retinoblastoma tumor-suppressor protein and this complex is a key element in the control of cell proliferation and differentiation. Together with metastasis-associated protein-2, it deacetylates p53 and modulates its effect on cell growth and apoptosis.

    Cellular Localization: Nucleus

    分子量

    55 kDa

    基因ID

    3065

    UniProt

    Q13547

    途径

    Neurotrophin Signaling Pathway, Intracellular Steroid Hormone Receptor Signaling Pathway, Regulation of Intracellular Steroid Hormone Receptor Signaling, Mitotic G1-G1/S Phases, Regulation of Muscle Cell Differentiation, Skeletal Muscle Fiber Development, Negative Regulation of intrinsic apoptotic Signaling, Embryonic Body Morphogenesis
You are here:
Chat with us!