Anti-KMT4 / Dot1L抗体(ab64077)

概述

  • 产品名称Anti-KMT4 / Dot1L抗体
    参阅全部 KMT4 / Dot1L 一抗
  • 描述
    兔多克隆抗体to KMT4 / Dot1L
  • 经测试应用适用于: WB, ICC/IF, IHC-Pmore details
  • 种属反应性
    与反应: Mouse, Rat, Human
    预测可用于: Rhesus monkey
  • 免疫原

    Synthetic peptide conjugated to KLH derived from within residues 1700 to the C-terminus of Human KMT4/ Dot1L.

    (Peptide available as ab71337.)

  • 阳性对照
    • This antibody gave a positive signal in Mouse Liver Tissue Lysate, as well as Human and Rat Liver Tissue Lysates (data not shown).

性能

应用

Our Abpromise guarantee covers the use of ab64077 in the following tested applications.

The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.

应用 Ab评论 说明
WB Use a concentration of 2 - 5 µg/ml. Detects a band of approximately 171 kDa (predicted molecular weight: 185 kDa).

Abcam recommends using 1% Milk as the blocking agent.

ICC/IF Use a concentration of 5 µg/ml.
IHC-P Use at an assay dependent concentration. PubMed: 22566624

靶标

  • 功能Histone methyltransferase. Methylates 'Lys-79' of histone H3. Nucleosomes are preferred as substrate compared to free histones. Binds to DNA.
  • 序列相似性Belongs to the class I-like SAM-binding methyltransferase superfamily. DOT1 family.
    Contains 1 DOT1 domain.
  • 细胞定位Nucleus.
  • Information by UniProt
  • 数据库链接
  • 别名
    • Disrupter of telomere silencing protein 1 antibody
    • DOT 1 antibody
    • DOT1 antibody
    • DOT1 like antibody
    • DOT1 like histone H3 methyltransferase (S. cerevisiae) antibody
    • DOT1 like histone H3 methyltransferase antibody
    • DOT1 like histone H3K79 methyltransferase antibody
    • DOT1 like protein antibody
    • DOT1, S. cerevisiae, homolog of antibody
    • DOT1-like protein antibody
    • DOT1L antibody
    • DOT1L_HUMAN antibody
    • EC 2.1.1.43 antibody
    • H3 K79 HMTase antibody
    • H3 lysine-79 specific antibody
    • H3-K79-HMTase antibody
    • Histone H3 K79 methyltransferase antibody
    • Histone H3 methyltransferase antibody
    • Histone H3 methyltransferase DOT1-like antibody
    • Histone H3-K79 methyltransferase antibody
    • Histone lysine N methyltransferase H3 lysine 79 specific antibody
    • Histone methyltransferase DOT1L antibody
    • Histone-lysine N-methyltransferase antibody
    • KIAA1814 antibody
    • KMT4 antibody
    • Lysine N methyltransferase 4 antibody
    • Lysine N-methyltransferase 4 antibody
    see all

Anti-KMT4 / Dot1L antibody 图像

  • Lanes 1 - 2 : Anti-KMT4 / Dot1L antibody (ab64077) at 5 µg/ml (1% Milk Block)
    Lanes 3 - 4 : Anti-KMT4 / Dot1L antibody (ab64077) at 2 µg/ml (1% Milk Block)

    Lane 1 : Human liver tissue lysate - total protein (ab29889)
    Lane 2 : Liver (Mouse) Tissue Lysate
    Lane 3 : Human liver tissue lysate - total protein (ab29889)
    Lane 4 : Liver (Mouse) Tissue Lysate

    Lysates/proteins at 20 µg per lane.

    Secondary
    Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/10000 dilution
    Developed using the ECL technique

    Performed under reducing conditions.

    Predicted band size : 185 kDa
    Observed band size : 170 kDa (why is the actual band size different from the predicted?)
    Additional bands at : 30 kDa,41 kDa,50 kDa,70 kDa. We are unsure as to the identity of these extra bands.

    Exposure time : 20 minutes
  • ICC/IF image of ab64077 stained MCF-7 cells. The cells were 100% Methanol fixed (5 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 (ab64077, 5µg/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. This antibody also gave a positive result in 100% Methanol fixed (5 min) HeLa, Hek293. HepG2 cells at 5µg/ml.

Anti-KMT4 / Dot1L antibody (ab64077)参考文献

This product has been referenced in:
  • Dottermusch-Heidel C  et al. H3K79 methylation directly precedes the histone-to-protamine transition in mammalian spermatids and is sensitive to bacterial infections. Andrology 2:655-65 (2014). IHC-P ; Human . Read more (PubMed: 25079683) »
  • Ontoso D  et al. Dynamics of DOT1L localization and H3K79 methylation during meiotic prophase I in mouse spermatocytes. Chromosoma N/A:N/A (2013). ICC/IF . Read more (PubMed: 24105599) »

See all 4 Publications for this product

Product Wall

For a positive control I was suggesting lysates of testis because that is where the highest expression is. However, if you are confident the protein exists in your kidney samples, then you should try blocking with 1% BSA instead of milk, and diluting t...

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Thank you for the protocol information. I think the two things you should check are a positive control of testes, if available, and the efficacy of the transfer with a Ponceau Red stain of the membrane before incubation with the antibody.

Ex...

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Thank you for contacting us. This is a fairly large protein so good transfer may be the issue, or the membrane blocking conditions or the amount of sample loaded into the gel. I am assuming the problem is that you get no bands. If not, please describe ...

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Thank you for your email. Below please find your discount code for trying the antibody in mouse plusinstructions. After this section is the information about the free of charge replacement.

DISCOUNT CODE: xxx
Expiration date: xxx
<...

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Thank you for your patience.

I have heard back from the lab and for ab72454, the homology with mouse is, unfortunately,not greater than 50%. Thus, the chance of crossreactivity is quite low and I would not recommend trying this antibody in m...

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Thank you for contacting us. I have asked the laboratory that produces ab72454 to align their immunogen sequence with mouse KTM4. I want to be sure that there is high homology before I recommend it as a replacement. If it appears to be sufficient (85% ...

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Thank you for your reply.

As requested I am sending you a vial of ab57827 to try out. You should receive the antibody by 10.30am tomorrow morning. Your new order number is *****.

If there is anything else I can help you with, or if ...

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Thank you for contacting Abcam.

Since ab64077 is covered under our Abpromise to work in western blot on human samples and your data is showing that this current vial is not, I would be happy to send you a replacement antibody, I think ab57827...

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Abcam guarantees this product to work in the species/application used in this Abreview.
Application Immunocytochemistry/ Immunofluorescence
Sample Human Cell (HeLa)
Specification HeLa
Fixative Paraformaldehyde
Permeabilization Yes - 0.5% Triton X100
Username

Dr. Kirk McManus

Verified customer

提交于 Aug 03 2011

Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"