Anti-KMT4 / Dot1L antibody (ab64077)
Key features and details
- Rabbit polyclonal to KMT4 / Dot1L
- Suitable for: WB, ICC/IF
- Reacts with: Mouse, Human
- Isotype: IgG
Overview
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Product name
Anti-KMT4 / Dot1L antibody
See all KMT4 / Dot1L primary antibodies -
Description
Rabbit polyclonal to KMT4 / Dot1L -
Host species
Rabbit -
Tested applications
Suitable for: WB, ICC/IFmore details -
Species reactivity
Reacts with: Mouse, Human
Predicted to work with: Rat, Rhesus monkey
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Immunogen
Synthetic peptide conjugated to KLH derived from within residues 1700 to the C-terminus of Human KMT4/ Dot1L.
Read Abcam's proprietary immunogen policy (Peptide available as ab71337.) -
Positive control
- WB: Liver (Mouse) Tissue Lysate.
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General notes
Reproducibility is key to advancing scientific discovery and accelerating scientists’ next breakthrough.
Abcam is leading the way with our range of recombinant antibodies, knockout-validated antibodies and knockout cell lines, all of which support improved reproducibility.
We are also planning to innovate the way in which we present recommended applications and species on our product datasheets, so that only applications & species that have been tested in our own labs, our suppliers or by selected trusted collaborators are covered by our Abpromise™ guarantee.
In preparation for this, we have started to update the applications & species that this product is Abpromise guaranteed for.
We are also updating the applications & species that this product has been “predicted to work with,” however this information is not covered by our Abpromise guarantee.
Applications & species from publications and Abreviews that have not been tested in our own labs or in those of our suppliers are not covered by the Abpromise guarantee.
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, as well as customer reviews and Q&As.
Properties
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Form
Liquid -
Storage instructions
Shipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C or -80°C. Avoid freeze / thaw cycle. -
Storage buffer
pH: 7.40
Preservative: 0.02% Sodium azide
Constituent: PBS
Batches of this product that have a concentration
Concentration information loading...Purity
Immunogen affinity purifiedClonality
PolyclonalIsotype
IgGResearch areas
Associated products
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Compatible Secondaries
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Immunizing Peptide (Blocking)
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Isotype control
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Recombinant Protein
Applications
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.
Application Abreviews Notes 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. Target
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Function
Histone methyltransferase. Methylates 'Lys-79' of histone H3. Nucleosomes are preferred as substrate compared to free histones. Binds to DNA. -
Sequence similarities
Belongs to the class I-like SAM-binding methyltransferase superfamily. DOT1 family.
Contains 1 DOT1 domain. -
Cellular localization
Nucleus. - Information by UniProt
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Database links
- Entrez Gene: 84444 Human
- Entrez Gene: 208266 Mouse
- Entrez Gene: 362831 Rat
- Omim: 607375 Human
- SwissProt: Q8TEK3 Human
- Unigene: 713641 Human
- Unigene: 152371 Mouse
- Unigene: 228827 Rat
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Alternative names
- Disrupter of telomere silencing protein 1 antibody
- DOT 1 antibody
- DOT1 antibody
see all
Images
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Western blot - Anti-KMT4 / Dot1L antibody (ab64077)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)
Lanes 1 & 3 : Human liver tissue lysate - total protein (ab29889)
Lanes 2 & 4 : Liver (Mouse) Tissue Lysate
Lysates/proteins at 20 µg per lane.
Secondary
All lanes : 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 -
Immunocytochemistry/ Immunofluorescence - Anti-KMT4 / Dot1L antibody (ab64077)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.
Protocols
Datasheets and documents
References (12)
ab64077 has been referenced in 12 publications.
- Giannios I et al. Protein Methyltransferase Inhibition Decreases Endocrine Specification Through the Upregulation of Aldh1b1 Expression. Stem Cells N/A:N/A (2019). PubMed: 30681750
- Yang L et al. Silencing or inhibition of H3K79 methyltransferase DOT1L induces cell cycle arrest by epigenetically modulating c-Myc expression in colorectal cancer. Clin Epigenetics 11:199 (2019). PubMed: 31888761
- Cornelis FMF et al. Increased susceptibility to develop spontaneous and post-traumatic osteoarthritis in Dot1l-deficient mice. Osteoarthritis Cartilage N/A:N/A (2018). PubMed: 30513362
- Zhang X et al. Prognostic and therapeutic value of disruptor of telomeric silencing-1-like (DOT1L) expression in patients with ovarian cancer. J Hematol Oncol 10:29 (2017). IHC-P ; Human . PubMed: 28114995
- Moretti C et al. SLY regulates genes involved in chromatin remodeling and interacts with TBL1XR1 during sperm differentiation. Cell Death Differ 24:1029-1044 (2017). IF ; Mouse . PubMed: 28475176
Images
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Western blot - Anti-KMT4 / Dot1L antibody (ab64077)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)
Lanes 1 & 3 : Human liver tissue lysate - total protein (ab29889)
Lanes 2 & 4 : Liver (Mouse) Tissue Lysate
Lysates/proteins at 20 µg per lane.
Secondary
All lanes : 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
-
Immunocytochemistry/ Immunofluorescence - Anti-KMT4 / Dot1L antibody (ab64077)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.
