Recombinant Anti-SMYD3 antibody [EPR19311] (ab199361)
![Western blot - Anti-SMYD3 antibody [EPR19311] (ab199361)](https://www.abcam.com/ps/products/199/ab199361/Images/ab199361-369110-anti-smyd3-antibody-epr19311-western-blot-hap1-wt-hap1-smyd3-knockout-hela-mcf7.jpg "Western blot - Anti-SMYD3 antibody [EPR19311] (ab199361)")
Key features and details
- Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
- Rabbit monoclonal [EPR19311] to SMYD3
- Suitable for: WB, IP
- Knockout validated
- Reacts with: Mouse, Rat, Human
Related conjugates and formulations
Overview
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Product name
Anti-SMYD3 antibody [EPR19311]
See all SMYD3 primary antibodies -
Description
Rabbit monoclonal [EPR19311] to SMYD3 -
Host species
Rabbit -
Tested applications
Suitable for: WB, IPmore details -
Species reactivity
Reacts with: Mouse, Rat, Human -
Immunogen
Recombinant fragment. This information is proprietary to Abcam and/or its suppliers.
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Positive control
- WB: Human fetal kidney and fetal brain lysates; wild-type HAP1, HeLa, MCF7, HEK-293, T-47D, HCT 116, LLC, C6, RAW 264.7, PC-12 and NIH/3T3 whole cell lysates; Mouse brain, kidney and spleen lysates; Rat brain and kidney lysates. IP: 293T whole cell lysate.
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General notes
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
Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMAb® patents.
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 long term. Avoid freeze / thaw cycle. -
Storage buffer
pH: 7.2
Preservative: 0.01% Sodium azide
Constituents: 59% PBS, 40% Glycerol (glycerin, glycerine), 0.05% BSA -
Concentration information loading... -
Purity
Protein A purified -
Clonality
Monoclonal -
Clone number
EPR19311 -
Isotype
IgG -
Research areas
Associated products
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Alternative Versions
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Compatible Secondaries
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Isotype control
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Related Products
Applications
The Abpromise guarantee
Our Abpromise guarantee covers the use of ab199361 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 |
1/2000. Detects a band of approximately 45 kDa (predicted molecular weight: 49 kDa).
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| IP |
1/50.
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| Notes |
|---|
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WB
1/2000. Detects a band of approximately 45 kDa (predicted molecular weight: 49 kDa). |
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IP
1/50. |
Target
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Function
Histone methyltransferase. Specifically methylates 'Lys-4' of histone H3, inducing di- and tri-methylation, but not monomethylation. Plays an important role in transcriptional activation as a member of an RNA polymerase complex. Binds DNA containing 5'-CCCTCC-3' or 5'-GAGGGG-3' sequences. -
Tissue specificity
Expressed in skeletal muscles and testis. Overexpressed in a majority of colorectal and hepatocellular carcinomas. -
Sequence similarities
Belongs to the histone-lysine methyltransferase family.
Contains 1 MYND-type zinc finger.
Contains 1 SET domain. -
Cellular localization
Cytoplasm. Nucleus. Mainly cytoplasmic when cells are arrested at G0/G1. Accumulates in the nucleus at S phase and G2/M. - Information by UniProt
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Database links
- Entrez Gene: 64754 Human
- Entrez Gene: 69726 Mouse
- Entrez Gene: 498295 Rat
- Omim: 608783 Human
- SwissProt: Q9H7B4 Human
- SwissProt: Q9CWR2 Mouse
- Unigene: 567571 Human
- Unigene: 222338 Mouse
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Alternative names
- bA74P14.1 (novel protein) antibody
- bA74P14.1 antibody
- FLJ21080 antibody
see all
Images
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Western blot - Anti-SMYD3 antibody [EPR19311] (ab199361)All lanes : Anti-SMYD3 antibody [EPR19311] (ab199361) at 1/1000 dilution
Lane 1 : Wild-type HAP1 whole cell lysate
Lane 2 : SMYD3 knockout HAP1 whole cell lysate
Lane 3 : HeLa whole cell lysate
Lane 4 : MCF7 whole cell lysate
Lysates/proteins at 20 µg per lane.
Performed under reducing conditions.
Predicted band size: 49 kDa
Observed band size: 49 kDaLanes 1 - 4: Merged signal (red and green). Green - ab199361 observed at 49 kDa. Red - loading control, ab8245 (Mouse anti-GAPDH antibody [6C5]) observed at 37kDa.
ab199361 was shown to react with SMYD3 in HAP1 wild-type cells in Western blot. Loss of signal was observed when SMYD3 knockout sample was used. HAP1 wild-type and SMYD3 knockout whole cell lysates were subjected to SDS-PAGE. Membranes were blocked in 3% Milk in TBS-T (0.1% Tween®) before incubation with ab199361 and ab8245 (Mouse anti-GAPDH antibody [6C5]) overnight at 4°C at a 1 in 1000 dilution and a 1 in 20000 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 in 20000 dilution for 1 hour at room temperature before imaging.
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![Western blot - Anti-SMYD3 antibody [EPR19311] (ab199361)](https://www.abcam.com/ps/products/199/ab199361/Images/ab199361-254091-anti-smyd3-antibody-epr19311-western-blot.jpg)
Western blot - Anti-SMYD3 antibody [EPR19311] (ab199361)All lanes : Anti-SMYD3 antibody [EPR19311] (ab199361) at 1/2000 dilution
Lane 1 : Human fetal kidney lysate
Lane 2 : Human fetal brain lysate
Lysates/proteins at 10 µg per lane.
Secondary
All lanes : Goat Anti-Rabbit IgG Peroxidase Conjugate, specific to the non-reduced form of IgG at 1/10000 dilution
Predicted band size: 49 kDa
Observed band size: 45 kDa why is the actual band size different from the predicted?
Exposure time: 3 minutesBlocking/Dilution buffer: 5% NFDM/TBST.
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![Western blot - Anti-SMYD3 antibody [EPR19311] (ab199361)](https://www.abcam.com/ps/products/199/ab199361/Images/ab199361-254090-anti-smyd3-antibody-epr19311-western-blot.jpg)
Western blot - Anti-SMYD3 antibody [EPR19311] (ab199361)All lanes : Anti-SMYD3 antibody [EPR19311] (ab199361) at 1/2000 dilution
Lane 1 : HEK-293 (Human epithelial cell line from embryonic kidney) whole cell lysate
Lane 2 : HeLa (Human epithelial cell line from cervix adenocarcinoma) whole cell lysate
Lane 3 : T-47D (Human ductal breast epithelial tumor cell line) whole cell lysate
Lane 4 : HCT 116 (Human colorectal carcinoma cell line) whole cell lysate
Lane 5 : LLC (Mouse lung carcinoma cell line) whole cell lysate
Lysates/proteins at 20 µg per lane.
Secondary
All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/100000 dilution
Predicted band size: 49 kDa
Observed band size: 45 kDa why is the actual band size different from the predicted?
Exposure time: 3 minutesBlocking/Dilution buffer: 5% NFDM/TBST.
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![Western blot - Anti-SMYD3 antibody [EPR19311] (ab199361)](https://www.abcam.com/ps/products/199/ab199361/Images/ab199361-254088-anti-smyd3-antibody-epr19311-western-blot.jpg)
Western blot - Anti-SMYD3 antibody [EPR19311] (ab199361)All lanes : Anti-SMYD3 antibody [EPR19311] (ab199361) at 1/2000 dilution
Lane 1 : Mouse brain lysate
Lane 2 : Mouse kidney lysate
Lane 3 : Mouse spleen lysate
Lane 4 : Rat brain lysate
Lane 5 : Rat kidney lysate
Lane 6 : C6 (Rat glial tumor cell line) whole cell lysate
Lane 7 : RAW 264.7 (Mouse macrophage cell line transformed with Abelson murine leukemia virus) whole cell lysate
Lane 8 : PC-12 (Rat adrenal gland pheochromocytoma cell line) whole cell lysate
Lane 9 : NIH/3T3 (Mouse embryo fibroblast cell line) whole cell lysate
Lysates/proteins at 10 µg per lane.
Secondary
All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/100000 dilution
Predicted band size: 49 kDa
Observed band size: 45 kDa why is the actual band size different from the predicted?
Exposure time: 3 minutesBlocking/Dilution buffer: 5% NFDM/TBST.
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![Immunoprecipitation - Anti-SMYD3 antibody [EPR19311] (ab199361)](https://www.abcam.com/ps/products/199/ab199361/Images/ab199361-254086-anti-smyd3-antibody-epr19311-immunoprecipitation.jpg)
Immunoprecipitation - Anti-SMYD3 antibody [EPR19311] (ab199361)SMYD3 was immunoprecipitated from 1mg of 293T (Human epithelial cell line from embryonic kidney) whole cell lysate with ab199361 at 1/50 dilution.
Western blot was performed from the immunoprecipitate using ab199361 at 1/1000 dilution.
VeriBlot for IP Detection Reagent (HRP) (ab131366), was used for detection at 1/10000 dilution.
Lane 1: 293T whole cell lysate, 10µg (Input).
Lane 2: ab199361 IP in 293T whole cell lysate.
Lane 3: Rabbit IgG,monoclonal [EPR25A ]-Isotype Control (ab172730) instead of ab199361 in 293T whole cell lysate.
Blocking and dilution buffer and concentration: 5% NFDM/TBST.
Exposure time: 3 minutes.
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![Immunoprecipitation - Anti-SMYD3 antibody [EPR19311] (ab199361)](https://www.abcam.com/ps/products/199/ab199361/Images/ab199361-254084-anti-smyd3-antibody-epr19311-immunoprecipitation.jpg)
Immunoprecipitation - Anti-SMYD3 antibody [EPR19311] (ab199361)SMYD3 was immunoprecipitated from 1mg of 293T (Human epithelial cell line from embryonic kidney) whole cell lysate with ab199361 at 1/50 dilution.
Western blot was performed from the immunoprecipitate using ab199361 at 1/1000 dilution (Panel A) or ab183498 at 1/1000 dilution (Panel B).
VeriBlot for IP Detection Reagent (HRP) (ab131366), was used for detection at 1/10000 dilution.
Lane 1: 293T whole cell lysate, 10µg (Input).
Lane 2: ab199361 IP in 293T whole cell lysate.
Lane 3: Rabbit IgG,monoclonal [EPR25A]-Isotype Control (ab172730) instead of ab199361 in 293T whole cell lysate.
Blocking and dilution buffer and concentration: 5% NFDM/TBST.
Exposure time: 3 minutes.
The product was validated with ab183498, which is specific to SMYD3.
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![Anti-SMYD3 antibody [EPR19311] (ab199361)](https://www.abcam.com/ps/products/199/ab199361/Images/ab199361-8-benefits-of-recombinant-antibodies.png)
Anti-SMYD3 antibody [EPR19311] (ab199361)
Protocols
Datasheets and documents
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Datasheet download
Certificate of Compliance
References (0)
ab199361 has not yet been referenced specifically in any publications.
