Invitrogen

SMYD3 Polyclonal Antibody

Advanced Verification

This Antibody was verified by Knockout to ensure that the antibody binds to the antigen stated.

2 Published Figures

1 Reference

View all (15) SMYD3 antibodies

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Datasheet

Protocols

Questions & Answers

Tech Support

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SMYD3 Antibody in Immunocytochemistry (ICC/IF) — Immunofluorescent analysis of SMYD3 showing staining in the cytoplasm of HCT116 cells. HCT116 cells were fixed in ice-cold MeOH for 5 min and stained using a SMYD3 polyclonal antibody (Product # PA5-31919) diluted at 1:500. Blue: Hoechst 33342 staining.

SMYD3 Antibody in Western Blot (WB) — Western Blot analysis of SMYD3 was performed by separating 30 µg of non-transfected (–) and transfected (+) 293T whole cell extracts by 10% SDS-PAGE. Proteins were transferred to a membrane and probed with a SMYD3 Polyclonal Antibody (Product # PA5-31919) at a dilution of 1:1000. The HRP-conjugated anti-rabbit IgG antibody was used to detect the primary antibody.

SMYD3 Antibody in Immunoprecipitation (IP) — Immunoprecipitation of SMYD3 was performed in 293T whole cell extracts using 5 µg of SMYD3 Polyclonal Antibody (Product # PA5-31919). Samples were transferred to a membrane and probed with SMYD3 Polyclonal Antibody as a primary antibody and an HRP-conjugated anti-Rabbit IgG was used as a secondary antibody.

SMYD3 Antibody in ChIP assay (ChIP) — Cross-linked ChIP analysis of SMYD3 in HepG2 chromatin extract using 5 µg of either normal rabbit IgG or anti-SMYD3 antibody (Product # PA5-31919). The precipitated DNA was detected by PCR with primer set targeting to 5' flanking region of Nkx2.8.

SMYD3 Antibody — Fig 5 Gene ablation techniques show no dependence on SMYD2 or SMYD3 for cancer cell proliferation. Waterfall plot representing LogP RSA scores for sgRNAs targeting A) SMYD2 and B) SMYD3. 313 cell lines were infected with a library of 6500 sgRNAs targeting 600 different genes. LogP RSA scores represent depletion of guides from an infected cell population. Each bar represents a different cell line. Bars are colored by cancer subtype. C) Percent confluency of Hep3B cells infected with CRISPR viruses containing CAS9 and sgRNAs targeting HBE-1, EZH2 (negative controls) or SMYD3. Cell density was evaluated using an Incucyte Zoom. Growth curves were initiated 24 days following virus infection and puromycin selection. Plotted data is the average of three biological replicates. Error bars represent standard deviation (not readily visible on scale). D) SMYD3 western blot of lysates derived from Hep3B cells infected with CAS9 and SMYD3 sgRNA. Parental Hep3Bs and Hep3Bs stably infected with HBE-1, EZH2 (negative controls) or SMYD3 were lysed and probed for SMYD3 levels by western. GAPDH levels were evaluated as a loading control. {KO}

Product Details

PA5-31919

Applications	Tested Dilution	Publications
Western Blot (WB)	1:500-1:3,000	View 1 publication 1 publication
Immunocytochemistry (ICC/IF)	1:100-1:1,000	-
Immunoprecipitation (IP)	1:100-1:500	-
ChIP assay (ChIP)	Assay-dependent	-

Product Specifications
Species Reactivity	Human, Mouse
Published species	Not Applicable
Host/Isotype	Rabbit / IgG
Class	Polyclonal
Type	Antibody
Immunogen	Recombinant protein fragment corresponding to a region within amino acids 22 and 411 of Human SMYD3 View antigen
Conjugate	Unconjugated
Form	Liquid
Concentration	1.23 mg/mL
Purification	Antigen affinity chromatography
Storage buffer	PBS, pH 7, with 20% glycerol
Contains	0.025% ProClin 300
Storage conditions	Store at 4°C short term. For long term storage, store at -20°C, avoiding freeze/thaw cycles.
RRID	AB_2549392

Product Specific Information

Recommended positive controls: DDDDK-tagged SMYD3-transfected 293T, 293T, A431, HeLa, HepG2, A549, SKOV3, HCT116.

Predicted reactivity: Human (99%), Mouse (93%), Rat (94%), Pig (95%), Rhesus Monkey (99%), Bovine (83%).

Store product as a concentrated solution. Centrifuge briefly prior to opening the vial.

Target Information

SMYD3 (histone-lysine N-methyltransferase) methylates Lys-4 of histone H3 and Lys-5 of histone H4. It induces di- and tri-methylation. SMYD3 plays an important role in transcriptional activation as a member of an RNA polymerase complex.

For Research Use Only. Not for use in diagnostic procedures. Not for resale without express authorization.

Product References

Small molecule inhibitors and CRISPR/Cas9 mutagenesis demonstrate that SMYD2 and SMYD3 activity are dispensable for autonomous cancer cell proliferation.

PloS one

Thomenius MJ,Totman J,Harvey D,Mitchell LH,Riera TV,Cosmopoulos K,Grassian AR,Klaus C,Foley M,Admirand EA,Jahic H,Majer C,Wigle T,Jacques SL,Gureasko J,Brach D,Lingaraj T,West K,Smith S,Rioux N,Waters NJ,Tang C,Raimondi A,Munchhof M,Mills JE,Ribich S,Porter Scott M,Kuntz KW,Janzen WP,Moyer M,Smith JJ,Chesworth R,Copeland RA,Boriack-Sjodin PA

Published figure using SMYD3 polyclonal antibody (Product # PA5-31919) in Western Blot

Tue Dec 11 00:00:00 EST 2018

Bioinformatics

Protein Aliases: bA74P14.1 (novel protein); Histone-lysine N-methyltransferase SMYD3; SET and MYND domain-containing protein 3; Zinc finger MYND domain-containing protein 1; zinc finger protein, subfamily 3A (MYND domain containing), 1; zinc finger, MYND domain containing 1

Gene Aliases: 2410008A19Rik; bA74P14.1; KMT3E; SMYD3; ZMYND1; ZNFN3A1

UniProt ID: (Human) Q9H7B4, (Mouse) Q9CWR2

Entrez Gene ID: (Human) 64754, (Mouse) 69726

Molecular Function: histone modifying enzyme