Product nameAnti-Tet2 antibody
See all Tet2 primary antibodies
DescriptionRabbit polyclonal to Tet2
Tested applicationsSuitable for: IHC-P, IP, ICC/IF, WBmore details
Species reactivityReacts with: Mouse, Human, Common marmoset
Predicted to work with: Dog
Synthetic peptide conjugated to KLH derived from within residues 1 - 100 of Human Tet2.
- This antibody gave a positive signal in MCF7 whole cell lysate. ICC/IF: MCF7 cells.
Storage instructionsShipped 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 bufferPreservative: 0.02% Sodium Azide
Constituents: 1% BSA, PBS, pH 7.4
Concentration information loading...
PurityImmunogen affinity purified
Our Abpromise guarantee covers the use of ab94580 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
|IHC-P||1/100. Perform heat mediated antigen retrieval before commencing with IHC staining protocol.|
|IP||Use at an assay dependent concentration. PubMed: 24077167|
|ICC/IF||Use a concentration of 10 µg/ml.|
|WB||Use a concentration of 1 µg/ml. Detects a band of approximately 224 kDa (predicted molecular weight: 224 kDa). Abcam recommends using milk (2-5%) as the blocking agent.|
FunctionCatalyzes the conversion of methylcytosine (5mC) to 5-hydroxymethylcytosine (hmC). Plays an important role in myelopoiesis. The clear function of 5-hydroxymethylcytosine (hmC) is still unclear but it may influence chromatin structure and recruit specific factors or may constitute an intermediate component in cytosine demethylation.
Tissue specificityBroadly expressed. Highly expressed in hematopoietic cells; highest expression observed in granulocytes. Expression is reduced in granulocytes from peripheral blood of patients affected by myelodysplastic syndromes.
Involvement in diseaseNote=TET2 is frequently mutated in myeloproliferative disorders (MPD). These constitute a heterogeneous group of disorders, also known as myeloproliferative diseases or myeloproliferative neoplasms (MPN), characterized by cellular proliferation of one or more hematologic cell lines in the peripheral blood, distinct from acute leukemia. Included diseases are: essential thrombocythemia, polycythemia vera, primary myelofibrosis (chronic idiopathic myelofibrosis). Bone marrow samples from patients display uniformly low levels of hmC in genomic DNA compared to bone marrow samples from healthy controls as well as hypomethylation relative to controls at the majority of differentially methylated CpG sites.
Defects in TET2 are a cause of polycythemia vera (PV) [MIM:263300]. A myeloproliferative disorder characterized by abnormal proliferation of all hematopoietic bone marrow elements, erythroid hyperplasia, an absolute increase in total blood volume, but also by myeloid leukocytosis, thrombocytosis and splenomegaly.
Note=TET2 is frequently mutated in systemic mastocytosis; also known as systemic mast cell disease. A condition with features in common with myeloproliferative diseases. It is a clonal disorder of the mast cell and its precursor cells. The clinical symptoms and signs of systemic mastocytosis are due to accumulation of clonally derived mast cells in different tissues, including bone marrow, skin, the gastrointestinal tract, the liver, and the spleen.
Note=TET2 is frequently mutated in myelodysplastic syndromes, a heterogeneous group of closely related clonal hematopoietic disorders. All are characterized by a hypercellular or hypocellular bone marrow with impaired morphology and maturation, dysplasia of the myeloid, megakaryocytic and/or erythroid lineages, and peripheral blood cytopenias resulting from ineffective blood cell production. Included diseases are: refractory anemia (RA), refractory anemia with ringed sideroblasts (RARS), refractory anemia with excess blasts (RAEB), refractory cytopenia with multilineage dysplasia and ringed sideroblasts (RCMD-RS). Chronic myelomonocytic leukemia (CMML) is a myelodysplastic/myeloproliferative disease. Myelodysplastic syndromes are considered a premalignant condition in a subgroup of patients that often progresses to acute myeloid leukemia (AML). Bone marrow samples from patients display uniformly low levels of hmC in genomic DNA compared to bone marrow samples from healthy controls as well as hypomethylation relative to controls at the majority of differentially methylated CpG sites.
Sequence similaritiesBelongs to the TET family.
- Information by UniProt
- FLJ20032 antibody
- KIAA1546 antibody
- MDS antibody
Lane 1: Wild-type HAP1 whole cell lysate (20 µg)
Lane 2: Tet2 knockout HAP1 whole cell lysate (20 µg)
Lane 3: HeLa whole cell lysate (20 µg)
Lane 4: F9 whole cell lysate (20 µg)
Lanes 1 - 4: Merged signal (red and green). Green - ab94580 observed at 250 kDa. Red - loading control, ab18058, observed at 130 kDa.
ab94580 was shown to specifically recognize Tet2 in wild-type HAP1 cells along with additional cross-reactive bands. No band was observed when Tet2 knockout samples were examined. Wild-type and Tet2 knockout samples were subjected to SDS-PAGE. Ab94580 and ab18058 (Mouse anti-Vinculin loading control) were incubated overnight at 4°C at 1 µg/ml and 1/10,000 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 1/10,000 dilution for 1 hour at room temperature before imaging.
Lane 1 : Anti-Tet2 antibody (ab94580) at 1 µg/ml (blocked with 5% BSA)
Lane 2 : Anti-Tet2 antibody (ab94580) at 1 µg/ml (blocked with 5% milk)
Lane 3 : Anti-Tet2 antibody (ab94580) at 1 µg/ml (blocked with 2% milk)
All lanes : MCF7 (Human breast adenocarcinoma cell line) Whole Cell Lysate
Lysates/proteins at 20 µg per lane.
All lanes : Goat Anti-Rabbit IgG H&L (HRP) preadsorbed (ab97080) at 1/5000 dilution
Developed using the ECL technique.
Performed under reducing conditions.
Predicted band size: 224 kDa
Observed band size: 224 kDa
Additional bands at: 300 kDa, 55 kDa, 70 kDa. We are unsure as to the identity of these extra bands.
Exposure time: 20 minutes
Based on this data we recommend using milk as the blocking agent. We welcome customer feedback and would appreciate any comments regarding this product and the data presented above.
ICC/IF image of ab94580 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 ab94580 at 10µg/ml overnight at +4°C. The secondary antibody (green) was DyLight® 488 goat anti- rabbit (ab96899) 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 HeLa pfa fixed (4%, 10 minutes) cell types at 10ug/ml.
ab94580 staining Tet2 in Marmoset adult testis tissue sections by Immunohistochemistry (IHC-P - paraformaldehyde-fixed, paraffin-embedded sections). Tissue was fixed with formaldehyde and blocked with 5% milk for 30 minutes at 25°C; antigen retrieval was by heat mediation in Dako antigen retrieval solution. Samples were incubated with primary antibody (1/100 in 5% milk) for 18 hours at 4°C. An Alexa Fluor® 555-conjugated Goat anti-rabbit polyclonal (1/500) was used as the secondary antibody.
This product has been referenced in:
- Ma M et al. Preeclampsia is associated with hypermethylation of IGF-1 promoter mediated by DNMT1. Am J Transl Res 10:16-39 (2018). IHC-P ; Human . Read more (PubMed: 29422991) »
- Yang R et al. Tet1 and Tet2 maintain mesenchymal stem cell homeostasis via demethylation of the P2rX7 promoter. Nat Commun 9:2143 (2018). Read more (PubMed: 29858571) »