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The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
Use at an assay dependent concentration.
1/250. Detects a band of approximately 223 kDa (predicted molecular weight: 223 kDa).
Use at an assay dependent concentration.
Catalyzes 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.
Broadly 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 disease
Note=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.
All lanes : Anti-Tet2 antibody (ab124297) at 1/250 dilution (Milk blocking - 1%)
Lane 1 : E14Tg2a (Mouse embryonic stem cell line) Whole Cell Lysate Lane 2 : WT Mouse ES Cell Lysate (Positive Control for Tet2) Lane 3 : Tet2 Knockout Mouse ES Cell Lysate (Negative Control)
Lysates/proteins at 25 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 : 223 kDa Observed band size : 223 kDa Additional bands at : 48 kDa (possible non-specific binding),65 kDa (possible non-specific binding),70 kDa (possible non-specific binding).
Exposure time : 4 minutes
This blot was produced using a 3-8% Tris Acetate gel under the TA buffer system. The gel was run at 150V for 60 minutes before being transferred onto a Nitrocellulose membrane at 30V for 70 minutes. The membrane was then blocked for an hour using 1% Milk before being incubated with ab124297 overnight at 4°C. Antibody binding was detected using an anti-rabbit antibody conjugated to HRP, and visualised using ECL development solution.
Western blot - Anti-Tet2 antibody (ab124297)This image is courtesy of an anonymous abreview
All lanes : Anti-Tet2 antibody (ab124297) at 1/1000 dilution (Incubated for 12 hours at 4°C)
Lane 1 : Wild type mouse embryonic stem cells with Milk, 1 hour at 21°C Lane 2 : Tet2 KO clone#1 mouse embryonic stem cells with Milk, 1 hour at 21°C Lane 3 : Tet2 KO clone#2 mouse embryonic stem cells with Milk, 1 hour at 21°C
Lysates/proteins at 20 µg per lane. Blocking peptides at 5 % per lane.
Secondary HRP conjugated Goat anti-rabbit IgG at 1/10000 dilution Developed using the ECL technique
Immunocytochemistry/ Immunofluorescence - Anti-Tet2 antibody (ab124297)This image is courtesy of an anonymous abreview.
ICC/IF image of Anti-Tet2 antibody (ab124297) stained WT and Tet2-/- mouse ES cells. The cells were fixed in PFA, permeabilized using 0.25% Triton X-100, and blocked with 10% BSA for 30 minutes. The cells were then incubated with ab124297 at a 1/400 dilution for 13 hours and 20 minutes at 4oC. The secondary antibody was a Rhodamine Red-X AffiniPure Donkey anti-Rabbit used at a 1/500 dilution.
Immunocytochemistry/ Immunofluorescence - Anti-Tet2 antibody (ab124297)This image is courtesy of an abreview from Joe Segal.
ICC/IF image of Anti-Tet2 antibody (ab124297) stained D3 mouse ES cells. The cells were fixed in 4% PFA, permeabilized using 0.1% Triton X-100, and blocked with 1% Goat serum, 0.1% BSA in PBS for 30 minutes. The cells were then incubated with ab124297 at a 1/100 dilution for 2 hours at RT. The secondary antibody was a Goat polyclonal Secondary Antibody to Rabbit IgG – H&L Alexa Fluor 488 (ab150077) used at a 1/200 dilution.
Immunoprecipitation - Anti-Tet2 antibody (ab124297)This image is courtesy of an anonymous abreview.
Immunoprecipitation of Tet2, using WT and Tet2 KO mouse ES cells. The protein was immunoprecipitated using 300µg whole cell extract, and 3µg of Rabbit polyclonal to Tet2 (ab124297). The same Tet2 antibody was used for the western blot. Tet2 KO mouse ES cells were used as a negative control.
Mishra M & Kowluru RA The Role of DNA Methylation in the Metabolic Memory Phenomenon Associated With the Continued Progression of Diabetic Retinopathy. Invest Ophthalmol Vis Sci57:5748-5757 (2016).
Read more (PubMed: 27787562) »
Lio CW et al. Tet2 and Tet3 cooperate with B-lineage transcription factors to regulate DNA modification and chromatin accessibility. Elife5:N/A (2016).
Read more (PubMed: 27869616) »