Overview

  • Product name
  • Description
    Rabbit polyclonal to Tet2
  • Host species
    Rabbit
  • Tested applications
    Suitable for: IHC-P, IP, ICC/IF, WBmore details
  • Species reactivity
    Reacts with: Mouse, Human, Common marmoset
    Predicted to work with: Dog
  • Immunogen

    Synthetic peptide conjugated to KLH derived from within residues 1 - 100 of Human Tet2.

    Read Abcam's proprietary immunogen policy

  • Positive control
    • This antibody gave a positive signal in MCF7 whole cell lysate. ICC/IF: MCF7 cells.

Properties

  • 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
    Preservative: 0.02% Sodium Azide
    Constituents: 1% BSA, PBS, pH 7.4
  • Concentration information loading...
  • Purity
    Immunogen affinity purified
  • Clonality
    Polyclonal
  • Isotype
    IgG
  • Research areas

Applications

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.

Application Abreviews Notes
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.

Target

  • Function
    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.
  • Tissue specificity
    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.
  • Sequence similarities
    Belongs to the TET family.
  • Information by UniProt
  • Database links
  • Alternative names
    • FLJ20032 antibody
    • KIAA1546 antibody
    • MDS antibody
    • Methylcytosine dioxygenase TET2 antibody
    • Nbla00191 antibody
    • Probable methylcytosine dioxygenase TET2 antibody
    • Protein Ayu17 449 antibody
    • Tet 2 antibody
    • Tet methylcytosine dioxygenase 2 antibody
    • Tet oncogene 2 antibody
    • Tet oncogene family member 2 antibody
    • TET2 antibody
    • TET2_HUMAN antibody
    see all

Images

  • 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.

    Secondary
    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.

    See Abreview

References

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) »
See all 22 Publications for this product

Customer reviews and Q&As

1-10 of 18 Abreviews or Q&A

Application
Western blot
Sample
Rat Cell lysate - whole cell (HEK and NRK)
Gel Running Conditions
Reduced Denaturing (6)
Loading amount
20 µg
Specification
HEK and NRK
Blocking step
(agent) for 1 hour(s) and 0 minute(s) · Concentration: 5µg/mL · Temperature: RT°C

Abcam user community

Verified customer

Submitted Apr 28 2017

Application
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Blocking step
Milk as blocking agent for 30 minute(s) · Concentration: 5% · Temperature: 25°C
Antigen retrieval step
Heat mediated - Buffer/Enzyme Used: Dako antigen retrieval solution
Sample
Marmoset (common) Tissue sections (Adult testis)
Specification
Adult testis
Permeabilization
No
Fixative
Formaldehyde

Dr. 煜欽 林

Verified customer

Submitted Jan 20 2014

Answer

Thank you for sharing those results. It is disappointing to hear that this new lot has not provided a better result.
I have raised this problem with you lab and quality control team. We are currently trying to ascertain why this change in reactivity has been observed with this antibody with the two most recent lots you have tried. And will be initiating further in house testing to try and understand why this problem may have occurred.
I am sorry for the inconvenience this problem has caused you. I am sorry but I do not have any other antibody I am able to offer which has been tested in ICC as yet, and until I am confident we have resolved the problem with the staining seen with ab94580 I don't want to provide you with another lot of this. You may want to have a look at the ab127416 which has been used in staining of paraffin embedded human bladder but I would not be able to guarantee how it would perform in ICC.
Alternatively, if you would prefer I can provide you with any other antibody from our catalogue as a replacement, or a refund or credit note.
I look forward to receiving your reply.

Read More
Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Immunocytochemistry/ Immunofluorescence
Sample
Mouse Cell (mouse pluripotent cells)
Specification
mouse pluripotent cells
Fixative
Paraformaldehyde
Permeabilization
Yes - 0.25% Triton X-100
Blocking step
BSA as blocking agent for 30 minute(s) · Concentration: 10% · Temperature: 37°C

Abcam user community

Verified customer

Submitted Nov 02 2012

Answer

I am sorry for the delay in getting back to you.
I can now confirm that we do have a different lot of ab94580 in stock which has been tested in both Western blotting and IF. If you would like I can arrange to send this to you. In order to arrange this, could you provide me with the order number on which ab94580 was purchased (the delivery address used)?
Alternatively, if you would prefer, I can arrange for a different antibody altogether to be sent, or arrange for a credit note or refund.
I look forward to receiving your reply.

Read More

Answer

Thank you very much for that reply.
That information has been very helpful in looking into this case further. I am sorry that the particular lot has not been producing the expected results.
I am currently looking into whether we have a different lot number to offer, and how this has been evaluated. I will get back to you as soon as I have confirmation of this.
In order to help understand the difference in reactivity you have observed, would you mind sending a few images? Particularly, the Western blot achieved with each of the lots tried as well as the IF images?
Many thanks for your continued cooperation.

Read More

Answer

Thank you for contacting us and reporting the problems you have been encountering with the Anti-Tet2 antibody (ab94580).
I am sorry to hear you are experiencing difficulties with one of our products. We take product complaints very seriously, and investigate every product that we feel may not be performing correctly. I have checked in our stock and unfortunately we do not currently have any more stock for the lot number GR51556-1 available. However, I would like to investigate this problem a little further to see if we can identify a reason for this difference and it may be that I am able to offer you a different lot altogether.
To this end, I would like to gather a little more information from you in regards to the samples tested, the protocol used and the results observed. I have therefore attached a questionnaire. This should only take 5-10 minutes to complete. If you could highlight the differences you have observed between the two lots that would be very helpful.
Once I have receive the completed questionnaire, I will look at the protocol and see if there are any suggestions I can make that may improve the results. This information will also allow us to investigate this case internally and initiate additional testing where necessary. If the product was purchased in the last six months and is being used according to our Abpromise, we would be happy to replace or refund the antibody.
I look forward to receiving your reply.

Read More

Answer

Thank you for contacting Abcam.

I am sorry about the issues you have been having with ab94580 in western blot. The antibody is covered under our Abpromise for six months and is guaranteed to work in western blot on mouse & humansamples. If we cannot resolve the issue you are having with the antibody then I would be happy to either send a replacement antibody or to process a refund.

To be able to help you further in this matter, would you be able to answer some protocol questions:

1 - What sample type are you using?

2 - Are you seeing any bands, no bands etc?

3 - How much sample are you loading?

4 - What type of blocking agent are you using?

5 - What primary antibody concentrations have you tried and how long do you incubate for?

We do take all complaints on products seriously and we do our best to solve these problems.

I look forward to your reply.

Read More

Answer

Thank you very much for your reply.

That will not be a problem. This credit will not be refunded unlessyou contact ouraccounting department to request the refund,so the credit can be kept on your account andredeemed against the invoice of a future order. Please make sure your purchasingdepartment is aware of the credit and if they have any questions about how to use it, they can contact our accounting department at mailto:us.credits@abcam.com.

Please let me know if you have any questions or ifthere is anything else that we can do for you, and I'll be happy to help.

Read More

Question

Hi,

I ordered two TET2 antibody from Abcam in June and tested them on our inducible system, hoping that I may use them to detect endogenous TET2 protein. To validate that these antibodies work as described, I tested these antibodies, along with a positive control, and anti-TET2 antibodies purchased from other companies.

The TET2 overexpression cell line that we have generated express a Flag-tagged full-length human TET2. Without inducer, the exogenous TET2 levels are low; upon adding inducer, the TET2 levels increased over 10-fold. This was validated using anti-Flag antibody and thus Flag antibody was used as a positive control for the test.

Whole cell lysate (50 ug) from either un-treated or treated samples were loaded on SDS gel (Bio-Rad 4-20% TGX gel), transferred to PVDF membrane (Bio-Rad tank transfer, 100 V, 90 min), blocked in 5% milk for 1 hour, and incubated with various primary antibodies for overnight at 4 degree. Membranes were further incubated with HRP-conjugated secondary antibody for 2 hours at room temperature and developed using ECL.

As you may see from the attached figure, Flag antibody detected a band at ˜250 KD only when cells were treated with inducer (lower bands are proteolysis products). However, abcam anti-TET2 antibodies (ab94580, ab128178) failed to detected this band under neither conditions. An anti-TET2 antibody from one competitor did not work either. In contrast, an anti-TET2 antibody from another competitor successfully detected this band, and multiple proteolysis products, under induced condition, but not un-induced condition. The pattern recognized by this antibody looks very similar to that of Flag antibody, suggesting it can specifically recognize Flag-tagged TET2.

This experiment has been done twice and the same results were obtained.

Based on this, we conclude that the TET2 antibodies we purchased from Abcam did not work. According to your “Abpromise guarantee”, I am requesting a refund, or a credit for this order. Thanks.

Read More
Answer

Thank you for contacting us and letting us know about the trouble with these antibodies.

Thank you also for sending the information about your protocol and the image. Both of these antibodies may need to be used at lower dilutions than 1:2000 (ab128178 up to 1:500 and ab94580 at 1:1000), so this may be the reason there are no bands in the positive controls. I appreciate the time that you have spent working with these antibodies, so I have gone ahead and arranged the credit for you.

Your credit note ID is ***.

If payment has already been made on the original order and you wish to receive a refund, please ask your purchasing department to contact our accounting department at mailto:us.credits@abcam.com so that we may gather the correct information needed for the refund. To avoid confusion, please ensure your accounts department is aware of how the credit note is being used.

I hope this experience will not prevent you from purchasing other products from us in the future. Our Scientific Support team is always at your service, should you require further expert advice.

Read More

1-10 of 18 Abreviews or Q&A

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