Product Datasheet. TRF-2 Antibody NB Unit Size: 0.1 ml

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1 Product Datasheet TRF-2 Antibody NB Unit Size: 0.1 ml Store at 4C short term. Aliquot and store at -20C long term. Avoid freeze-thaw cycles. Reviews: 1 Publications: 54 Protocols, Publications, Related Products, Reviews, Research Tools and Images at: Updated 2/20/2019 v.20.1 Earn rewards for product reviews and publications. Submit a publication at Submit a review at

2 NB TRF-2 Antibody Product Information Unit Size Concentration Storage Clonality Preservative Isotype Purity Buffer Target Molecular Weight 0.1 ml 1.0 mg/ml Store at 4C short term. Aliquot and store at -20C long term. Avoid freeze-thaw cycles. Polyclonal 0.02% Sodium Azide IgG Immunogen affinity purified PBS 56 kda Page 1 of 6 v.20.1 Updated 2/20/2019 Product Description Host Rabbit Gene ID 7014 Gene Symbol TERF2 Species Human, Mouse, Rat, Chinese Hamster, Primate Marker Telomeres marker Immunogen Baculovirus purified TRF2 protein. Product Application Details Applications Recommended Dilutions Application Notes Western Blot, Simple Western, Chromatin Immunoprecipitation, Immunocytochemistry/Immunofluorescence, Immunohistochemistry, Immunohistochemistry-Paraffin, Immunoprecipitation Western Blot 1:2000-1:5000, Simple Western 1:25, Chromatin Immunoprecipitation 1:10-1:500, Immunohistochemistry 1:200-1:500, Immunocytochemistry/Immunofluorescence 1:50-1:200, Immunoprecipitation, Immunohistochemistry-Paraffin 1:200-1:500 This TRF2 antibody is useful for ChIP, Immunocytochemistry/Immunofluorescence, Immunohistochemistry paraffin embedded sections and Western blot, where a band at approx. 56 kda is seen. Immunoprecipitation was reported in scientific literature. In ICC/IF, nuclear staining was observed in HeLa cells. In IHC, nuclear staining was observed in xenografted human breast cancer tissue. The observed molecular weight of the protein may vary from the listed predicted molecular weight due to post translational modifications, post translation cleavages, relative charges, and other experimental factors. Prior to immunostaining paraffin tissues, antigen retrieval with sodium citrate buffer (ph 6.0) is recommended. In Simple Western only ul of the recommended dilution is used per data point. Separated by Size-Wes, Sally Sue/Peggy Sue. The observed molecular weight of the protein may vary from the listed predicted molecular weight due to post translational modifications, post translation cleavages, relative charges, and other experimental factors.

3 Images Immunocytochemistry/Immunofluorescence: TRF-2 Antibody [NB ] - HeLa cells were fixed for 10 minutes using 10% formalin and then permeabilized for 5 minutes using 1X TBS + 0.5% Triton X-100. The cells were incubated with antibody at a 1:200 dilution overnight at 4 degrees Celsius and detected with DyLight 488 (Green) at a 1:500 dilution. Alpha tubulin was used as a co-stain at a 1:1000 dilution and detected with Dylight 550 (Red). Nuclei were detected with DAPI (Blue) at 2.0 ug/ml in 1X PBS. Cells were imaged using a 40X objective. Page 2 of 6 v.20.1 Updated 2/20/2019 Simple Western: TRF-2 Antibody [NB ] - Lane view shows a specific band for TRF2 in 1.0 mg/ml of HeLa lysate. This experiment was performed under reducing conditions using the kda separation system. Western Blot: TRF-2 Antibody [NB ] - Analysis of HeLa whole cell lysate (A), HeLa nuclear cell lysate (B), k562 cell lysate (C), HepG2 cell lysate (D), NIH/3T3 cell lysate (E), CHO cell lysate (F), PC12 cell lysate (G), and Cos7 cell lysate (H) using antibody at a concentration of 2 ug/ml. Immunohistochemistry-Paraffin: TRF-2 Antibody [NB ] - Analysis in xenografted human breast cancer tissue using DAB with hematoxylin counterstain.

4 Immunohistochemistry-Paraffin: TRF-2 Antibody [NB ] - Analysis of FFPE human breast cancer tissue with rabbit polyclonal TRF2 antibody at a dilution of 1:200. The staining was developed with HRP-DAB detection method and the counterstaining was performed using hematoxylin. This TRF2 antibody generated an expected nuclear signal in all the cancer cells and the stromal cells. In the tested section, only a subset of myoepithelial cells showed positivity for this protein. Page 3 of 6 v.20.1 Updated 2/20/2019 Chromatin Immunoprecipitation: TRF-2 Antibody [NB ] - Analysis in mouse. Titrated TRF2 antibody to determine concentration required for ChIP experiment. Image from verified customer review. Immunohistochemistry-Paraffin: TRF-2 Antibody [NB ] - Analysis of FFPE human breast cancer tissue with rabbit polyclonal TRF2 antibody at 1:200 dilution. The staining was developed with HRP- DAB detection method and the counterstaining was performed using hematoxylin. This TRF2 antibody generated an expected nuclear signal in all the cancer cells and the stromal cells. In the tested section, only a subset of myoepithelial cells showed positivity for this protein.

5 Publications Mukherjee AK, Sharma S, Sengupta S et al. Telomere length-dependent transcription and epigenetic modifications in promoters remote from telomere ends. PLoS Genet. Nov :00AM [PMID: ] (ChIP, Human) Chow TT, Shi X, Wei JH et al. Local enrichment of HP1alpha at telomeres alters their structure and regulation of telomere protection. Nat Commun Sep :00AM [PMID: ] (ChIP, Human) Ilic Aleksandar, Lu Sumin, Bhatia Vikram et al. Ubiquitin C-terminal hydrolase isozyme L1 is associated with shelterin complex at interstitial telomeric sites. Epigenetics & Chromatin 2017 [PMID: ] (IP, Human) Gu P, Jia S, Takasugi T et al. CTC1-STN1 coordinates G- and C-strand synthesis to regulate telomere length Aging Cell May :00AM [PMID: ] (Human) Nelson ND, Dodson LM, Escudero L et al. The C-terminal extension unique to the long isoform of the shelterin component TIN2 enhances its interaction with TRF2 in a phosphorylation- and dyskeratosis congenita-clusterdependent fashion. Mol. Cell. Biol. Mar :00AM [PMID: ] (IP) Li JS, Miralles Fuste J, Simavorian T, Bartocci C. TZAP: A telomere-associated protein involved in telomere length control. Science Feb 10 [PMID: ] (ICC/IF, Human) Biswas U, Stevense M, Jessberger R. SMC1a Substitutes for Many Meiotic Functions of SMC1b but Cannot Protect Telomeres from Damage Curr. Biol Jan 22 [PMID: ] (ICC/IF) Zhang J, Tu Z, Watanabe Y, Shibuya H. Distinct TERB1 Domains Regulate Different Protein Interactions in Meiotic Telomere Movement Cell Rep Nov 14 [PMID: ] (ICC/IF, Mouse) Benhamou Y, Picco V, Raybaud H et al. Telomeric repeat-binding factor 2: a marker for survival and anti-egfr efficacy in oral carcinoma Oncotarget Jul :00AM [PMID: ] (WB, Human) Tu Z, Bayazit MB, Liu H et al. Speedy A-Cdk2 binding mediates initial telomere-nuclear envelope attachment during meiotic prophase I independent of Cdk2 activation. Proc. Natl. Acad. Sci. U.S.A. Dec :00AM [PMID: ] (WB, Mouse) Knecht H, Mai S. The Use of 3D Telomere FISH for the Characterization of the Nuclear Architecture in EBV-Positive Hodgkin's Lymphoma. Methods Mol. Biol. Nov :00AM [PMID: ] Sishc BJ, Nelson CB, McKenna MJ et al. Telomeres and Telomerase in the Radiation Response: Implications for Instability, Reprograming, and Carcinogenesis. Front Oncol 2015 Nov 24 [PMID: ] More publications at Page 4 of 6 v.20.1 Updated 2/20/2019

6 Page 5 of 6 v.20.1 Updated 2/20/2019 Procedures Western Blot protocol for TRF2 Antibody (NB ) 1. Perform SDS-PAGE on samples to be analyzed, loading 40 ug of total protein per lane. 2. Transfer proteins to membrane according to the instructions provided by the manufacturer of the membrane and transfer apparatus. 3. Stain according to standard Ponceau S procedure (or similar product) to assess transfer success, and mark molecular weight standards where appropriate. 4. Rinse the blot. 5. Block the membrane using standard blocking buffer for at least 1 hour. 6. Wash the membrane in wash buffer three times for 10 minutes each. 7. Dilute primary antibody in blocking buffer and incubate 1 hour at room temperature. 8. Wash the membrane in wash buffer three times for 10 minutes each. 9. Apply the diluted HRP conjugated secondary antibody in blocking buffer (as per manufacturers instructions) and incubate 1 hour at room temperature. 10. Wash the blot in wash buffer three times for 10 minutes each (this step can be repeated as required to reduce background). 11. Apply the detection reagent of choice in accordance with the manufacturers instructions. Note: Tween-20 can be added to the blocking or antibody dilution buffer at a final concentration of %. *The above information is only intended as a guide. The researcher should determine what protocol best meets their needs. Please follow safe laboratory procedures. Immunocytochemistry/Immunofluorescence Protocol for TRF2 Antibody (NB ) Culture cells to appropriate density in 35 mm culture dishes or 6-well plates. 1. Remove culture medium and add 10% formalin to the dish. Fix at room temperature for 30 minutes. 2. Remove the formalin and add ice cold methanol. Incubate for 5-10 minutes. 3. Remove methanol and add washing solution (i.e. PBS). Be sure to not let the specimen dry out. Wash three times for 10 minutes. 4. To block nonspecific antibody binding incubate in 10% normal goat serum from 1 hour to overnight at room temperature. 5. Add primary antibody at appropriate dilution and incubate at room temperature from 2 hours to overnight at room temperature. 6. Remove primary antibody and replace with washing solution. Wash three times for 10 minutes. 7. Add secondary antibody at appropriate dilution. Incubate for 1 hour at room temperature. 8. Remove antibody and replace with wash solution, then wash for 10 minutes. Add Hoechst to wash solution at 1:25,0000 and incubate for 10 minutes. Wash a third time for 10 minutes. 9. Cells can be viewed directly after washing. The plates can also be stored in PBS containing Azide covered in Parafilm (TM). Cells can also be cover-slipped using Fluoromount, with appropriate sealing. *The above information is only intended as a guide. The researcher should determine what protocol best meets their needs. Please follow safe laboratory procedures.

7 Novus Biologicals USA E. Briarwood Avenue Centennial, CO USA Phone: Toll Free: Fax: Novus Biologicals Canada 461 North Service Road West, Unit B37 Oakville, ON L6M 2V5 Canada Phone: Toll Free: Fax: Novus Biologicals Europe 19 Barton Lane Abingdon Science Park Abingdon, OX14 3NB, United Kingdom Phone: (44) (0) Free Phone: Fax: (44) (0) General Contact Information Technical Support: Orders: General: Products Related to NB NB800-PC1 HAF008 NB7160 NBP HeLa Whole Cell Lysate Goat anti-rabbit IgG Secondary Antibody [HRP (Horseradish Peroxidase)] Goat anti-rabbit IgG (H+L) Secondary Antibody [HRP] Rabbit IgG Isotype Control Limitations This product is for research use only and is not approved for use in humans or in clinical diagnosis. Primary Antibodies are guaranteed for 1 year from date of receipt. For more information on our 100% guarantee, please visit Earn gift cards/discounts by submitting a review: Earn gift cards/discounts by submitting a publication using this product: