Product Datasheet. Bromodeoxyuridine/BrdU Antibody (BU1/75 (ICR1)) NB Unit Size: 0.1 mg

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1 Product Datasheet Bromodeoxyuridine/BrdU Antibody (BU1/75 (ICR1)) NB Unit Size: 0.1 mg Store at 4C short term. Aliquot and store at -20C long term. Avoid freeze-thaw cycles. Reviews: 6 Publications: 89 Protocols, Publications, Related Products, Reviews, Research Tools and Images at: Updated 5/28/2018 v.20.1 Earn rewards for product reviews and publications. Submit a publication at Submit a review at

2 NB Bromodeoxyuridine/BrdU Antibody (BU1/75 (ICR1)) Product Information Unit Size Concentration Storage Clonality Clone Preservative Isotype Purity Buffer Product Description Host Species 0.1 mg 1.0 mg/ml Store at 4C short term. Aliquot and store at -20C long term. Avoid freeze-thaw cycles. Monoclonal BU1/75 (ICR1) 0.09% Sodium Azide IgG2a Protein G purified PBS Rat Human, Mouse, Monkey, Non-species specific Reactivity Notes Chemical. Monkey reactivity reported in scientific literature (PMID: ). Marker Specificity/Sensitivity Immunogen Product Application Details Applications Proliferation Marker This reacts with BrdU in single stranded DNA, BrdU attached to a protein carrier or free BrdU. It detects nucleated cells in S-Phase which have had BrdU incorporated into their DNA. Also reacts weakly with chlorodeoxyuridine, but does not cross react with thymidine or iododeoxyuridine. Made to Chemical BrdU Western Blot, Flow Cytometry, Immunocytochemistry/Immunofluorescence, Immunohistochemistry, Immunohistochemistry-Frozen, Immunohistochemistry- Paraffin, Immunocytochemistry Recommended Dilutions Western Blot, Flow Cytometry 1:25-1:200, Immunohistochemistry 1:10-1:500, Immunocytochemistry/Immunofluorescence 1:10-1:500, Immunohistochemistry- Paraffin 1:25-1:200, Immunohistochemistry-Frozen 1:10-1:500, Immunocytochemistry 1:200 Application Notes Page 1 of 6 v.20.1 Updated 5/28/2018 Clone BU1/75 (ICR1) can be used for labeling paraffin-embedded tissue sections fixed in formalin. Denaturation of the DNA is critical for successful staining of BrdU. This can be achieved by exposing cells to heat, or acid. For heat-induced epitope retrieval, 10mM citrate buffer ph6.0 is recommended. Alternatively, a 30 min incubation in 2M HCl can be performed. The HCl must then be neutralized for 2 min with 0.1 M Na2B4O7. Pretreatment of tissues with proteinase K should be avoided. Immunohistochemistry-Frozen was reported in scientific literature. Use in Western blot reported in scientific literature (PMID ).

3 Images Immunocytochemistry: Bromodeoxyuridine/BrdU Antibody (BU1/75 (ICR1)) [NB ] - Red fibers in the image denotes CldU incorporated DNA fibers that were probed with primary antibody diluted 1:200 in blocking buffe. This product works great for visualizing incorporated CldU into the DNA of proliferating cancer cells. This image was submitted via customer Review. Page 2 of 6 v.20.1 Updated 5/28/2018 Immunohistochemistry-Paraffin: Bromodeoxyuridine/BrdU Antibody (BU1/75 (ICR1)) [NB ] - BrdU-treated mouse duodenum. Image from verified customer review. Flow Cytometry: Bromodeoxyuridine/BrdU Antibody (BU1/75 (ICR1)) [NB ] - Analysis using the HRP conjugate of NB Staining of BrdU in cells using Rat anti BrdU:FITC (NB H) counterstained with propidium iodide. Immunocytochemistry/Immunofluorescence: Bromodeoxyuridine/BrdU Antibody (BU1/75 (ICR1)) [NB ] - Staining of mouse brain tissue, fixed with 4% PFA and dehydrated with 30% sucrose 30%. Image provided via product review by verified customer.

4 Immunohistochemistry-Paraffin: Bromodeoxyuridine/BrdU Antibody (BU1/75 (ICR1)) [NB ] - Mouse colon. Page 3 of 6 v.20.1 Updated 5/28/2018 Immunohistochemistry-Paraffin: Bromodeoxyuridine/BrdU Antibody (BU1/75 (ICR1)) [NB ] - Mouse colon. Immunohistochemistry-Paraffin: Bromodeoxyuridine/BrdU Antibody (BU1/75 (ICR1)) [NB ] - Mouse colon. Flow Cytometry: Bromodeoxyuridine/BrdU Antibody (BU1/75 (ICR1)) [NB ] - HL-60 cells were pulse labeled with BrdU for 45 minutes prior to harvesting and then incubated with primary antibody Rat anti BrdU clone BU1/75 diluted 1/100 followed by Rabbit anti Rat IgG secondary antibody (STAR17B) FITC-conjugated, diluted 1/200

5 Publications Al Mamun A, Yu H, Romana S, Liu F. Inflammatory Responses are Sex Specific in Chronic Hypoxic-Ischemic Encephalopathy. Cell Transplant Jan :00AM [PMID: ] (Mouse) Nam MO, Hahn S, Jee JH, Hwang TS. Effects of a small molecule R-spondin-1 substitute RS on a mouse intestinal organoid culture. Oncotarget Jan 19 [PMID: ] (Mouse) Kuroda MJ, Sugimoto C, Cai Y, Merino KM. High Turnover of Tissue Macrophages Contributes to Tuberculosis Reactivation in Simian Immunodeficiency Virus-Infected Rhesus Macaques. J. Infect. Dis Feb 08 [PMID: ] (Monkey) Sugimoto C, Merino KM, Hasegawa A, Wang X. Critical Role for Monocytes/Macrophages in Rapid Progression to AIDS in Pediatric Simian Immunodeficiency Virus-Infected Rhesus Macaques. J. Virol Sep 01 [PMID: ] (IHC, Monkey) Kottemann MC, Conti BA, Lach FP, Smogorzewska A. Removal of RTF2 from Stalled Replisomes Promotes Maintenance of Genome Integrity. Mol. Cell 2017 Dec 15 [PMID: ] Apostolopoulou M, Kiehl TR, Winter M et al. Non-monotonic Changes in Progenitor Cell Behavior and Gene Expression during Aging of the Adult V-SVZ Neural Stem Cell Niche Stem Cell Reports Nov 08 [PMID: ] (IHC-Fr, Mouse) Tang JJ, Podratz JL, Lange M et al. Mechano growth factor, a splice variant of IGF-1, promotes neurogenesis in the aging mouse brain Mol Brain 2017 Jul 07 [PMID: ] (IHC-P, Mouse) S B, K S, S A et al. RAD51 interconnects between DNA replication, DNA repair and immunity. NCBI-GEO datasets. Feb :00AM [PMID: ] (IHC, Human) Guo K, Li CH, Wang XY et al. Germ stem cells are active in postnatal mouse ovary under physiological conditions. Mol. Hum. Reprod. May :00AM [PMID: ] (ICC/IF, Mouse) Garofalo S, Grimaldi A, Chece G et al. The glycoside oleandrin reduces glioma growth with direct and indirect effects on tumor cells. J. Neurosci. Mar :00AM [PMID: ] (IHC, Mouse) Huang HM, Bauer DC, Lelliott PM et al. A novel ENU-induced ankyrin-1 mutation impairs parasite invasion and increases erythrocyte clearance during malaria infection in mice. Sci Rep Nov 16 [PMID: ] (IHC, Mouse) Details: This citation used the Biotin version of this antibody. Seidel P, Remus M, Delacher M et al. Epidermal Nbn deletion causes premature hair loss and a phenotype resembling psoriasiform dermatitis. Oncotarget 2016 Apr 26 [PMID: ] (IHC) More publications at Page 4 of 6 v.20.1 Updated 5/28/2018

6 Procedures Flow Cytometry Protocol for BrdU Antibody (NB ) Flow Cytometry Analysis: Page 5 of 6 v.20.1 Updated 5/28/2018 Prepare the following solutions before proceeding: Phosphate buffered saline (PBS) 2N HCl, 0.5% Triton X-100 PBS containing 0.05% Tween-20 PBS containing 1% BSA (PBS/BSA) 10mg/ml Propidium iodide (PI) As a positive control, BrdU labeled cells maybe obtained from Phoenix Flow Systems ( catalogue number ACNC Add BrdU to the cell suspension in culture medium to a final concentration of 10 umol/l and incubate for 30 minutes in a CO2 incubator at 37C. 2. Wash cells twice with PBS/BSA, and resuspend in PBS 3. Add cells slowly into 5ml of 70% ethanol at -20C, mixing continuously (vortex preferred). Incubate on ice for 30 minutes. 4. Centrifuge at 500g for 10 minutes, decant supernatant, and resuspend cell pellet. 5. Add 2ml 2N HCl, 0.5% Triton X-100 and incubate the cells for 30 minutes at room temperature (preferably on a rocking platform). 6. Centrifuge at 500g for 10 minutes, decant supernatant and resuspend in 3 ml 0.1M Na2B4O7, ph Centrifuge at 500g for 10 minutes, decant supernatant and resuspend the cells in PBS/BSA % Tween 20. Adjust cell concentration to 1 x 10(7)/ml 8. Aliquot 100ul of cell suspension into required number of 12 x 75mm tubes. 9. Incubate the cells with the anti-brdu antibody at the recommended dilution for 30 minutes at room temperature. 10. Add 2 mls PBS/BSA and centrifuge the cells at 1000rpm for 5 minutes. 11. If a secondary antibody layer is required then decant the wash and incubate the cells with the secondary antibody for 30 minutes at room temperature. If no secondary antibody layer is required then proceed to step Wash the cells after the secondary antibody layer by repeating step Decant off the wash and add 1ml PBS containing 10ug/ml PI (Dilute the 10mg/ml solution of PI 1/1000 in a suitable volume of PBS) 14. Analyse cells by flow cytometry following the manufacturer's instructions. The PI should be read on the appropriate channel set to the Peak/Area and not log scale. IMMUNOHISTOLOGY Formalin-fixed paraffin-embedded tissue sections: Clone BU1/75 (ICR1) can be used for labelling paraffin-embedded tissue sections fixed in formalin. Pretreatment of tissues with heat-induced epitope retrieval using 10mM citrate buffer ph6.0 is recommended. Pretreatment of tissues with proteinase K should be avoided.

7 Novus Biologicals USA 8100 Southpark Way, A-8 Littleton, 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 HAF005 Goat anti-rat IgG Secondary Antibody [HRP (Horseradish Peroxidase)] NB7115 Goat anti-rat IgG (H+L) Secondary Antibody [HRP] NBP Rat IgG2a Isotype Control (KLH/G2a-1-1) NB B Bromodeoxyuridine/BrdU Antibody (BU1/75 (ICR1)) [Biotin] 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: