Bromodeoxyuridine/BrdU Antibody (BU1/75 (ICR1))

Images

 
Immunocytochemistry: Bromodeoxyuridine/BrdU Antibody (BU1/75 (ICR1)) [NB500-169] - Red fibers in the image denotes CldU incorporated DNA fibers that were probed with primary antibody diluted 1:200 in blocking buffe. ...read more
Immunohistochemistry-Paraffin: Bromodeoxyuridine/BrdU Antibody (BU1/75 (ICR1)) [NB500-169] - BrdU-treated mouse duodenum. Image from verified customer review.
Flow Cytometry: Bromodeoxyuridine/BrdU Antibody (BU1/75 (ICR1)) [NB500-169] - Analysis using the HRP conjugate of NB500-169. Staining of BrdU in cells using Rat anti BrdU:FITC (NB500-169H) counterstained with propidium ...read more
Immunocytochemistry/ Immunofluorescence: Bromodeoxyuridine/BrdU Antibody (BU1/75 (ICR1)) [NB500-169] - Staining of mouse brain tissue, fixed with 4% PFA and dehydrated with 30% sucrose 30%. Image provided via product ...read more
Immunohistochemistry-Paraffin: Bromodeoxyuridine/BrdU Antibody (BU1/75 (ICR1)) [NB500-169] - Mouse colon.
Immunohistochemistry-Paraffin: Bromodeoxyuridine/BrdU Antibody (BU1/75 (ICR1)) [NB500-169] - Mouse colon.
Immunohistochemistry-Paraffin: Bromodeoxyuridine/BrdU Antibody (BU1/75 (ICR1)) [NB500-169] - Mouse colon.
Flow Cytometry: Bromodeoxyuridine/BrdU Antibody (BU1/75 (ICR1)) [NB500-169] - HL-60 cells were pulse labeled with BrdU for 45 minutes prior to harvesting and then incubated with primary antibody Rat anti BrdU clone ...read more

Product Details

Summary
Reactivity Hu, Mu, Mk, NASpecies Glossary
Applications WB, Flow, ICC/IF, IHC, IHC-Fr, IHC-P, ICC/IF
Clone
BU1/75 (ICR1)
Clonality
Monoclonal
Host
Rat
Conjugate
Unconjugated
Concentration
1.0 mg/ml

Bromodeoxyuridine/BrdU Antibody (BU1/75 (ICR1)) Summary

Immunogen
Made to Chemical BrdU
Localization
Nuclear
Marker
Proliferation Marker
Specificity
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.
Isotype
IgG2a
Clonality
Monoclonal
Host
Rat
Purity
Protein G purified
Innovator's Reward
Test in a species/application not listed above to receive a full credit towards a future purchase.

Applications/Dilutions

Dilutions
  • Western Blot
  • Flow Cytometry 1:25-1:200
  • Immunocytochemistry/Immunofluorescence 1:10-1:500
  • Immunohistochemistry 1:10-1:500
  • Immunohistochemistry-Frozen 1:10-1:500
  • Immunohistochemistry-Paraffin 1:25-1:200
  • Immunocytochemistry 1:200
Application Notes
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 27425845).
Reviewed Applications
Read 6 Reviews rated 4.7
using
NB500-169 in the following applications:

Publications
Read Publications using
NB500-169 in the following applications:

Reactivity Notes

Chemical. Monkey reactivity reported in scientific literature (PMID: 28566378).

Packaging, Storage & Formulations

Storage
Store at 4C short term. Aliquot and store at -20C long term. Avoid freeze-thaw cycles.
Buffer
PBS
Preservative
0.09% Sodium Azide
Concentration
1.0 mg/ml
Purity
Protein G purified

Alternate Names for Bromodeoxyuridine/BrdU Antibody (BU1/75 (ICR1))

  • 5-bromo-2-deoxyuridine
  • BrdU
  • Bromodeoxyuridine
  • BUdr

Background

The immunocytochemical detection of bromodeoxyuridine (BrdU) incorporated into DNA is a powerful tool to study the cytokinetics of normal and neoplastic cells. In vitro or in vivo labeling of cells with the thymidine analogue BrdU and the subsequent detection of incorporated BrdU with specific anti-BrdU monoclonal antibodies is an accurate and comprehensive method to quantitate the degree of DNA-synthesis. BrdU is incorporated into the newly synthezised DNA of the S-phase cells and can thus provide an estimate for the fraction of cells in S-phase. Also dynamic proliferative information (such as the S-phase transit rate and the potential doubling time) can be obtained, by means of bivariate BrdU/DNA flow cytometric analysis.

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.

Publications for Bromodeoxyuridine/BrdU Antibody (NB500-169)(89)

We have publications tested in 5 confirmed species: Human, Mouse, Rat, Monkey, drosophila.

We have publications tested in 7 applications: FLOW, ICC/IF, IHC, IHC-Fr, IHC-FrFl, IHC-P, WB.


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FLOW
(1)
ICC/IF
(26)
IHC
(26)
IHC-Fr
(12)
IHC-FrFl
(1)
IHC-P
(6)
WB
(2)
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Filter By Species
Human
(6)
Mouse
(26)
Rat
(3)
Monkey
(2)
drosophila
(1)
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Showing Publications 1 - 10 of 89. Show All 89 Publications.
Publications using NB500-169 Applications Species
Al Mamun A, Yu H, Romana S, Liu F. Inflammatory Responses are Sex Specific in Chronic Hypoxic-Ischemic Encephalopathy. Cell Transplant Jan 1 2018 [PMID: 29692197] (Mouse) Mouse
Padilla SL. Growing Up POMC: Pro-opiomelanocortin in the Developing Brain. Columbia University. 2011
Nam MO, Hahn S, Jee JH, Hwang TS. Effects of a small molecule R-spondin-1 substitute RS-246204 on a mouse intestinal organoid culture. Oncotarget. 2018 Jan 19 [PMID: 29464078] (Mouse) 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. 2018 Feb 08 [PMID: 29432596] (Monkey) 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: 29290612]
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. 2017 Nov 08 [PMID: 29129683] (IHC-Fr, Mouse) 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: 28683812] (IHC-P, Mouse) IHC-P Mouse
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. 2017 Sep 01 [PMID: 28566378] (IHC, Monkey) IHC Monkey
S B, K S, S A et al. RAD51 interconnects between DNA replication, DNA repair and immunity. NCBI-GEO dataSets. Feb 18 2017 [PMID: 28334891] (IHC, Human) IHC Human
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 14 2017 [PMID: 28292827] (IHC, Mouse) IHC Mouse
Show All 89 Publications.

Reviews for Bromodeoxyuridine/BrdU Antibody (NB500-169) (6) 4.76

Average Rating: 4.7
(Based on 6 reviews)
We have 6 reviews tested in 2 species: Human, Mouse.

Reviews using NB500-169:
Filter by Applications
ICC
(1)
IHC
(2)
IHC-P
(2)
IHC-Fr
(1)
All Applications
Filter by Species
Human
(1)
Mouse
(5)
All Species
Images Ratings Applications Species Date Details
Immunocytochemistry Bromodeoxyuridine/BrdU NB500-169
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reviewed by:
Anonymous
ICC Human 06/07/2017
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Summary

ApplicationImmunocytochemistry
Sample TestedDAN fiber spreads of head and neck cancer cells
SpeciesHuman
Lot1015

Comments

CommentsComments: We routinely use product # NB500-169 in performing DNA fiber assay. This product works great for visualizing incorporated CldU into the DNA of proliferating cancer cells. This product has a high signal to noise ratio. Overall, we rate this product highly for immunofluorescent detection of ClDU in DNA fiber assay.
Blocking details: 10% Normal goat serum + 0.1% Triton X-100 in PBS for 1hr at room temperature
Primary Antibody (# NB500-169): 1:200 in blocking buffer for 1:30 hrs in humidified chamber
Secondary Antibody: Alexa fluor 555 goat anti-rat, 1:300 in blocking buffer for 1hr in humidified chamber
Detection: Fluorescent microscopy
Fixation and Denaturation details: Cells are lysed in lysis buffer (1% SDSA, 50 mM EDTA, 200 mM Tris-Cl), and spotted on glass slide. Glass slide is tilted to enable DAN spreading. Upon air drying, DNA is fixed in methanol:acetonbe ( 3:1) for 15 min and then denatured in 2.5 N HCl, then neutralized in PBS (pH 8).
 
reviewed by:
Parth Patel
IHC Mouse 04/27/2017
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Summary

ApplicationImmunohistochemistry
Sample TestedMouse Pancreas
SpeciesMouse
 
reviewed by:
Parth Patel
IHC Mouse 04/18/2017
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Summary

ApplicationImmunohistochemistry
Sample TestedMouse Pancreas,mouse liver,Mouse Spleen
SpeciesMouse
Immunohistochemistry-Paraffin Bromodeoxyuridine/BrdU NB500-169
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reviewed by:
Anonymous
IHC-P Mouse 05/29/2014
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Summary

ApplicationImmunohistochemistry-Paraffin
Sample TestedBrdU-treated mouse duodenum
SpeciesMouse
CommentsBeautiful antibody. Worked perfectly at every concentration and every epitope retrieval we tried.

Primary Anitbody

Dilution Ratio1:1500

Details

Fixation DetailsFFPE HIER

Comments

CommentsBeautiful antibody. Worked perfectly at every concentration and every epitope retrieval we tried.
Immunohistochemistry-Frozen Bromodeoxyuridine/BrdU NB500-169
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reviewed by:
Valeria Bortolotto
IHC-Fr Mouse 08/07/2013
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Summary

ApplicationImmunohistochemistry-Frozen
Sample Testedmouse brain
SpeciesMouse
Lot1210109
CommentsFor many years our laboratory has used the antibody NB500-169 especially for immunohistochemistry on frozen tissue. Recently we also tried the same antibody on cells and the result was excellent, no aspecific signal, no background and because of this it is easy to detect with confocal and fluorescente microscope especially if it is associated with double or triple staining.

Blocking

Blocking DetailsNormal Goat Serum (NGS) 10% + Triton X-100 0.5% in TBS 1 h at 4°C

Primary Anitbody

Dilution Ratio1:200; NGS 2% + Triton X-100 0.1% in TBS O/N at 4°C

Secondary Antibody

Secondary DescriptionGoat anti-Rat IgG Cy5
Secondary Manufacturer Cat#Jackson Immunofluorescence 112-175-143
Secondary Concentration1:300

Details

Detection NotesConfocal microscope
Fixation DetailsFixation: PFA 4%, Antigen Retrieval: HCl 2N 30 min at 37°C
Wash DescriptionTBS + Tween 0.1% 3 times for 5 min at RT

Comments

CommentsFor many years our laboratory has used the antibody NB500-169 especially for immunohistochemistry on frozen tissue. Recently we also tried the same antibody on cells and the result was excellent, no aspecific signal, no background and because of this it is easy to detect with confocal and fluorescente microscope especially if it is associated with double or triple staining.
 
reviewed by:
Donald Maris
IHC-P Mouse 04/14/2010
View

Summary

ApplicationImmunohistochemistry-Paraffin
Sample Testedmouse brain
SpeciesMouse
LotIMAS252072
CommentsAn overnight incubation of the primary antibody at 1:300 dilution produced the most optimal labeling at 4 degrees.
Pub Med ID19960516
FileView PDF

Blocking

Blocking DetailsBlocking Buffer: 5% normal goat serum, Blocking Time: 1 hour, Blocking Temp: room temperature

Primary Anitbody

Dilution RatioDilution Ratio: 1:300, Incubation Dilution Buffer: 5% norm goat serum,.6% triton-x in 0.01M PBS, Incubation Time:24hr, Temp:4C

Secondary Antibody

Secondary DescriptionSecondary Ab: Alexa fluor 647:, Secondary Ab Dilution: 1:400
Secondary Manufacturer Cat#A21247

Details

Detection NotesDetection Method: Fluorescence, Negative Control: No primary, Antigen Retrieval: none required
Fixation DetailsSample Preparation: perfused

Comments

CommentsAn overnight incubation of the primary antibody at 1:300 dilution produced the most optimal labeling at 4 degrees.

Product General Protocols

View specific protocols for Bromodeoxyuridine/BrdU Antibody (NB500-169): Find general support by application which include: protocols, troubleshooting, illustrated assays, videos and webinars.

Video Protocols

WB Video Protocol
ICC/IF Video Protocol

FAQs for Bromodeoxyuridine/BrdU Antibody (NB500-169). (Showing 1 - 2 of 2 FAQs).

  1. Is the NB500-169 still available in the concentration of 2mg/ml? We ordered five vials in the past month with the unit size 0.1 mg. Three of them arrived with a concentration of 2mg/ml, the last two vial with a concentration of 1mg/ml. Can we influence the concentration when ordering?
    • In terms of the concentration of the vials, unfortunately sometimes the concentrations of our products may change. (The concentration of each vial is that stated on the label.) I am sorry, but it is not possible for you to order specifically 2 mg/ml vials of NB500-169. (If you require a concentration of 2 mg/ml, you may be interested in our Antibody Concentration and Clean Up Kit (product code for 1 spin column is 861-0005; 3 spin columns is 861-0010), which you could use to reduce the volume of the antibody by half and hence double the concentration.)
  2. I am planning on doing DNA fiber fish using: 5'-iodo-2-deoxyuridine (IdU) and 5'-chloro-2-deoxiuridine (CldU). What antibodies from Novus do you recommend?
    • Catalog number NB500-169 is your best candidate for IdU as it has been very well supported by 52 references. As far as antibodies for CIdU is concerned, unfortunately currently we do not carry any antibodies for it however because of the high similarity of CldU and IdU with BrdU, the anti-BrdU antibodies also react with these modified nucleosides and you can totally use NB500-169 to detect both IdU and CldU, the only problem is, this might complicate the multiple labeling of cells.

Secondary Antibodies

 

Isotype Controls

Other Available Formats

Alexa Fluor 405 Labeled NB500-169AF405
Alexa Fluor 488 Labeled NB500-169AF488
Alexa Fluor 647 Labeled NB500-169AF647
Alexa Fluor 700 Labeled NB500-169AF700
Biotin Labeled NB500-169B
DyLight 488 Labeled NB500-169G
DyLight 550 Labeled NB500-169R
DyLight 650 Labeled NB500-169C
FITC Labeled NB600-720

Additional Array Products

Blogs on Bromodeoxyuridine/BrdU.

BrDU (Bromodeoxyuridine)
The thymidine synthetic nucleoside analogue bromodeoxyuridine (BrdU) has a long, colorful history of repeated use in molecular and cytokinetic studies, as detailed in reviews by Vanderlaan and Dolbeare (1,2).  Because BrDU is only incorporated in...  Read full blog post.

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Recent Reviews

4.7
5
4
4
2
3
0
2
0
1
0

Anonymous
06/07/2017
Application: ICC
Species: Human

 
Parth Patel
04/27/2017
Application: IHC
Species: Mouse

 
Parth Patel
04/18/2017
Application: IHC
Species: Mouse