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由来生物
mouse
品質水準
抗体製品の状態
purified immunoglobulin
抗体製品タイプ
primary antibodies
クローン
131-14871, monoclonal
AH4H7-1, monoclonal
交差性（ホモロジーによる予測）
all
メーカー／製品名
Chemicon®
テクニック
flow cytometry: suitable
immunocytochemistry: suitable
immunohistochemistry: suitable
アイソタイプ
IgG1
輸送温度
wet ice
ターゲットの翻訳後修飾
unmodified
詳細
ブロモデオキシウリジン（BrdU）はチミジンアナログであり、DNA合成中にDNAに特異的に結合します。抗ブロモデオキシウリジンモノクローナル抗体を用いて、BrdUが取込まれた細胞を特定できます。 この免疫学的検出法には、複製中の細胞を特定するための放射性チミジン取り込み法と比較して、いくつかの利点があります。トリチウム標識細胞のオートラジオグラフィーに必要となる数日間の待機を行わずに、標識と検出を同日に実施可能であり、最適な曝露時間を得るために複数の検体を使用する必要がなくなります。さらに、フローサイトメトリー分析による抗ブロモデオキシウリジン染色により、複数のパラメータを同時に評価することができます。 抗ブロモデオキシウリジンモノクローナル抗体は、血中（Campana et al., 1988）、組織中（Schutte et al., 1987; Hayashi, et al., 1988）、腫瘍中（Hoshino et al., 1986; Morstyn et al., 1983）の増殖細胞を特定するために用いられるほか、形質細胞の標識指数を決定するためにも用いられています（Greipp et al., 1985）。
特異性
ブロモデオキシウリジンに結合し、ヨードウリジン（10%）と交差反応します。抗ブロモデオキシウリジン抗体は、フルオロデオキシウリジンや、チミジンやウリジンなどの内因性細胞成分と交差反応しません。
免疫原
ブロモデオキシウリジン-ウシ血清アルブミン濃縮液
アプリケーション
Immunohistochemistry: (6 μg/ml)
Flow Cytometry: (0.2 μg/100 μl/10E6 cells) Optimal working dilutions must be determined by end user.
APPLICATIONS
Flow cytometry:The method below is based on that of M. Vanderlaan et al. (1986). Variations of this method exist in the literature, one consideration being the effect various fixation procedures have on the light-scattering properties of different cell populations. Procedure:
1. To label cells, pulse with 10 μM bromodeoxyuridine for 30 minutes. Harvest cells from culture.
2. Fix cells in 70% ethanol at +2-8°C for at least 30 min. Extract histones by resuspending cells in 1 mL chilled 0.1 M HCI containing 0.5% Triton X-100; incubate the suspension on ice for 10 minutes. Dilute acid with 5 mL distilled water and centrifuge at 200 x g for 10 min. Resuspend cells in 2 mL distilled water.
3. Denature cellular DNA by submerging the cell suspension into a boiling water bath for 10 min. Afterwards, quickly cool by placing the cell suspension in an ice slurry for several minutes. Wash cells in PBS that contains 0.5% Triton X-100.
4. Resuspend the cells (1-2 x 10 6 cells) in 100 μL of solution containing approximately 2 μg/mL anti-bromodeoxyuridine antibody diluted in PBS containing 0.1% BSA (0.2 μg/test). Incubate for 30 min at room temperature. Wash cells with PBS.
5. Resuspend cells in 100 μL of diluted goat anti-mouse IgG-FlTC Wash cells with PBS.
APPLICATIONS (Cont.)
Immunohistochemistry: Below is a procedure for staining cells that have been labeled with BrdU in vivo or in vitro. The procedure is based on the methods of B. Schutte et al. (1987) and D. Campana et al. (1988).
Preparation of tissue:
Inject animal with 50 mg BrdU/kg body weight. Sacrifice animal one hour later and remove organ or tissue under study. Embed tissue in OCT medium and snap-freeze by immersion into liquid nitrogen.Cut 4 mm frozen sections with a cryostat. Place sections on either albumin- or gelatin-coated slides.
Preparation of cells:
Pulse cells with 10 mM BrdU for 60 min. Cells grown on coverslips, or cytocentrifuge preparations made from cells grown in suspension, can be used for anti-bromodeoxyuridine staining according to the procedure below.
Procedure
1. Fix tissue sections or cells (on slide or coverglass) by immersing in absolute methanol for 10 minutes at +2-8°C. Air dry after removing from fixative. The slides can be stored at -20°C in a sealed box, or rehydrated to prepare for the assay procedure. To rehydrate, immerse in PBS for 3 min.
2. Denature DNA by incubating the slides in 2 N HCI for 60 min at +37°C.
3. Neutralize the acid by immersing the slides in 0.1 M borate buffer, pH 8.5. Change the buffer twice over a 10 min period.
4. Wash slides with PBS, changing the solution three times over a 10 min period.
5. Place slides in a humidified chamber (e.g., a sealed plastic box layered with wet paper towels) and cover cells with 150-300 μL of solution containing approximately 6 μg/mL anti-bromodeoxyuridine antibody diluted in PBS with 0.1% BSA. Incubate for 60 min at room temperature.
6. Wash slides with PBS, changing the solution three times over a 10 min period.
7. Apply optimal dilution of a second antibody conjugate (e.g., anti-mouse IgG-peroxidase), incubate, wash, and perform detection with a substrate that produces an insoluble product. After detection, counterstain with Harris-modified hematoxylin if desired. Slides can then be dehydrated and mounted.
Flow Cytometry: (0.2 μg/100 μl/10E6 cells) Optimal working dilutions must be determined by end user.
APPLICATIONS
Flow cytometry:The method below is based on that of M. Vanderlaan et al. (1986). Variations of this method exist in the literature, one consideration being the effect various fixation procedures have on the light-scattering properties of different cell populations. Procedure:
1. To label cells, pulse with 10 μM bromodeoxyuridine for 30 minutes. Harvest cells from culture.
2. Fix cells in 70% ethanol at +2-8°C for at least 30 min. Extract histones by resuspending cells in 1 mL chilled 0.1 M HCI containing 0.5% Triton X-100; incubate the suspension on ice for 10 minutes. Dilute acid with 5 mL distilled water and centrifuge at 200 x g for 10 min. Resuspend cells in 2 mL distilled water.
3. Denature cellular DNA by submerging the cell suspension into a boiling water bath for 10 min. Afterwards, quickly cool by placing the cell suspension in an ice slurry for several minutes. Wash cells in PBS that contains 0.5% Triton X-100.
4. Resuspend the cells (1-2 x 10 6 cells) in 100 μL of solution containing approximately 2 μg/mL anti-bromodeoxyuridine antibody diluted in PBS containing 0.1% BSA (0.2 μg/test). Incubate for 30 min at room temperature. Wash cells with PBS.
5. Resuspend cells in 100 μL of diluted goat anti-mouse IgG-FlTC Wash cells with PBS.
APPLICATIONS (Cont.)
Immunohistochemistry: Below is a procedure for staining cells that have been labeled with BrdU in vivo or in vitro. The procedure is based on the methods of B. Schutte et al. (1987) and D. Campana et al. (1988).
Preparation of tissue:
Inject animal with 50 mg BrdU/kg body weight. Sacrifice animal one hour later and remove organ or tissue under study. Embed tissue in OCT medium and snap-freeze by immersion into liquid nitrogen.Cut 4 mm frozen sections with a cryostat. Place sections on either albumin- or gelatin-coated slides.
Preparation of cells:
Pulse cells with 10 mM BrdU for 60 min. Cells grown on coverslips, or cytocentrifuge preparations made from cells grown in suspension, can be used for anti-bromodeoxyuridine staining according to the procedure below.
Procedure
1. Fix tissue sections or cells (on slide or coverglass) by immersing in absolute methanol for 10 minutes at +2-8°C. Air dry after removing from fixative. The slides can be stored at -20°C in a sealed box, or rehydrated to prepare for the assay procedure. To rehydrate, immerse in PBS for 3 min.
2. Denature DNA by incubating the slides in 2 N HCI for 60 min at +37°C.
3. Neutralize the acid by immersing the slides in 0.1 M borate buffer, pH 8.5. Change the buffer twice over a 10 min period.
4. Wash slides with PBS, changing the solution three times over a 10 min period.
5. Place slides in a humidified chamber (e.g., a sealed plastic box layered with wet paper towels) and cover cells with 150-300 μL of solution containing approximately 6 μg/mL anti-bromodeoxyuridine antibody diluted in PBS with 0.1% BSA. Incubate for 60 min at room temperature.
6. Wash slides with PBS, changing the solution three times over a 10 min period.
7. Apply optimal dilution of a second antibody conjugate (e.g., anti-mouse IgG-peroxidase), incubate, wash, and perform detection with a substrate that produces an insoluble product. After detection, counterstain with Harris-modified hematoxylin if desired. Slides can then be dehydrated and mounted.
抗BrdU抗体 クローンAH4H7-1／131-14871（マウスモノクローナル抗体）は、FC、ICC、IHCにおいて検証済みであり、ブロモデオキシウリジン（別名:BrdU）を検出できます。
研究のカテゴリ
エピジェネティクスおよび核機能
エピジェネティクスおよび核機能
研究のサブカテゴリ
細胞周期、DNA複製および修復
細胞周期、DNA複製および修復
ターゲットの説明
検出されるブロモデオキシウリジン取込みタンパク質の分子量に依存します。
関連事項
先発品：MAB1467
物理的形状
150 mM塩化ナトリウムと0.1%アジ化ナトリウムを含有する10 mMリン酸緩衝液（pH 7.4）に溶解
フォーマット：精製品
精製プロテインA
保管および安定性
保存期間は-20℃で出荷日から6ヵ月間です。凍結・融解サイクルの繰り返しを避けるため、分注してください。製品を最大限回収するために、溶解後のキャップを外す前に元のバイアルを遠心分離します。
アナリシスノート
対照
BrdU取り込み後のすべてのDNA含有分子種
BrdU取り込み後のすべてのDNA含有分子種
その他情報
濃度：ロットの具体的な濃度につきましては分析証明書をご参照ください。
法的情報
CHEMICON is a registered trademark of Merck KGaA, Darmstadt, Germany
免責事項
メルクのカタログまたは製品に添付されたメルクのその他の文書に記載されていない場合、メルクの製品は研究用途のみを目的としているため、他のいかなる目的にも使用することはできません。このような目的としては、未承認の商業用途、in vitroの診断用途、ex vivoあるいはin vivoの治療用途、またはヒトあるいは動物へのあらゆる種類の消費あるいは適用などがありますが、これらに限定されません。
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保管分類コード
12 - Non Combustible Liquids
WGK
WGK 2
引火点(°F)
Not applicable
引火点(℃)
Not applicable
適用法令
試験研究用途を考慮した関連法令を主に挙げております。化学物質以外については、一部の情報のみ提供しています。 製品を安全かつ合法的に使用することは、使用者の義務です。最新情報により修正される場合があります。WEBの反映には時間を要することがあるため、適宜SDSをご参照ください。
Jan Code
MAB3424:
試験成績書（COA）
製品のロット番号・バッチ番号を入力して、試験成績書（COA） を検索できます。ロット番号・バッチ番号は、製品ラベルに「Lot」または「Batch」に続いて記載されています。
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