OP46
Anti-MDM2 (Ab-1) Mouse mAb (IF2)
liquid, clone IF2, Calbiochem®
別名:
Anti-Murine Double Minute Chromosome-2, Anti-Ubiquitin Protein Ligase, Anti-p53 Binding Protein, Anti-Ubiquitin Protein Ligase, Anti-p53 Binding Protein, Anti-Murine Double Minute Chromosome-2
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About This Item
UNSPSCコード:
12352203
NACRES:
NA.41
クローン:
IF2, monoclonal
application:
化学種の反応性:
human
citations:
82
おすすめの製品
由来生物
mouse
品質水準
抗体製品の状態
purified antibody
抗体製品タイプ
primary antibodies
クローン
IF2, monoclonal
フォーム
liquid
含みます
≤0.1% sodium azide as preservative (100 μg only)
化学種の反応性
human
以下との反応禁止
mouse
メーカー／製品名
Calbiochem®
保管条件
do not freeze
アイソタイプ
IgG2b
輸送温度
wet ice
保管温度
2-8°C
ターゲットの翻訳後修飾
unmodified
遺伝子情報
human ... MDM2(4193)
mouse ... Mdm2(17246)
詳細
Purified mouse monoclonal antibody (see application references). Recognizes the ~90 kDa (apparent MW) MDM2 protein. Also recognizes isoforms at ~57 and ~74/76 kDa.
Recognizes the ~90 kDa (apparent MW) MDM2 protein. Also recognizes isoforms of ~57 kDa and ~74/76 kDa by immunoblotting.
This Anti-MDM2 (Ab-1) Mouse mAb (IF2) is validated for use in Frozen Sections Immunoblotting Immunofluorescence Immunoprecipitation Paraffin Sections for the detection of MDM2 (Ab-1).
免疫原
Epitope: within amino acids 26-169 of human MDM2
Human
human MDM2
アプリケーション
Frozen Sections (1-5 µg/ml, see application references)
Immunoblotting (0.5-2 µg/ml, chemiluminescence)
Immunofluorescence (1-5 µg/ml)
Immunoprecipitation (1 µg/sample)
Paraffin Sections (1-5 µg/ml, heat pre-treatment required, see application references)
Immunoblotting (0.5-2 µg/ml, chemiluminescence)
Immunofluorescence (1-5 µg/ml)
Immunoprecipitation (1 µg/sample)
Paraffin Sections (1-5 µg/ml, heat pre-treatment required, see application references)
包装
Please refer to vial label for lot-specific concentration.
警告
Toxicity: Standard Handling (A)
物理的形状
In 50 mM sodium phosphate buffer, 0.2% gelatin.
アナリシスノート
Positive Control
OSA-CL cells
OSA-CL cells
その他情報
Although the amino acid sequence of MDM2 predicts a protein with a molecular mass of approximately 54 kDa, MDM2 protein migrates on SDS/PAGE with an apparent mobility of 90 kDa.
Immunoblotting Protocol
MDM2 (Ab-1) can be used to detect MDM2 by Western blot of proteins previously separated by SDS/PAGE and electrophoretically transferred onto nitrocellulose membranes. The proteins are reacted with the monoclonal antibody and visualized using an HRP conjugated goat anti-mouse antibody with chemiluminescent detection.
Materials
Equipment:
• Electrophoresis apparatus
• Electroblotting apparatus
• Rocker platform
Solutions and Reagents
• Anti-MDM2 (Ab-1) Mouse mAb (IF2) Cat. No. OP46 or OP46T
• HRP conjugated goat anti-mouse IgG heavy and light chains (e.g. Cat. No. 401215)
• Chemiluminescence detection system
• ELB Buffer (include a cocktail of proetease inhibitors, such as 0.5 µg/ml leupeptin, 1 µg/ml pepstatin, 1 mM EDTA and 0.2 mM PMSF): 50 mM Hepes pH 7.0, 250 mM NaCl, 0.5 mM EDTA, 0.1% Nonidet P-40 Alternative
• SDS-PAGE (7% acrylamide)
• Phosphate buffered saline (PBS) pH 7.4; 1 Liter: 0.2 g KCl, 0.2 g KH2PO4, 8 g NaCl, 1.15 g Na2HPO4
• PBS/0.1% Tween®-20 detergent (PBST)
• 3% Non-fat Dry Milk in PBST
Procedure
1. Lyse cells in ELB Buffer. (Alternatively, cells can be lysed in RIPA Buffer or directly into 1x Laemmli Sample Buffer).
2. Electrophorese 50-100 µg lysate using a 7% acrylamide gel.
3. Transfer the protein samples from the polyacrylamide gel onto a nitrocellulose membrane using an electroblotting apparatus.
4. Block the membrane for 1 h in PBST containing 3% non-fat dry milk at room temperature with rocking. Use about 1 ml per cm2 of membrane.
5. Incubate the membrane with 1 µg/ml Anti-MDM2 (Ab-1) Mouse mAb (IF2) in 3% non-fat dry milk/ PBST for 1 h at room temperature with rocking.
6. Wash the membrane 3 times, 15 min each, in PBST at room temperature with rocking.
7. Incubate the membrane with HRP conjugated goat anti-mouse IgG heavy and light chain antibody, diluted according to the supplier’s instructions, in 3% non-fat dry milk/ PBST at room temperature for 1 h.
8. Wash the membrane 4 times, 15 min each, in PBST at room temperature with rocking.
9. Develop the membrane using chemiluminescent detection reagents according to manufacturer instructions.
10. Expose the membrane to film for ten minutes. Adjust subsequent exposure times as needed.
Immunoblotting Protocol
MDM2 (Ab-1) can be used to detect MDM2 by Western blot of proteins previously separated by SDS/PAGE and electrophoretically transferred onto nitrocellulose membranes. The proteins are reacted with the monoclonal antibody and visualized using an HRP conjugated goat anti-mouse antibody with chemiluminescent detection.
Materials
Equipment:
• Electrophoresis apparatus
• Electroblotting apparatus
• Rocker platform
Solutions and Reagents
• Anti-MDM2 (Ab-1) Mouse mAb (IF2) Cat. No. OP46 or OP46T
• HRP conjugated goat anti-mouse IgG heavy and light chains (e.g. Cat. No. 401215)
• Chemiluminescence detection system
• ELB Buffer (include a cocktail of proetease inhibitors, such as 0.5 µg/ml leupeptin, 1 µg/ml pepstatin, 1 mM EDTA and 0.2 mM PMSF): 50 mM Hepes pH 7.0, 250 mM NaCl, 0.5 mM EDTA, 0.1% Nonidet P-40 Alternative
• SDS-PAGE (7% acrylamide)
• Phosphate buffered saline (PBS) pH 7.4; 1 Liter: 0.2 g KCl, 0.2 g KH2PO4, 8 g NaCl, 1.15 g Na2HPO4
• PBS/0.1% Tween®-20 detergent (PBST)
• 3% Non-fat Dry Milk in PBST
Procedure
1. Lyse cells in ELB Buffer. (Alternatively, cells can be lysed in RIPA Buffer or directly into 1x Laemmli Sample Buffer).
2. Electrophorese 50-100 µg lysate using a 7% acrylamide gel.
3. Transfer the protein samples from the polyacrylamide gel onto a nitrocellulose membrane using an electroblotting apparatus.
4. Block the membrane for 1 h in PBST containing 3% non-fat dry milk at room temperature with rocking. Use about 1 ml per cm2 of membrane.
5. Incubate the membrane with 1 µg/ml Anti-MDM2 (Ab-1) Mouse mAb (IF2) in 3% non-fat dry milk/ PBST for 1 h at room temperature with rocking.
6. Wash the membrane 3 times, 15 min each, in PBST at room temperature with rocking.
7. Incubate the membrane with HRP conjugated goat anti-mouse IgG heavy and light chain antibody, diluted according to the supplier’s instructions, in 3% non-fat dry milk/ PBST at room temperature for 1 h.
8. Wash the membrane 4 times, 15 min each, in PBST at room temperature with rocking.
9. Develop the membrane using chemiluminescent detection reagents according to manufacturer instructions.
10. Expose the membrane to film for ten minutes. Adjust subsequent exposure times as needed.
Gorgoulis, V.G., et al. 1996. J. Pathol.180, 129.
Marchetti, A., et al. 1995. J. Pathol.175, 31.
Barak, Y., et al. 1993. EMBO J.12, 461.
Ladanyi, M., et al. 1993. Cancer Res.53, 16.
Leach, F.S., et al. 1993. Cancer Res.53, 2231.
Oliner, J.D., et al. 1993. Nature362, 857.
Momand, J., et al. 1992. Cell69, 1237.
Oliner, J.D., et al. 1992. Nature358, 80.
Fakharzadeh, S.S., et al. 1991. EMBO J. 10, 1565.
Marchetti, A., et al. 1995. J. Pathol.175, 31.
Barak, Y., et al. 1993. EMBO J.12, 461.
Ladanyi, M., et al. 1993. Cancer Res.53, 16.
Leach, F.S., et al. 1993. Cancer Res.53, 2231.
Oliner, J.D., et al. 1993. Nature362, 857.
Momand, J., et al. 1992. Cell69, 1237.
Oliner, J.D., et al. 1992. Nature358, 80.
Fakharzadeh, S.S., et al. 1991. EMBO J. 10, 1565.
法的情報
CALBIOCHEM is a registered trademark of Merck KGaA, Darmstadt, Germany
TWEEN is a registered trademark of Croda International PLC
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保管分類コード
10 - Combustible liquids
WGK
nwg
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
OP46-100UG:
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。
Yongliang Hu et al.
Oncogene, 38(5), 731-746 (2018-09-05)
Our previous studies revealed that GADD45α is a liable protein, which undergoes MDM2-dependent constitutive ubiquitination and degradation in resting HepG2 hepatoma cells. Arsenite exposure induces ribosomal stress responses mediated by the ribosomal protein S7, which can block MDM2 activity and
C Wasylyk et al.
Molecular and cellular biology, 20(15), 5554-5570 (2000-07-13)
The cell cycle arrest and proapoptotic functions of p53 are under tight control by Mdm2. After stress activation of p53 by nontranscriptional mechanisms, transcription of the mdm2 gene results in increased synthesis of Mdm2 and down-regulation of p53. Disruption of
Sharon Brookes et al.
The EMBO journal, 21(12), 2936-2945 (2002-06-18)
The CDKN2A tumour suppressor locus encodes two distinct proteins, p16(INK4a) and p14(ARF), both of which have been implicated in replicative senescence, the state of permanent growth arrest provoked in somatic cells by aberrant proliferative signals or by cumulative population doublings
Deborah J Luessen et al.
The Journal of biological chemistry, 294(38), 14068-14080 (2019-08-02)
Acute alcohol exposure alters the trafficking and function of many G-protein-coupled receptors (GPCRs) that are associated with aberrant behavioral responses to alcohol. However, the molecular mechanisms underlying alcohol-induced changes in GPCR function remain unclear. β-Arrestin is a key player involved
Jennifer Hüllein et al.
Cancer research, 79(12), 3125-3138 (2019-04-20)
Oncogenic MYC activation promotes proliferation in Burkitt lymphoma, but also induces cell-cycle arrest and apoptosis mediated by p53, a tumor suppressor that is mutated in 40% of Burkitt lymphoma cases. To identify molecular dependencies in Burkitt lymphoma, we performed RNAi-based
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