Anti-ACSS2 抗体 (ab66038)
Key features and details
- Rabbit polyclonal to ACSS2
- Suitable for: WB, IHC-P, IP
- Knockout validated
- Reacts with: Human
- Isotype: IgG
リコンビナント抗体で、ロット間での高い再現性を実現
- 異なるロット間での安定した再現性
- 容易なスケールアップ
- 評価試験による特異性の確認済み
- 倫理基準に準拠 - アニマル・フリーの生産
製品の概要
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製品名
Anti-ACSS2 antibody
ACSS2 一次抗体 製品一覧 -
製品の詳細
Rabbit polyclonal to ACSS2 -
由来種
Rabbit -
特異性
Replenishment batches of our polyclonal antibody, ab66038 are tested in WB. Previous batches were additionally validated in IHC-P and IP. These applications are still expected to work and are covered by our Abpromise guarantee. You may also be interested in our alternative recombinant antibody, ab133664.
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アプリケーション
適用あり: WB, IHC-P, IPmore details
適用なし: ICC/IF -
種交差性
交差種: Human
交差が予測される動物種: Mouse, Rat -
免疫原
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ポジティブ・コントロール
- This antibody gave a positive signal in Human Liver and Human Colon Tissue lysates and in the following whole cell lysates: HepG2; U-87 MG; Caco 2.
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特記事項
The Life Science industry has been in the grips of a reproducibility crisis for a number of years. Abcam is leading the way in addressing this with our range of recombinant monoclonal antibodies and knockout edited cell lines for gold-standard validation. Please check that this product meets your needs before purchasing.
If you have any questions, special requirements or concerns, please send us an inquiry and/or contact our Support team ahead of purchase. Recommended alternatives for this product can be found below, along with publications, customer reviews and Q&As
製品の特性
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製品の状態
Liquid -
保存方法
Shipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C or -80°C. Avoid freeze / thaw cycle. -
バッファー
pH: 7.40
Preservative: 0.02% Sodium azide
Constituent: PBS
Batches of this product that have a concentration < 1mg/ml may have BSA added as a stabilising agent. If you would like information about the formulation of a specific lot, please contact our scientific support team who will be happy to help. -
Concentration information loading...
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精製度
Immunogen affinity purified -
ポリ/モノ
ポリクローナル -
アイソタイプ
IgG -
研究分野
関連製品
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Compatible Secondaries
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Isotype control
アプリケーション
The Abpromise guarantee
Abpromise保証は、 次のテスト済みアプリケーションにおけるab66038の使用に適用されます
アプリケーションノートには、推奨の開始希釈率がありますが、適切な希釈率につきましてはご検討ください。
アプリケーション | Abreviews | 特記事項 |
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WB | (1) |
Use a concentration of 1 µg/ml. Detects a band of approximately 79 kDa (predicted molecular weight: 79 kDa).
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IHC-P |
Use a concentration of 5 µg/ml. Perform heat mediated antigen retrieval before commencing with IHC staining protocol.
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IP | (1) |
Use at an assay dependent concentration.
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特記事項 |
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WB
Use a concentration of 1 µg/ml. Detects a band of approximately 79 kDa (predicted molecular weight: 79 kDa). |
IHC-P
Use a concentration of 5 µg/ml. Perform heat mediated antigen retrieval before commencing with IHC staining protocol. |
IP
Use at an assay dependent concentration. |
ターゲット情報
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機能
Activates acetate so that it can be used for lipid synthesis or for energy generation. -
配列類似性
Belongs to the ATP-dependent AMP-binding enzyme family. -
細胞内局在
Cytoplasm. - Information by UniProt
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参照データベース
- Entrez Gene: 55902 Human
- Entrez Gene: 60525 Mouse
- Entrez Gene: 311569 Rat
- Omim: 605832 Human
- SwissProt: Q9NR19 Human
- SwissProt: Q8BK97 Mouse
- SwissProt: Q9QXG4 Mouse
- SwissProt: D3ZFF9 Rat
see all -
別名
- ACAS2 antibody
- AceCS antibody
- Acetate CoA ligase antibody
see all
画像
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All lanes : Anti-ACSS2 antibody (ab66038) at 1 µg/ml
Lane 1 : Wild-type HAP1 whole cell lysate
Lane 2 : ACSS2 knockout HAP1 whole cell lysate
Lane 3 : HepG2 whole cell lysate
Lane 4 : U-87 MG whole cell lysate
Lysates/proteins at 20 µg per lane.
Predicted band size: 79 kDa
Observed band size: 80 kDa why is the actual band size different from the predicted?Lanes 1 - 4: Merged signal (red and green). Green - ab66038 observed at 80 kDa. Red - loading control, ab8245, observed at 37 kDa.
ab66038 was shown to recognize ACSS2 in wild-type HAP1 cells as signal was lost at the expected MW in ACSS2 knockout cells. Additional cross-reactive bands were observed in the wild-type and knockout cells. Wild-type and ACSS2 knockout samples were subjected to SDS-PAGE. The membrane was blocked with 3% Milk. Ab66038 and ab8245 (Mouse anti GAPDH loading control) were incubated overnight at 4°C at 1 ug/ml and 1/20000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye® 800CW) preabsorbed ab216773 and Goat anti-Mouse IgG H&L (IRDye® 680RD) preabsorbed ab216776 secondary antibodies at 1/20000 dilution for 1 hour at room temperature before imaging.
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All lanes : Anti-ACSS2 antibody (ab66038) at 1 µg/ml
Lane 1 : Human liver tissue lysate - total protein (ab29889)
Lane 2 : HepG2 (Human hepatocellular liver carcinoma cell line) Whole Cell Lysate
Lane 3 : U-87 MG (Human glioblastoma astrocytoma) Whole Cell Lysate
Lane 4 : Human colon tissue lysate - total protein (ab30051)
Lane 5 : Caco-2 whole cell lysate (ab3950)
Lysates/proteins at 10 µg per lane.
Secondary
All lanes : Goat polyclonal to Rabbit IgG - H&L - Pre-Adsorbed (HRP) at 1/3000 dilution
Performed under reducing conditions.
Predicted band size: 79 kDa
Observed band size: 79 kDa
Additional bands at: 100 kDa, 38 kDa, 45 kDa. We are unsure as to the identity of these extra bands.
Exposure time: 30 seconds -
ACSS2 was immunoprecipitated using 0.5mg HepG2 whole cell extract, 5µg of Rabbit polyclonal to ACSS2 and 50µl of protein G magnetic beads (+). No antibody was added to the control (-).
The antibody was incubated under agitation with Protein G beads for 10min, HepG2 whole cell extract lysate diluted in RIPA buffer was added to each sample and incubated for a further 10min under agitation.
Proteins were eluted by addition of 40µl SDS loading buffer and incubated for 10min at 70oC; 10µl of each sample was separated on a SDS PAGE gel, transferred to a nitrocellulose membrane, blocked with 5% BSA and probed with ab66038.
Secondary: Mouse monoclonal [SB62a] Secondary Antibody to Rabbit IgG light chain (HRP) (ab99697).
Band: 79kDa: ACSS2. -
IHC image of ACSS2 staining in human liver carcinoma formalin fixed paraffin embedded tissue section, performed on a Leica BondTM system using the standard protocol F. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH6, epitope retrieval solution 1) for 20 mins. The section was then incubated with ab66038, 5µg/ml, for 15 mins at room temperature and detected using an HRP conjugated compact polymer system. DAB was used as the chromogen. The section was then counterstained with haematoxylin and mounted with DPX.
プロトコール
データシートおよび資料
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SDS download
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Datasheet download
参考文献 (8)
ab66038 は 8 報の論文で使用されています。
- Yelek C et al. Acetate: Friend or foe against breast tumour growth in the context of obesity? J Cell Mol Med 24:14195-14204 (2020). PubMed: 33107196
- Wang T et al. SENP1-Sirt3 Signaling Controls Mitochondrial Protein Acetylation and Metabolism. Mol Cell 75:823-834.e5 (2019). PubMed: 31302001
- Kappler M et al. Causes and Consequences of A Glutamine Induced Normoxic HIF1 Activity for the Tumor Metabolism. Int J Mol Sci 20:N/A (2019). PubMed: 31554283
- Shah A et al. TGIF transcription factors repress acetyl CoA metabolic gene expression and promote intestinal tumor growth. Genes Dev 33:388-402 (2019). PubMed: 30808659
- Sun L et al. Decreased expression of acetyl-CoA synthase 2 promotes metastasis and predicts poor prognosis in hepatocellular carcinoma. Cancer Sci 108:1338-1346 (2017). WB, IHC . PubMed: 28387999
- Chen R et al. Coordinate regulation of stress signaling and epigenetic events by Acss2 and HIF-2 in cancer cells. PLoS One 12:e0190241 (2017). PubMed: 29281714
- Chen R et al. The acetate/ACSS2 switch regulates HIF-2 stress signaling in the tumor cell microenvironment. PLoS One 10:e0116515 (2015). WB . PubMed: 25689462
- Xu M et al. An acetate switch regulates stress erythropoiesis. Nat Med 20:1018-26 (2014). PubMed: 25108527