製品の概要
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製品名Anti-PARL antibody
PARL 一次抗体 製品一覧 -
製品の詳細Rabbit polyclonal to PARL
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由来種Rabbit
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アプリケーション適用あり: WB, IHC-P, ICC/IFmore details
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種交差性交差種: Mouse, Rat, Human
交差が予測される動物種: Cow, Orangutan
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免疫原
Synthetic peptide conjugated to KLH derived from within residues 350 to the C-terminus of Human PARL.
(Peptide available as ab45230.)
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ポジティブ･コントロール
- ab45231 gave a positive result in the following whole cell lysates: Hela (data not shown), Jurkat, A431, HEK293 (data not shown), MCF7, U20S (data not shown). IHC-P: human colon adenocarcinoma FFPE tissue sections This antibody gave a positive result when used in the following formaldehyde fixed cell lines: HepG2.
製品の特性
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製品の状態Liquid
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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.
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バッファーpH: 7.40
Preservative: 0.02% Sodium azide
Constituents: PBS, 1% BSA
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... -
精製度Immunogen affinity purified
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ポリ/モノポリクローナル
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アイソタイプIgG
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研究分野
関連製品
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Compatible Secondaries
アプリケーション
Our Abpromise guarantee covers the use of ab45231 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
| アプリケーション | Abreviews | 特記事項 |
|---|---|---|
| WB | Use a concentration of 1 µg/ml. Detects a band of approximately 36 kDa (predicted molecular weight: 42 kDa). | |
| IHC-P | Use a concentration of 1 µg/ml. Perform heat mediated antigen retrieval before commencing with IHC staining protocol. | |
| ICC/IF | Use a concentration of 5 µg/ml. |
ターゲット情報
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機能Required for the control of apoptosis during postnatal growth. Essential for proteolytic processing of an antiapoptotic form of OPA1 which prevents the release of mitochondrial cytochrome c in response to intrinsic apoptoptic signals (By similarity). Promotes changes in mitochondria morphology regulated by phosphorylation of P-beta domain.
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配列類似性Belongs to the peptidase S54 family.
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翻訳後修飾P-beta is proteolytically processed (beta-cleavage) in a PARL-dependent manner. The cleavage is inhibited when residues Ser-65, Thr-69 and Ser-70 are all phosphorylated.
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細胞内局在Mitochondrion inner membrane and Nucleus. Translocated into the nucleus by an unknown mechanism.
- Information by UniProt
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参照データベース
- Entrez Gene: 514191 Cow
- Entrez Gene: 55486 Human
- Entrez Gene: 381038 Mouse
- Entrez Gene: 287979 Rat
- Omim: 607858 Human
- SwissProt: Q2KHV4 Cow
- SwissProt: Q9H300 Human
- SwissProt: Q5XJY4 Mouse
see all -
別名
- Mitochondrial intramembrane cleaving protease PARL antibody
- Mitochondrial intramembrane-cleaving protease PARL antibody
- P-beta antibody
see all
画像
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Immunocytochemistry/ Immunofluorescence - Anti-PARL antibody (ab45231)ICC/IF image of ab45231 stained HepG2 cells. The cells were 4% formaldehyde fixed (10 min) and then incubated in 1%BSA / 10% normal goat serum / 0.3M glycine in 0.1% PBS-Tween for 1h to permeabilise the cells and block non-specific protein-protein interactions. The cells were then incubated with the antibody ab45231 at 5µg/ml overnight at +4°C. The secondary antibody (green) was DyLight® 488 goat anti- rabbit (ab96899) IgG (H+L) used at a 1/250 dilution for 1h. Alexa Fluor® 594 WGA was used to label plasma membranes (red) at a 1/200 dilution for 1h. DAPI was used to stain the cell nuclei (blue) at a concentration of 1.43µM.
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Western blot - Anti-PARL antibody (ab45231)All lanes : Anti-PARL antibody (ab45231) at 1 µg/ml
Lane 1 : Jurkat (Human T cell lymphoblast-like cell line) Whole Cell Lysate
Lane 2 : A431 (Human epithelial carcinoma cell line) Whole Cell Lysate
Lane 3 : MCF7 (Human breast adenocarcinoma cell line) Whole Cell Lysate
Lysates/proteins at 10 µg per lane.
Secondary
All lanes : IRDye 680 Conjugated Goat Anti-Rabbit IgG (H+L) at 1/10000 dilution
Performed under reducing conditions.
Predicted band size: 42 kDa
Observed band size: 36 kDa (why is the actual band size different from the predicted?)
Additional bands at: 150 kDa, 51 kDa, 90 kDa. We are unsure as to the identity of these extra bands.PARL is a mitochondrial protein which is constitutively cleaved, leaving a protein of 36.5 kDa (Sík A et al, J Biol Chem, 2004 Apr 9;279(15):15323-9 (PMID 14732705)). ab45231 detects a band at 36 kDa (cleaved form). However, Abcam's data suggests that ab45231 does not detect the full PARL protein (42 kDa).
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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-PARL antibody (ab45231)IHC image of PARL staining in human colon adenocarcinoma 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 ab45231, 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.
For other IHC staining systems (automated and non-automated) customers should optimize variable parameters such as antigen retrieval conditions, primary antibody concentration and antibody incubation times.
プロトコール
参考文献
This product has been referenced in:
- Shi G & McQuibban GA The Mitochondrial Rhomboid Protease PARL Is Regulated by PDK2 to Integrate Mitochondrial Quality Control and Metabolism. Cell Rep 18:1458-1472 (2017). Read more (PubMed: 28178523) »
- Meissner C et al. Intramembrane protease PARL defines a negative regulator of PINK1- and PARK2/Parkin-dependent mitophagy. Autophagy 11:1484-98 (2015). Read more (PubMed: 26101826) »
レビューと Q&A
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