Anti-Glypican 3 抗体 [SP86] (ab95363)
Key features and details
- Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
- Rabbit monoclonal [SP86] to Glypican 3
- Suitable for: ICC/IF, Flow Cyt, WB, IHC-P
- Knockout validated
- Reacts with: Human
Related conjugates and formulations
製品の概要
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製品名
Anti-Glypican 3 antibody [SP86]
Glypican 3 一次抗体 製品一覧 -
製品の詳細
Rabbit monoclonal [SP86] to Glypican 3 -
由来種
Rabbit -
アプリケーション
適用あり: ICC/IF, Flow Cyt, WB, IHC-Pmore details -
種交差性
交差種: Human -
免疫原
Synthetic peptide within Human Glypican 3 aa 500 to the C-terminus (C terminal). The exact sequence is proprietary.
Database link: P51654 -
ポジティブ・コントロール
- Human liver cancer tissueThis antibody gave a positive result when used in the following formaldehyde fixed cell lines: HepG2. ICC/IF: HepG2 Flow Cyt: HepG2, Hap1 cells
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特記事項
This product is a recombinant monoclonal antibody, which offers several advantages including:
- - High batch-to-batch consistency and reproducibility
- - Improved sensitivity and specificity
- - Long-term security of supply
- - Animal-free production
Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMAb® patents.
This product is FOR RESEARCH USE ONLY. For commercial use, please contact partnerships@abcam.com.
製品の特性
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製品の状態
Liquid -
保存方法
Shipped at 4°C. Upon delivery aliquot and store at -20°C. Avoid freeze / thaw cycles. -
バッファー
pH: 7.20
Preservative: 0.1% Sodium azide
Constituents: 1% BSA, PBS -
Concentration information loading...
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精製度
Protein A purified -
ポリ/モノ
モノクローナル -
クローン名
SP86 -
アイソタイプ
IgG -
研究分野
関連製品
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Alternative Versions
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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ab95363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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ICC/IF |
1/200.
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Flow Cyt |
1/80.
ab172730 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody. |
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WB |
Use at an assay dependent concentration. Predicted molecular weight: 66 kDa.
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IHC-P |
1/100. Perform heat mediated antigen retrieval (boil tissue section in 10mM citrate buffer, pH 6.0 for 10 min followed by cooling at RT for 20 min).
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特記事項 |
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ICC/IF
1/200. |
Flow Cyt
1/80. ab172730 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody. |
WB
Use at an assay dependent concentration. Predicted molecular weight: 66 kDa. |
IHC-P
1/100. Perform heat mediated antigen retrieval (boil tissue section in 10mM citrate buffer, pH 6.0 for 10 min followed by cooling at RT for 20 min). |
ターゲット情報
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機能
Cell surface proteoglycan that bears heparan sulfate. Inhibits the dipeptidyl peptidase activity of DPP4. May be involved in the suppression/modulation of growth in the predominantly mesodermal tissues and organs. May play a role in the modulation of IGF2 interactions with its receptor and thereby modulate its function. May regulate growth and tumor predisposition. -
組織特異性
Highly expressed in lung, liver and kidney. -
関連疾患
Defects in GPC3 are the cause of Simpson-Golabi-Behmel syndrome type 1 (SGBS1) [MIM:312870]; also known as Simpson dysmorphia syndrome (SDYS). SGBS is a condition characterized by pre- and postnatal overgrowth (gigantism) with visceral and skeletal anomalies. -
配列類似性
Belongs to the glypican family. -
細胞内局在
Cell membrane and Secreted > extracellular space. - Information by UniProt
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参照データベース
- Entrez Gene: 2719 Human
- Omim: 300037 Human
- SwissProt: P51654 Human
- Unigene: 644108 Human
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別名
- DGSX antibody
- Glypican proteoglycan 3 antibody
- Glypican-3 [Precursor] antibody
see all
画像
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Flow cytometry overlay histogram showing left wild-type Hap1 positive cells and right negative GPC3 knockout Hap1 stained with ab95363 (red line). The cells were incubated in 1x PBS containing 10% normal goat serum to block non-specific protein-protein interaction followed by the antibody (ab95363) (1x 106 in 100μl at 5.0 μg/ml (1/396)) for 30min on ice.
The secondary antibody Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed was incubated at 1/4000 for 30min on ice
Isotype control antibody Recombinant Rabbit IgG, monoclonal [EPR25A] - Isotype Control (black line) was used at the same concentration and conditions as the primary antibody. Unlabelled sample was also used as a control (blue line).
Acquisition of >5000 events were collected using a 50 mW Blue laser (488nm) and 525/40 bandpass filter.
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Lane 1: Wild-type HAP1 whole cell lysate (20 µg)
Lane 2: GPC3 knockout HAP1 whole cell lysate (20 µg)
Lane 3: HepG2 whole cell lysate (20 µg)Lanes 1 - 3: Merged signal (red and green). Green - ab95363 observed at 70 kDa. Red - loading control, ab130007, observed at 125kDa.
ab95363 was shown to specifically react with Glypican 3 in wild-type HAP1 cells as signal was lost in GPC3 knockout cells. Wild-type and GPC3 knockout samples were subjected to SDS-PAGE. ab95363 and ab130007 (Mouse anti-vinculin loading control) were incubated overnight at 4°C at 1/1000 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 1/10000 dilution for 1 hour at room temperature before imaging. -
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of Human hepatocellular carcinoma tissue sections labeling Glypican 3 with ab95363 at 1:100 dilution (4.66 ?g/ml). Heat mediated antigen retrieval with sodium citrate buffer (pH 6.0, epitope retrieval solution 1) for 10mins. Rabbit specific IHC polymer detection kit HRP/DAB (ab209101) was used as the secondary antibody. Hematoxylin was used as a counterstain. Positive staining on human hepatocellular carcinoma, performed on a Leica Biosystems BONDTM RX instrument.
The section was incubated with ab95363 for 30 mins at room temperature. -
Immunocytochemistry/ Immunofluorescence analysis of HepG2 (human hepatocellular carcinoma epithelial cell) cells labeling Glypican 3 with purified ab95363 at 1/200 (2.3 µg/ml). Cells were fixed in 100% Methanol and permeabilized with None. Cells were counterstained with Ab195889 Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor® 594) 1/200 (2.5 µg/ml). Goat anti rabbit IgG (Alexa Fluor® 488, ab150077) was used as the secondary antibody at 1/1000 (2 µg/ml) dilution. DAPI was used as nuclear counterstain. PBS instead of the primary antibody was used as the secondary antibody only control.
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Flow cytometry analysis of HepG2 (human hepatocellular carcinoma) labeling Glypican 3 with purified ab95363 at 1/80 dilution (5.825 µg/ml) (red). Goat anti rabbit IgG (Alexa Fluor® 488, ab150077) at 1/2000 dilution was used as a secondary antibody. Isotype control - Rabbit monoclonal IgG (ab172730) (Black). Unlableled control -Unlabelled cells (blue).
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Immunohistochemical staining of human liver hepatocellular carcinoma with ab95363.
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Flow cytometric analysis of rabbit anti-Glypican 3 (SP86) antibody ab98363 (1/100) in HEPG2 cellls (green) compared to negative control of rabbit IgG (blue).
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ab95363, at 1/100 dilution, staining Glypican 3 in formalin-fixed, paraffin-embedded Human liver cancer tissue by Immunohistochemistry.
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ICC/IF image of ab95363 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 ab95363 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.
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 (6)
ab95363 は 6 報の論文で使用されています。
- Jiao J et al. Galectin-9 expression predicts poor prognosis in hepatitis B virus-associated hepatocellular carcinoma. Aging (Albany NY) 14:1879-1890 (2022). PubMed: 35202002
- Jiang Z et al. Transforming primary human hepatocytes into hepatocellular carcinoma with genetically defined factors. EMBO Rep 23:e54275 (2022). PubMed: 35437924
- Sahu B et al. Human cell transformation by combined lineage conversion and oncogene expression. Oncogene 40:5533-5547 (2021). PubMed: 34302118
- Feng S et al. Multi-Modal Imaging Probe for Glypican-3 Overexpressed in Orthotopic Hepatocellular Carcinoma. J Med Chem 64:15639-15650 (2021). PubMed: 34590489
- Dong W et al. Prognostic Nomogram for Sorafenib Benefit in Hepatitis B Virus-Related Hepatocellular Carcinoma After Partial Hepatectomy. Front Oncol 10:605057 (2020). PubMed: 33643907
- Wang Y et al. Transcobalamin I: a novel prognostic biomarker of neoadjuvant chemotherapy in locally advanced hypopharyngeal squamous cell cancers. Onco Targets Ther 11:4253-4261 (2018). PubMed: 30100732