重组Anti-Glypican 3抗体[SP86] - BSA and Azide free (ab238804)
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 - BSA and Azide free
- Suitable for: IHC-P, ICC/IF, WB, Flow Cyt
- Knockout validated
- Reacts with: Human
Related conjugates and formulations
概述
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产品名称
Anti-Glypican 3抗体[SP86] - BSA and Azide free
参阅全部 Glypican 3 一抗 -
描述
兔单克隆抗体[SP86] to Glypican 3 - BSA and Azide free -
宿主
Rabbit -
经测试应用
适用于: IHC-P, ICC/IF, WB, Flow Cytmore details -
种属反应性
与反应: Human -
免疫原
Synthetic peptide. This information is proprietary to Abcam and/or its suppliers.
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阳性对照
- WB: HepG2 and Wild-type HAP1 whole cell lysate. ICC/IF: HepG2 cells Flow Cyt: HepG2, Hap1 cells
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常规说明
ab238804 is the carrier-free version of ab95363.
Our carrier-free antibodies are typically supplied in a PBS-only formulation, purified and free of BSA, sodium azide and glycerol. The carrier-free buffer and high concentration allow for increased conjugation efficiency.
This conjugation-ready format is designed for use with fluorochromes, metal isotopes, oligonucleotides, and enzymes, which makes them ideal for antibody labelling, functional and cell-based assays, flow-based assays (e.g. mass cytometry) and Multiplex Imaging applications.
Use our conjugation kits for antibody conjugates that are ready-to-use in as little as 20 minutes with <1 minute hands-on-time and 100% antibody recovery: available for fluorescent dyes, HRP, biotin and gold.
This product is compatible with the Maxpar® Antibody Labeling Kit from Fluidigm, without the need for antibody preparation. Maxpar® is a trademark of Fluidigm Canada Inc.
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
This product is FOR RESEARCH USE ONLY. For commercial use, please contact partnerships@abcam.com.
性能
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形式
Liquid -
存放说明
Shipped at 4°C. Store at +4°C. Do Not Freeze. -
存储溶液
pH: 7.20
Constituent: PBS -
无载体
是 -
Concentration information loading...
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纯度
Protein A/G purified -
纯化说明
Purified from TCS by protein A/G. -
克隆
单克隆 -
克隆编号
SP86 -
同种型
IgG -
研究领域
相关产品
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Alternative Versions
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Compatible Secondaries
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Conjugation kits
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Isotype control
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Related Products
应用
The Abpromise guarantee
Abpromise™承诺保证使用ab238804于以下的经测试应用
“应用说明”部分 下显示的仅为推荐的起始稀释度;实际最佳的稀释度/浓度应由使用者检定。
应用 | Ab评论 | 说明 |
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IHC-P |
Use at an assay dependent concentration.
Perform heat mediated antigen retrieval (Boil tissue section in 10 mM citrate buffer, pH 6.0 for 10 minutes followed by cooling at RT for 20 minutes). |
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ICC/IF |
Use at an assay dependent concentration.
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WB |
Use at an assay dependent concentration. Predicted molecular weight: 66 kDa.
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Flow Cyt |
Use at an assay dependent concentration.
ab172730 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody. |
说明 |
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IHC-P
Use at an assay dependent concentration. Perform heat mediated antigen retrieval (Boil tissue section in 10 mM citrate buffer, pH 6.0 for 10 minutes followed by cooling at RT for 20 minutes). |
ICC/IF
Use at an assay dependent concentration. |
WB
Use at an assay dependent concentration. Predicted molecular weight: 66 kDa. |
Flow Cyt
Use at an assay dependent concentration. ab172730 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody. |
靶标
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功能
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. -
组织特异性
Highly expressed in lung, liver and kidney. -
疾病相关
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. -
序列相似性
Belongs to the glypican family. -
细胞定位
Cell membrane and Secreted > extracellular space. - Information by UniProt
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数据库链接
- Entrez Gene: 2719 Human
- Omim: 300037 Human
- SwissProt: P51654 Human
- Unigene: 644108 Human
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别名
- DGSX antibody
- Glypican proteoglycan 3 antibody
- Glypican-3 [Precursor] antibody
see all
图片
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This image was generated using ab95363, the same clone, but with a different buffer formulation.
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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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 µg/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 BOND™ RX instrument.
The section was incubated with ab95363 for 30 mins at room temperature.
This image was generated using ab95363, the same clone, but with a different buffer formulation.
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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.
This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab95363).
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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. Isotypecontrol - Rabbit monoclonal IgG (ab172730) (black). Unlableled control - Unlabelled cells (blue).
This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (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).
This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab95363).
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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.
This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab95363).
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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.This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA and sodium azide (ab95363).
实验方案
To our knowledge, customised protocols are not required for this product. Please try the standard protocols listed below and let us know how you get on.
数据表及文件
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Datasheet download
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