Heparan Sulfate Proteoglycan 2/Perlecan重组抗体[RM2059]_Heparan Sulfate Proteoglycan 2/Perlecan抗体(ab318285)

产品名称

Anti-Heparan Sulfate Proteoglycan 2/Perlecan抗体[RM2059] - BSA and Azide free
参阅全部 Heparan Sulfate Proteoglycan 2/Perlecan 一抗
描述

兔重组multiclonal [RM2059] to Heparan Sulfate Proteoglycan 2/Perlecan - BSA and Azide free
宿主

Rabbit
特异性

Unsuitable for rat WB.
经测试应用

适用于: IHC-P, ICC/IF, Flow Cyt (Intra), IP, WBmore details
种属反应性

与反应: Mouse, Rat, Human
免疫原

This product was produced with the following immunogens:
Recombinant fragment. This information is proprietary to Abcam and/or its suppliers.

Recombinant fragment. This information is proprietary to Abcam and/or its suppliers.
阳性对照
- WB: HUVEC, EA.hy926, MEF and NIH/3T3 lysates. IHC-P: Human kidney, Human colon, Human colorectal carcinoma, Mouse kidney and Rat kidney tissues. ICC/IF: HUVEC and MEF cells. Flow Cyt (Intra): HUVEC, Neuro-2a and MEF cells. IP: EA.hy926 cell.
常规说明
ab318285 is the carrier-free version of ab318284
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
For more information see here.
Our RabMAb^® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMAb^® patents.

形式

Liquid
存放说明

Shipped at 4°C. Store at +4°C.
存储溶液

pH: 7.20
Constituent: 100% PBS
无载体

是
Concentration information loading...
纯度

Protein A purified
克隆

Recombinant Multiclonal
克隆编号

RM2059
同种型

IgG
研究领域

Alternative Versions
- Anti-Heparan Sulfate Proteoglycan 2/Perlecan antibody [RM2059] (ab318284)
Compatible Secondaries
Isotype control
- Rabbit IgG, monoclonal [EPR25A] - Isotype Control (ab172730)
Related Products
- Anti-Vinculin antibody [EPR8185] (ab129002)
- Alexa Fluor® 594 Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (ab195889)

The Abpromise guarantee

Abpromise™承诺保证使用ab318285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
IHC-P		Use at an assay dependent concentration. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
ICC/IF		Use at an assay dependent concentration.
Flow Cyt (Intra)		Use at an assay dependent concentration.
IP		Use at an assay dependent concentration.
WB		Use at an assay dependent concentration.

说明
IHC-P Use at an assay dependent concentration. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
ICC/IF Use at an assay dependent concentration.
Flow Cyt (Intra) Use at an assay dependent concentration.
IP Use at an assay dependent concentration.
WB Use at an assay dependent concentration.

功能

Integral component of basement membranes. Component of the glomerular basement membrane (GBM), responsible for the fixed negative electrostatic membrane charge, and which provides a barrier which is both size- and charge-selective. It serves as an attachment substrate for cells. Plays essential roles in vascularization. Critical for normal heart development and for regulating the vascular response to injury. Also required for avascular cartilage development.
Endorepellin in an anti-angiogenic and anti-tumor peptide that inhibits endothelial cell migration, collagen-induced endothelial tube morphogenesis and blood vessel growth in the chorioallantoic membrane. Blocks endothelial cell adhesion to fibronectin and type I collagen. Anti-tumor agent in neovascularization. Interaction with its ligand, integrin alpha2/beta1, is required for the anti-angiogenic properties. Evokes a reduction in phosphorylation of receptor tyrosine kinases via alpha2/beta1 integrin-mediated activation of the tyrosine phosphatase, PTPN6.
The LG3 peptide has anti-angiogenic properties that require binding of calcium ions for full activity.
组织特异性

Found in the basement membranes.
疾病相关

Defects in HSPG2 are the cause of Schwartz-Jampel syndrome (SJS1) [MIM:255800]; a rare autosomal recessive disorder characterized by permanent myotonia (prolonged failure of muscle relaxation) and skeletal dysplasia, resulting in reduced stature, kyphoscoliosis, bowing of the diaphyses and irregular epiphyses.
Defects in HSPG2 are the cause of dyssegmental dysplasia Silverman-Handmaker type (DDSH) [MIM:224410]. The dyssegmental dysplasias are rare, autosomal recessive skeletal dysplasias with anisospondyly and micromelia. There are two recognized types: the severe, lethal DDSH and the milder Rolland-Desbuquois form. Individuals with DDSH also have a flat face, micrognathia, cleft palate and reduced joint mobility, and frequently have an encephalocoele. The endochondral growth plate is short, the calcospherites (which are spherical calcium-phosphorus crystals produced by hypertrophic chondrocytes) are unfused, and there is mucoid degeneration of the resting cartilage.
序列相似性

Contains 4 EGF-like domains.
Contains 22 Ig-like C2-type (immunoglobulin-like) domains.
Contains 11 laminin EGF-like domains.
Contains 3 laminin G-like domains.
Contains 3 laminin IV type A domains.
Contains 4 LDL-receptor class A domains.
Contains 1 SEA domain.
翻译后修饰

Proteolytic processing produces the C-terminal angiogenic peptide, endorepellin. This peptide can be further processed to produce the LG3 peptide.
N- and O-glycosylated; contains three heparan sulfate chains. The LG3 peptide contains at least three and up to five potential O-glycosylation sites but no N-glycosylation.
细胞定位

Secreted > extracellular space > extracellular matrix > basement membrane.
Target information above from: UniProt accession P98160 The UniProt Consortium
The Universal Protein Resource (UniProt) in 2010
Nucleic Acids Res. 38:D142-D148 (2010) .

Information by UniProt
数据库链接
- Entrez Gene: 3339 Human
- Entrez Gene: 15530 Mouse
- Entrez Gene: 313641 Rat
- Omim: 142461 Human
- SwissProt: P98160 Human
- SwissProt: Q05793 Mouse
- Unigene: 562227 Human
- Unigene: 273662 Mouse
别名
- Basement membrane specific heparan sulfate proteoglycan core protein antibody
- Endorepellin (domain V region) antibody
- Heparan sulfate proteoglycan of basement membrane antibody
- HSPG 2 antibody
- HSPG antibody
- Hspg2 antibody
- LG3 peptide antibody
- Perlecan antibody
- PGBM_HUMAN antibody
- PLC antibody
- Schwartz Jampel syndrome 1 (chondrodystrophic myotonia) antibody
- SJA antibody
- SJS antibody
- SJS1 antibody
see all

Western blot - Anti-Heparan Sulfate Proteoglycan 2/Perlecan antibody [RM2059] - BSA and Azide free (ab318285)

All lanes : Anti-Heparan Sulfate Proteoglycan 2/Perlecan antibody [RM2059] (ab318284) at 1/1000 dilution

Lane 1 : HUVEC (human umbilical vein endothelial cell) whole cell lysate
Lane 2 : SH-SY5Y (human neuroblastoma epithelial cell) whole cell lysate
Lane 3 : EA.hy926 (human somatic cell hybrid endothelial cell) whole cell lysate

Lysates/proteins at 20 µg per lane.

Secondary
All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/20000 dilution

Observed band size: 469 kDa why is the actual band size different from the predicted?

Exposure time: 37 seconds

This data was developed using ab318284, the same antibody clone in a different buffer formulation.

Blocking and diluting buffer and concentration: 5% NFDM/TBST.

Low expression: SH-SY5Y (PMID:25883214).

In Western blot, Anti-Vinculin antibody [EPR8185] (ab129002) staining at 1/10000 dilution.
Western blot - Anti-Heparan Sulfate Proteoglycan 2/Perlecan antibody [RM2059] - BSA and Azide free (ab318285)

All lanes : Anti-Heparan Sulfate Proteoglycan 2/Perlecan antibody [RM2059] (ab318284) at 1/1000 dilution

Lane 1 : MEF (mouse embryo fibroblast) whole cell lysate
Lane 2 : Neuro-2a (mouse neuroblastoma neuroblast) whole cell lysate
Lane 3 : NIH/3T3 (mouse embryonic fibroblast) whole cell lysate
Lane 4 : RAW 264.7 (mouse abelson murine leukemia virus-induced tumor macrophage) whole cell lysate
Lane 5 : J774A.1 (mouse reticulum cell sarcoma monocyte/macrophage ) whole cell lysate

Lysates/proteins at 20 µg per lane.

Secondary
All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/20000 dilution

Developed using the ECL technique.

Observed band size: 469 kDa why is the actual band size different from the predicted?

Exposure time: 169 seconds

This data was developed using ab318284, the same antibody clone in a different buffer formulation.

Blocking and diluting buffer and concentration: 5% NFDM/TBST.

Low expression: RAW 264.7, J774A.1(PMID: 35887248), Neuro-2a.

The bands beneath the target band (469 kDa) are likely to be degraded target fragments.

In Western blot, Anti-Vinculin antibody [EPR8185] (ab129002) staining at 1/10000 dilution.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Heparan Sulfate Proteoglycan 2/Perlecan antibody [RM2059] - BSA and Azide free (ab318285)

This data was developed using ab318284, the same antibody clone in a different buffer formulation.

Immunohistochemical analysis of paraffin-embedded Human kidney tissue labeling Heparan Sulfate Proteoglycan 2/Perlecan with ab318284 at 1/1000 (0.486 ug/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Positive staining on basement membrane of human kidney.
The section was incubated with ab318284 for 30 mins at room temperature.
The immunostaining was performed on a Leica Biosystems BOND™ RX instrument

Counterstained with Hematoxylin.

Secondary antibody only control: Secondary antibody is a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Heat mediated antigen retrieval was performed with Tris-EDTA buffer (pH 9.0, Epitope Retrieval Solution2) for 20 mins
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Heparan Sulfate Proteoglycan 2/Perlecan antibody [RM2059] - BSA and Azide free (ab318285)

This data was developed using ab318284, the same antibody clone in a different buffer formulation.

Immunohistochemical analysis of paraffin-embedded Human colon tissue labeling Heparan Sulfate Proteoglycan 2/Perlecan with ab318284 at 1/1000 (0.486 ug/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Positive staining on human colon.
The section was incubated with ab318284 for 30 mins at room temperature.
The immunostaining was performed on a Leica Biosystems BOND™ RX instrument

Counterstained with Hematoxylin.

Secondary antibody only control: Secondary antibody is a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Heat mediated antigen retrieval was performed with Tris-EDTA buffer (pH 9.0, Epitope Retrieval Solution2) for 20 mins
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Heparan Sulfate Proteoglycan 2/Perlecan antibody [RM2059] - BSA and Azide free (ab318285)

This data was developed using ab318284, the same antibody clone in a different buffer formulation.

Immunohistochemical analysis of paraffin-embedded Human colorectal carcinoma tissue labeling Heparan Sulfate Proteoglycan 2/Perlecan with ab318284 at 1/1000 (0.486 ug/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Positive staining on human colorectal carcinoma.
The section was incubated with ab318284 for 30 mins at room temperature.
The immunostaining was performed on a Leica Biosystems BOND™ RX instrument

Counterstained with Hematoxylin.

Secondary antibody only control: Secondary antibody is a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Heat mediated antigen retrieval was performed with Tris-EDTA buffer (pH 9.0, Epitope Retrieval Solution2) for 20 mins
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Heparan Sulfate Proteoglycan 2/Perlecan antibody [RM2059] - BSA and Azide free (ab318285)

This data was developed using ab318284, the same antibody clone in a different buffer formulation.

Immunohistochemical analysis of paraffin-embedded Mouse kidney tissue labeling Heparan Sulfate Proteoglycan 2/Perlecan with ab318284 at 1/1000 (0.486 ug/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Positive staining on basement membrane of mouse kidney (PMID: 23757342).
The section was incubated with ab318284 for 30 mins at room temperature.
The immunostaining was performed on a Leica Biosystems BOND™ RX instrument

Counterstained with Hematoxylin.

Secondary antibody only control: Secondary antibody is a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Heat mediated antigen retrieval was performed with Tris-EDTA buffer (pH 9.0, Epitope Retrieval Solution2) for 20 mins
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Heparan Sulfate Proteoglycan 2/Perlecan antibody [RM2059] - BSA and Azide free (ab318285)

This data was developed using ab318284, the same antibody clone in a different buffer formulation.

Immunohistochemical analysis of paraffin-embedded Rat kidney tissue labeling Heparan Sulfate Proteoglycan 2/Perlecan with ab318284 at 1/1000 (0.486 ug/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Positive staining on basement membrane of rat kidney (PMID: 24035513).
The section was incubated with ab318284 for 30 mins at room temperature.
The immunostaining was performed on a Leica Biosystems BOND™ RX instrument

Counterstained with Hematoxylin.

Secondary antibody only control: Secondary antibody is a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Heat mediated antigen retrieval was performed with Tris-EDTA buffer (pH 9.0, Epitope Retrieval Solution2) for 20 mins
Immunocytochemistry/ Immunofluorescence - Anti-Heparan Sulfate Proteoglycan 2/Perlecan antibody [RM2059] - BSA and Azide free (ab318285)

This data was developed using ab318284, the same antibody clone in a different buffer formulation.

Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% TritonX-100 permeabilized HUVEC (human umbilical vein endothelial cell) cells labelling Heparan Sulfate Proteoglycan 2/Perlecan with ab318284 at 1/1000 (0.486 ug/ml) dilution, followed by ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed antibody at 1/1000 2 ug/ml dilution (Green).

Confocal image showing membranous and cytoplasmic staining in HUVEC cell line (shown in green). The counterstain was observed in magenta. Nuclear DNA was labelled with DAPI (shown in blue).
Low expression: SH-SY5Y.
Image was taken with a confocal microscope (Leica-Microsystems, TCS SP8).

ab195889 Anti-alpha Tubulin mouse monoclonal antibody - Microtubule Marker (Alexa Fluor® 594) was used to counterstain tubulin at 1/200 2.5ug/ml dilution (Magenta). The Nuclear counterstain was DAPI (Blue).

Secondary antibody only control: Secondary antibody is ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed at 1/1000 2 ug/ml dilution.
Immunocytochemistry/ Immunofluorescence - Anti-Heparan Sulfate Proteoglycan 2/Perlecan antibody [RM2059] - BSA and Azide free (ab318285)

This data was developed using ab318284, the same antibody clone in a different buffer formulation.

Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% TritonX-100 permeabilized MEF (mouse embryo fibroblast) cells labelling Heparan Sulfate Proteoglycan 2/Perlecan with ab318284 at 1/1000 (0.486 ug/ml) dilution, followed by ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed antibody at 1/1000 2 ug/ml dilution (Green).

Confocal image showing cytoplasmic and membranous staining in MEF cell line (shown in green). The counterstain was observed in magenta. Nuclear DNA was labelled with DAPI (shown in blue).
Low expression: Neuro-2a.
Image was taken with a confocal microscope (Leica-Microsystems, TCS SP8).

ab195889 Anti-alpha Tubulin mouse monoclonal antibody - Microtubule Marker (Alexa Fluor® 594) was used to counterstain tubulin at 1/200 2.5ug/ml dilution (Magenta). The Nuclear counterstain was DAPI (Blue).

Secondary antibody only control: Secondary antibody is ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed at 1/1000 2 ug/ml dilution.
Flow Cytometry (Intracellular) - Anti-Heparan Sulfate Proteoglycan 2/Perlecan antibody [RM2059] - BSA and Azide free (ab318285)

This data was developed using ab318284, the same antibody clone in a different buffer formulation.

Flow cytometric analysis of 4% paraformaldehyde fixed 90% methanol permeabilized SH-SY5Y (human neuroblastoma epithelial cell) (Left) / HUVEC (human umbilical vein endothelial cell) (Right) cells labelling Heparan Sulfate Proteoglycan 2/Perlecan with ab318284 at 1/500 dilution (0.1ug) / Magenta (Red) compared with a Rabbit monoclonal IgG (ab172730) (Black) isotype control and an unlabelled control (cells without incubation with primary antibody and secondary antibody) (Blue).

Goat Anti-Rabbit IgG (Alexa Fluor® 488, ab150081) at 1/5000 dilution was used as the secondary antibody.

Low expression: SH-SY5Y (PMID:25883214).
Flow Cytometry (Intracellular) - Anti-Heparan Sulfate Proteoglycan 2/Perlecan antibody [RM2059] - BSA and Azide free (ab318285)

This data was developed using ab318284, the same antibody clone in a different buffer formulation.

Flow cytometric analysis of 4% paraformaldehyde fixed 90% methanol permeabilized Neuro-2a (mouse neuroblastoma neuroblast) (Left) / MEF (mouse embryo fibroblast) (Right) cells labelling Heparan Sulfate Proteoglycan 2/Perlecan with ab318284 at 1/500 dilution (0.1ug) / Magenta (Red) compared with a Rabbit monoclonal IgG (ab172730) (Black) isotype control and an unlabelled control (cells without incubation with primary antibody and secondary antibody) (Blue).

Goat Anti-Rabbit IgG (Alexa Fluor® 488, ab150081) at 1/5000 dilution was used as the secondary antibody.

Low expression: Neuro-2a
Immunoprecipitation - Anti-Heparan Sulfate Proteoglycan 2/Perlecan antibody [RM2059] - BSA and Azide free (ab318285)

This data was developed using ab318284, the same antibody clone in a different buffer formulation.
Heparan Sulfate Proteoglycan 2/Perlecan was immunoprecipitated from 0.35 mg EA.hy926 (human somatic cell hybrid endothelial cell) whole cell lysate with ab318284 at 1/30 dilution (2ug in 0.35mg lysates). Western blot was performed on the immunoprecipitate using ab318284 at 1/1000 dilution. VeriBlot for IP secondary antibody(HRP)(ab131366) was used at 1/5000 dilution.
Lane 1: EA.hy926 (human somatic cell hybrid endothelial cell) whole cell lysate
Lane 2: ab318284 IP in EA.hy926 (human somatic cell hybrid endothelial cell) whole cell lysate
Lane 3: Rabbit monoclonal IgG (ab172730) instead of ab318284 in EA.hy926 whole cell lysate
Blocking and dilution buffer and concentration: 5% NFDM/TBST.
Exposure time: 76 seconds..
Anti-Heparan Sulfate Proteoglycan 2/Perlecan antibody [RM2059] - BSA and Azide free (ab318285)

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.

Click here to view the general protocols

Datasheet download

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概述

产品名称

描述

宿主

特异性

经测试应用

种属反应性

免疫原

阳性对照

常规说明

性能

形式

存放说明

存储溶液

无载体

纯度

克隆

克隆编号

同种型

研究领域

相关产品

Alternative Versions

Compatible Secondaries

Isotype control

Related Products

应用

The Abpromise guarantee

靶标

功能

组织特异性

疾病相关

序列相似性

翻译后修饰

细胞定位

数据库链接

别名

图片

实验方案

数据表及文件

Datasheet download

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