概述

  • 产品名称Anti-Hsp47抗体
    参阅全部 Hsp47 一抗
  • 描述
    兔多克隆抗体to Hsp47
  • 经测试应用适用于: IHC-Fr, WB, ICC/IF, IP, IHC-Pmore details
  • 种属反应性
    与反应: Mouse, Rat, Human
    预测可用于: Cow, Xenopus laevis, Non Human Primates
  • 免疫原

    Synthetic peptide conjugated to KLH derived from within residues 350 to the C-terminus of Human Hsp47.

    (Peptide available as ab88169.)

  • 阳性对照
    • This antibody gave a positive signal in NIH3T3 whole cell lysate and in the following tissue lysates: Human Lung; Human Liver; Human Placenta; Rat Brain; Rat Placenta. It also gave a positive result in Human Skin. IHC-P: FFPE Human Skin Normal.

性能

  • 形式Liquid
  • 存放说明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.
  • 存储溶液Preservative: 0.02% Sodium Azide
    Constituents: 1% BSA, PBS, pH 7.4
  • Concentration information loading...
  • 纯度Immunogen affinity purified
  • 克隆多克隆
  • 同种型IgG
  • 研究领域

应用

Our Abpromise guarantee covers the use of ab77609 in the following tested applications.

The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.

应用 Ab评论 说明
IHC-Fr Use at an assay dependent concentration. PubMed: 24804164
WB Use a concentration of 1 µg/ml. Detects a band of approximately 48 kDa (predicted molecular weight: 46 kDa).
ICC/IF Use a concentration of 1 µg/ml.
IP Use a concentration of 5 µg/ml.
IHC-P Use a concentration of 1 - 5 µg/ml. Perform heat mediated antigen retrieval before commencing with IHC staining protocol.

靶标

  • 功能Binds specifically to collagen. Could be involved as a chaperone in the biosynthetic pathway of collagen.
  • 疾病相关Note=Defects in SERPINH1 may cause severe autosomal recessive osteogenesis imperfecta (OI). Osteogenesis imperfecta defines a group of connective tissue disorders characterized by bone fragility and low bone mass.
  • 序列相似性Belongs to the serpin family.
  • 细胞定位Endoplasmic reticulum lumen.
  • Information by UniProt
  • 数据库链接
  • 别名
    • 47 kDa heat shock protein antibody
    • 47 kDa heat shock protein precursor antibody
    • Arsenic transactivated protein 3 antibody
    • Arsenic-transactivated protein 3 antibody
    • AsTP 3 antibody
    • AsTP3 antibody
    • CBP 1 antibody
    • CBP 2 antibody
    • CBP1 antibody
    • CBP2 antibody
    • Cell proliferation-inducing gene 14 protein antibody
    • Collagen binding protein 1 antibody
    • Collagen binding protein 2 antibody
    • Collagen binding protein antibody
    • Collagen-binding protein 2 antibody
    • Collagen-binding protein antibody
    • Colligen antibody
    • Colligin 1 antibody
    • Colligin 2 antibody
    • Colligin antibody
    • colligin-1 antibody
    • colligin-2 antibody
    • gp46 antibody
    • Heat shock protein 47 antibody
    • Heat-shock protein 47 antibody
    • HGNC 1547 antibody
    • Hsp 47 antibody
    • J6 antibody
    • OI10 antibody
    • PIG 14 antibody
    • PIG14 antibody
    • PPROM antibody
    • Proliferation inducing gene 14 antibody
    • Proliferation inducing gene 14 protein antibody
    • RA A47 antibody
    • RA-A47 antibody
    • Rheumatoid arthritis antigen A 47 antibody
    • rheumatoid arthritis antigen A-47 antibody
    • Rheumatoid arthritis related antigen RA A47 antibody
    • Rheumatoid arthritis-related antigen RA-A47 antibody
    • serine (or cysteine) proteinase inhibitor, clade H (heat shock protein 47), member 1, (collagen binding protein 1) antibody
    • serine (or cysteine) proteinase inhibitor, clade H (heat shock protein 47), member 2, (collagen-binding protein 2) antibody
    • Serine or cysteine proteinase inhibitor clade H member 1 antibody
    • Serine or cysteine proteinase inhibitor clade H member 2 antibody
    • SERPH_HUMAN antibody
    • Serpin H1 antibody
    • Serpin peptidase inhibitor clade H member 1 antibody
    • serpin peptidase inhibitor, clade H (heat shock protein 47), member 1, (collagen binding protein 1) antibody
    • Serpin peptidase inhibitor, clade H, member 1 antibody
    • SERPINH1 antibody
    • SERPINH2 antibody
    see all

Anti-Hsp47 antibody 图像

  • IHC image of ab77609 staining in human skin 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 ab77609, 1µg/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.

    *Tissue obtained from the Human Research Tissue Bank, supported by the NIHR Cambridge Biomedical Research Centre.

  • All lanes : Anti-Hsp47 antibody (ab77609)

    Lane 1 : Lung (Human) Tissue Lysate
    Lane 2 : Human liver tissue lysate - total protein (ab29889)
    Lane 3 : Human placenta tissue lysate - total protein (ab29745)

    Lysates/proteins at 10 µg per lane.

    Secondary
    Goat polyclonal to Rabbit IgG - H&L - Pre-Adsorbed (HRP) at 1/3000 dilution
    Developed using the ECL technique

    Performed under reducing conditions.

    Predicted band size : 46 kDa
    Observed band size : 48 kDa (why is the actual band size different from the predicted?)
    Additional bands at : 52 kDa,85 kDa. We are unsure as to the identity of these extra bands.
  • ICC/IF image of ab77609 stained HeLa cells. The cells were 4% PFA 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 (ab77609, 1µg/ml) overnight at +4°C. The secondary antibody (green) was Alexa Fluor® 488 Goat anti-Rabbit IgG (H+L) used at a 1/1000 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 antibody also gave a positive result in 4% PFA fixed (10 min) HEK293, HepG2, and MCF-7 cells at 1µg/ml, and in 100% Methanol fixed (5 min) HeLa, HEK293, HepG2, and MCF-7 cells at 1µg/ml.
  • All lanes : Anti-Hsp47 antibody (ab77609) at 1 µg/ml

    Lane 1 : NIH 3T3 (Mouse embryonic fibroblast cell line) Whole Cell Lysate
    Lane 2 : Brain (Rat) Tissue Lysate
    Lane 3 : Spinal Cord (Rat) Tissue Lysate

    Lysates/proteins at 10 µg per lane.

    Secondary
    Goat Anti-Rabbit IgG H&L (HRP) preadsorbed (ab97080) at 1/5000 dilution
    Developed using the ECL technique

    Performed under reducing conditions.

    Predicted band size : 46 kDa
    Observed band size : 48 kDa (why is the actual band size different from the predicted?)
    Additional bands at : 100 kDa,55 kDa. We are unsure as to the identity of these extra bands.

    Exposure time : 1 minute

  • Developed using the ECL technique

    Performed under reducing conditions.

    Predicted band size : 46 kDa


    Exposure time : 10 seconds
  • Hsp47 was immunoprecipitated using 0.5mg NIH3T3 whole cell extract, 5µg of Rabbit polyclonal to Hsp47 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, NIH3T3 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 ab77609.
    Secondary: Mouse monoclonal [SB62a] Secondary Antibody to Rabbit IgG light chain (HRP) (ab99697).
    Band: 48kDa; Hsp47

Anti-Hsp47 antibody (ab77609)参考文献

This product has been referenced in:
  • Murayama MA  et al. CTRP6 is an endogenous complement regulator that can effectively treat induced arthritis. Nat Commun 6:8483 (2015). Read more (PubMed: 26404464) »
  • Kumar P  et al. MUC1 Is Expressed by Human Skin Fibroblasts and Plays a Role in Cell Adhesion and Migration. Biores Open Access 3:45-52 (2014). IHC-Fr, ICC/IF ; Human . Read more (PubMed: 24804164) »

See all 3 Publications for this product

Product Wall

Thank you for your reply.
The 5% BSA in 0.1% TBST solution is used for both the blocking bufferand the primary antibody diluent.

PVDF and nitrocellulose can usually be used interchangeably for most proteins. The primary differences betw...

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Thank you for contacting us.

This antibody has been validated using 5% BSA for blocking and a nitrocellulose membrane, but we have no reason to believe it would not work equally well using milk or PVDF. If you would like to optimize the proto...

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Abcam guarantees this product to work in the species/application used in this Abreview.
Application Immunocytochemistry/ Immunofluorescence
Sample Mouse Cell (mouse myoblasts)
Specification mouse myoblasts
Fixative Formaldehyde
Permeabilization Yes - TritonX-100 0.3%
Blocking step Serum as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 24°C
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提交于 Nov 08 2010

Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"