Overview

  • Product name
  • Description
    Rabbit polyclonal to Hsp47
  • Tested applications
    Suitable for: IHC-Fr, WB, ICC/IF, IP, IHC-Pmore details
  • Species reactivity
    Reacts with: Mouse, Rat, Human
    Predicted to work with: Cow, Xenopus laevis, Non human primates
  • Immunogen

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

    (Peptide available as ab88169.)

  • Positive control
    • 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.

Properties

  • Form
    Liquid
  • Storage instructions
    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.
  • Storage buffer
    Preservative: 0.02% Sodium Azide
    Constituents: 1% BSA, PBS, pH 7.4
  • Concentration information loading...
  • Purity
    Immunogen affinity purified
  • Clonality
    Polyclonal
  • Isotype
    IgG
  • Research areas

Applications

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.

Application Abreviews Notes
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.

Target

  • Function
    Binds specifically to collagen. Could be involved as a chaperone in the biosynthetic pathway of collagen.
  • Involvement in disease
    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.
  • Sequence similarities
    Belongs to the serpin family.
  • Cellular localization
    Endoplasmic reticulum lumen.
  • Information by UniProt
  • Database links
  • Alternative names
    • 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 images

  • 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

References for Anti-Hsp47 antibody (ab77609)

This product has been referenced in:
  • Mimura H  et al. Functional Role of HSP47 in the Periodontal Ligament Subjected to Occlusal Overload in Mice. Int J Med Sci 13:248-54 (2016). IHC . Read more (PubMed: 27076780) »
  • 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) »

See all 4 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...

Read More

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...

Read More
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
Username

Abcam user community

Verified customer

Submitted Nov 08 2010

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

Sign up