Anti-SPARC antibody

Anti-SPARC antibody

• 概述

• 产品描述This gene encodes a cysteine-rich acidic matrix-associated protein. The encoded protein is required for the collagen in bone to become calcified but is also involved in extracellular matrix synthesis and promotion of changes to cell shape. The gene product has been associated with tumor suppression but has also been correlated with metastasis based on changes to cell shape which can promote tumor cell invasion. Three transcript variants encoding different isoforms have been found for this gene. Appears to regulate cell growth through interactions with the extracellular matrix and cytokines. Binds calcium and copper, several types of collagen, albumin, thrombospondin, PDGF and cell membranes. There are two calcium binding sites; an acidic domain that binds 5 to 8 Ca(2+) with a low affinity and an EF-hand loop that binds a Ca(2+) ion with a high affinity.
• 产品名称Anti-SPARC antibody
• 分子量33 KDa
• 种属反应性Human,Mouse,Rat,Dog,Pig,Cow,Horse,Rabbit
• 验证应用WB,IHC-P,ICC/IF
• 抗体类型兔多抗
• 免疫原KLH conjugated synthetic peptide derived from human SPARC 101-200/303
• 偶联Non-conjugated

• 性能

• 形态Liquid
• 浓度1 mg/mL.
• 存放说明Store at -20℃ for one year. Avoid repeated freeze/thaw cycles. The lyophilized antibody is stable at room temperature for at least one month and for greater than a year when kept at -20℃. When reconstituted in sterile pH 7.4 0.01M PBS or diluent of antibody the antibody is stable for at least two weeks at 2-4℃.
• 存储缓冲液0.01M TBS(pH7.4) with 1% BSA, 0.03% Proclin300 and 50% Glycerol.
• 亚型IgG
• 纯化方式affinity purified by Protein A
• 亚细胞定位Secreted, extracellular space, extracellular matrix, basement membrane. Note=In or around the basement membrane.
• 其它名称AA517111
  Basement membrane protein 40
  BM 40
  BM40
  Cysteine rich protein
  hm:zeh0062
  MGC128090
  ON
  Osteonectin
  Secreted acidic cystein rich glycoprotein
  Secreted protein acidic and rich in cysteine
  Secreted protein acidic cysteine rich (osteonectin)
  Secreted protein acidic cysteine rich
  SPRC
  SPRC_HUMAN.
  more

• 应用

  WB:1:500-2000
  IHC-P:1:400-800
  IF:1:100-500

•  

  

  Fig1: Sample:
  Brain (Mouse) Lysate at 40 ug
  Primary: Anti-SPARC at 1/300 dilution
  Secondary: IRDye800CW Goat Anti-Rabbit IgG at 1/20000 dilution
  Predicted band size: 33 kD
  Observed band size: 33 kD

   

  

  Fig2: Tissue/cell: Human Meningioma; 4% Paraformaldehyde-fixed and paraffin-embedded;
  Antigen retrieval: citrate buffer ( 0.01M, pH 6.0 ), Boiling bathing for 15min; Block endogenous peroxidase by 3% Hydrogen peroxide for 30min; Blocking buffer (normal goat serum,C-0005) at 37℃ for 20 min;
  Incubation: Anti-SPARC Polyclonal Antibody, Unconjugate1:200, overnight at 4℃, followed by conjugation to the secondary antibody(SP-0023) and DAB(C-0010) staining

   

  

  Fig3: Tissue/cell: Human colon cancer; 4% Paraformaldehyde-fixed and paraffin-embedded;
  Antigen retrieval: citrate buffer ( 0.01M, pH 6.0 ), Boiling bathing for 15min; Block endogenous peroxidase by 3% Hydrogen peroxide for 30min; Blocking buffer (normal goat serum,C-0005) at 37℃ for 20 min;
  Incubation: Anti-SPARC Polyclonal Antibody, Unconjugate 1:200, overnight at 4℃, followed by conjugation to the secondary antibody(SP-0023) and DAB(C-0010) staining

   

  

  Fig4: Tissue/cell: rat brain tissue;4% Paraformaldehyde-fixed and paraffin-embedded;
  Antigen retrieval: citrate buffer ( 0.01M, pH 6.0 ), Boiling bathing for 15min; Blocking buffer (normal goat serum,C-0005) at 37℃ for 20 min;
  Incubation: Anti-SPARC Polyclonal Antibody, Unconjugated(1:200, overnight at 4℃; The secondary antibody was Goat Anti-Rabbit IgG, Cy3 conjugated(bs-0295G-Cy3)used at 1:200 dilution for 40 minutes at 37℃. DAPI(5ug/ml,blue,C-0033) was used to stain the cell nuclei

   

  

  Fig5: Tissue/cell: mouse kidney tissue;4% Paraformaldehyde-fixed and paraffin-embedded;
  Antigen retrieval: citrate buffer ( 0.01M, pH 6.0 ), Boiling bathing for 15min; Blocking buffer (normal goat serum,C-0005) at 37℃ for 20 min;
  Incubation: Anti-SPARC Polyclonal Antibody, Unconjugated(1:200, overnight at 4℃; The secondary antibody was Goat Anti-Rabbit IgG, Cy3 conjugated(bs-0295G-Cy3)used at 1:200 dilution for 40 minutes at 37℃. DAPI(5ug/ml,blue,C-0033) was used to stain the cell nuclei

   

  

  Fig6: Sample:
  A549(Human) Cell Lysate at 30 ug
  Primary: Anti-SPARC  at 1/1000 dilution
  Secondary: IRDye800CW Goat Anti-Rabbit IgG at 1/20000 dilution
  Predicted band size: 33 kD
  Observed band size: 35 kD

   

  

  Fig7: Paraformaldehyde-fixed, paraffin embedded (human brain glioma); Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15min; Block endogenous peroxidase by 3% hydrogen peroxide for 20 minutes; Blocking buffer (normal goat serum) at 37℃ f

   

  

  Fig8: Paraformaldehyde-fixed, paraffin embedded (mouse brain tissue); Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15min; Block endogenous peroxidase by 3% hydrogen peroxide for 20 minutes; Blocking buffer (normal goat serum) at 37℃ f

   

  

  Fig9: Paraformaldehyde-fixed, paraffin embedded (Rat brain); Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15min; Block endogenous peroxidase by 3% hydrogen peroxide for 20 minutes; Blocking buffer (normal goat serum) at 37℃ for 30min;

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