Anti-Collagen II antibody (bs-10589R)

Anti-Collagen II antibody

DATASHEET

Host:Rabbit

Target Protein:Collagen II

IR:Immunogen Range:231-330/1487

Clonality:Polyclonal

Isotype:IgG

Entrez Gene:1280

Swiss Prot:P02458

Source:KLH conjugated synthetic peptide derived from human Collagen II:231-330/1487 

Purification:affinity purified by Protein A

Storage:0.01M TBS(pH7.4) with 1% BSA, 0.03% Proclin300 and 50% Glycerol. Shipped at 4℃. Store at -20 °C for one year. Avoid repeated freeze/thaw cycles.

Background:Collagens are highly conserved throughout evolution and are characterized by an uninterrupted "Glycine-X-Y" triplet repeat that is a necessary part of the triple helical structure. For these reasons it is often extremely difficult to generate antibodies with specificities to collagens. The development of type specific antibodies is dependent on NON DENATURED three dimensional epitopes. This may result in diminished reactivity of some antibodies with denatured collagen or formalin fixed, paraffin embedded tissues. Type II collagen is a fibrillar collagen found in cartilage and the vitreous humor of the eye. Collagen type II is essential for the normal embryonic development of the skeleton, for linear growth and for the ability of cartilage to resist compressive forces. Mutations in this gene are associated with achondrogenesis, chondrodysplasia, early onset familial osteoarthritis, SED congenita, Langer Saldino achondrogenesis, Kniest dysplasia, Stickler syndrome type I, and spondyloepimetaphyseal dysplasia Strudwick type. In addition, defects in processing chondrocalcin, a calcium binding protein that is the C propeptide of this collagen molecule, are also associated with chondrodysplasia. There are two transcripts identified for this gene.

Size:100ul

Concentration:1mg/ml

Applications:WB(1:500-2000)
ELISA(1:5000-10000)
IHC-P(1:100-500)
IHC-F(1:100-500)
ICC(1:100-500)
IF(1:100-500)

Cross Reactive Species:Human
Mouse
Chicken
Dog
Pig
Cow
Horse
Rabbit
.

For research use only. Not intended for diagnostic or therapeutic use.

VALIDATION IMAGES

Sample:
293T(Human) Cell Lysate at 30 ug
A549 (Human) Cell Lysate at 30 ug
LOVO (Human) Cell Lysate at 40 ug
MCF-7 (Human) Cell Lysate at 30 ug
HepG2 (Human) Cell Lysate at 30 ug
Primary: Anti-Collagen II (bs-10589R) at 1/300 dilution
Secondary: IRDye800CW Goat Anti-Rabbit IgG at 1/20000 dilution
Predicted band size: 117 kD
Observed band size: 117 kD

Sample: bone(Mouse) Lysate at 40 ug
Primary: Anti-Collagen II(bs-10589R) at 1/300 dilution
Secondary: IRDye800CW Goat Anti-Rabbit IgG at 1/20000 dilution
Predicted band size: 117 kD
Observed band size: 134 kD

Sample:
MG63 (Human) Cell Lysate at 30 ug
Primary: Anti-Collagen II (bs-10589R) at 1/300 dilution
Secondary: IRDye800CW Goat Anti-Rabbit IgG at 1/20000 dilution
Predicted band size: 117 kD
Observed band size: 117 kD

Paraformaldehyde-fixed, paraffin embedded (mouse ear); 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°C for 30min; Antibody incubation with (Collagen II) Polyclonal Antibody, Unconjugated (bs-10589R) at 1:400 overnight at 4°C, followed by a conjugated secondary (sp-0023) for 20 minutes and DAB staining.

Tissue/cell: Mouse embryo tissue; 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-Collagen II Polyclonal Antibody, Unconjugated(bs-10589R) 1:100, overnight at 4∑C, followed by conjugation to the secondary antibody(SP-0023) and DAB(C-0010) staining

Paraformaldehyde-fixed, paraffin embedded (Rat lung); 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°C for 30min; Incubation: Anti-Collagen II Antibody, conjugated (bs-10589R-AF488) at 1:200 overnight at 4°C; DAPI (5ug/ml, blue, C-0033) was used to stain the cell nuclei.

Paraformaldehyde-fixed, paraffin embedded (Mouse lung); 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°C for 30min; Incubation: Anti-Collagen II Antibody, conjugated (bs-10589R-AF488) 1:400, 1.5 hours at 37°C; DAPI (5ug/ml, blue, C-0033) was used to stain the cell nuclei.