Collagen II (COL2A1) Rabbit Polyclonal Antibody

Specifications

Product Data
Applications	ELISA, IHC, IP, WB
Recommend Dilution	Anti-Collagen antibodies have been used for Indirect trapping ELISA for quantitation of antigen in serum using a standard curve, for immunoprecipitation and for native (non-denaturing, non-dissociating) PAGE and Western blotting for highly sensitive qualitative analysis. Recommended Dilutions: ELISA: 1/5,000-1/50,000. Western Blot: 1/1,000-1/10,000. Immunoprecipitation: 1/100. Immunohistochemistry: 1/50-1/400. QC: This product was assayed by Immunoblot and found to be reactive against Collagen II at a dilution of 1/5,000 to 1/10,000. This product was also assayed against 1.0 µg of Collagen II in a standard Sandwich ELISA using Peroxidase conjugated affinity purified anti-Rabbit IgG [H&L] (Goat) and (ABTS (2,2'-azino-bis-[3-ethylbenthiazoline-6-sulfonic acid]) as a substrate for 30 minutes at room temperature. A working dilution of 1/4,000-1/8,000 of the stock concentration showed best results.
Reactivity	Bovine, Human, Mouse, Rat, Sheep
Host	Rabbit
Clonality	Polyclonal
Immunogen	Collagen type II purified from Human knee and Bovine nasal cartilage.
Specificity	This product has been prepared by Immunoaffinity Chromatography using immobilised antigens followed by extensive cross-adsorbtion against other collagens, Human serum proteins and non-collagen extracellular matrix proteins to remove any unwanted specificities. Crossreactivity < 1% against other types of collagens was detected by ELISA against purified standards. Some class specific anti-collagens may be specific for three-dimensional epitopes which may result in diminished reactivity with denatured collagen or formalin-fixed, paraffin embedded tissues. This antibody reacts with most mammalian Type II collagens and has negligible cross-reactivity with Type I, III, IV, V or VI collagens. Non-specific cross reaction of anti-collagen antibodies with other human serum proteins or non-collagen extracellular matrix proteins is negligible.
Formulation	0.02M Sodium Phosphate, 0.15M Sodium Chloride, pH 7.2 State: Purified State: Liquid (sterile filtered) purified Ig fraction Stabilizer: None Preservative: 0.01% (w/v) Sodium Azide
Concentration	lot specific
Purification	Immunoaffinity Chromatography
Conjugation	Unconjugated
Storage Condition	Store undiluted at 2-8°C for one month or (in aliquots) at -20°C for longer. For extended storage, mix with an equal volume of glycerol. Avoid repeated freezing and thawing.
Gene Name	collagen type II alpha 1 chain
Database Link	Entrez Gene 1280 Human UniProt ID P02458
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. For immunization collagens from human and bovine placenta and cartilage extensively purified by limited pepsin digestion and selective salt precipitation were used. This preparation results in a native conformation of the protein. Antibodies are isolated from rabbit antiserum and are extensively cross-adsorbed by immunoaffinity purification to produce 'type' specific antibodies. Greatly diminished reactivity and selectivity of these antibodies will result if denaturing and reducing conditions of SDS-PAGE and immunoblotting are used.
Synonyms	COL2A1, Alpha-1 type II collagen
Reference Data

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