Fibronectin antibody (Biotin)

Catalog No. ABIN5596762

Product Details anti-Fibronectin Antibody (Biotin)

Target: Fibronectin

Reactivity: Human, Mouse, Rat, Cow, Monkey

Host: Rabbit

Clonality: Polyclonal

Conjugate: This Fibronectin antibody is conjugated to Biotin

Application: Western Blotting (WB), Immunohistochemistry (IHC), Immunoprecipitation (IP), ELISA

Cross-Reactivity (Details): Typically less than 1% cross reactivity against other extracellular matrix proteins was detected by ELISA against purified standards. This antibody reacts with human Fibronectin and has negligible cross-reactivity with Type I, II, III, IV, V or VI Collagens or Laminin. Non-specific cross-reaction of anti-Fibronectin antibodies with other human serum proteins or non-Fibronectin extracellular matrix proteins is negligible.

Purification: Anti-fibronectin (rabbit) antibody has been prepared by immunoaffinity chromatography using immobilized antigens followed by extensive cross-adsorption against human serum proteins and collagen and non-collagen extracellular matrix proteins to remove any unwanted specificity.

Immunogen:

Immunogen: Fibronectin was purified from Human plasma by binding to a denatured gelatin column followed by elution with high concentrations of arginine. The eluted material was further purified by gel filtration. Immunization occurred after single-band purity was assessed by SDS-PAGE.

Immunogen Type: Native Protein

Isotype: IgG

Labeling Ratio: 10-20

Product Specific Information:

This polyclonal rabbit anti-fibronectin antibody is biotin conjugated and is suitable for the detection of fibronectin (FN, FN1) in human, mouse, rat, bovine, monkey species. The Fibronectin antibody is validated for ELISA, immunohistochemistry, immunoprecipitation and western blotting. Four product images for IHC and unfolding profile are currently available as evidence of the antibody's performance. Use our high-quality Fibronectin antibody to reliably detect Fibronectin for a variety of species.

Fibronectins bind cell surfaces and various compounds including collagen, fibrin, heparin, DNA, and actin. Fibronectins are involved in cell adhesion, cell motility, opsonization, wound healing, and maintenance of cell shape. Involved in osteoblast compaction through the fibronectin fibrillogenesis cell-mediated matrix assembly process, essential for osteoblast mineralization (By similarity). Participates in the regulation of type I collagen deposition by osteoblasts (By similarity). (UniProt)

Application Details

Application Notes:

Immunohistochemistry Dilution: 1:50 - 1:200

Application Note: Anti-fibronectin (rabbit) antibody was assayed by immunoblot and found to be reactive against Fibronectin at a dilution of 1:5,000 to 1:10,000. This product was also assayed against 1.0 μg of Fibronectin in a standard capture ELISA using Peroxidase Conjugated Streptavidin #S000-03 and ABTS (2,2'-azino-bis-[3-ethylbenthiazoline-6-sulfonic acid]) code # ABTS-100 as a substrate for 30 minutes at room temperature. A working dilution of 1:4,000 to 1:8,000 of the stock concentration is suggested for this product. For immunohistochemistry on paraffin embedded tissue dilute the product 1:50 to 1:200.

Western Blot Dilution: 1:500 - 1:5,000

Immunoprecipitation Dilution: 1:100

ELISA Dilution: 1:5,000 - 1:20,000

Restrictions: For Research Use only

Independent Validation (UP)

Validation #104084 (Unfolding Profile)

by: NanoTemper Technologies

No.: #104084

Date: 07/23/2019

Antigen: Fibronectin

Lot Number: 39102

Method validated: Unfolding Profile

Positive Control: ABIN5596762

Notes:

Passed. ABIN5596762 showed T_i at 63.4°C and a clear unfolding profile with one unfolding event. This suggests that the antibody is properly folded and functional.

Validation Images

Unfolding profile of ABIN5596762. The fluorescence signal is plotted against temperature. The vertical line indicates the Ti at 63.4°C.

Full Methods

Full Protocol:

• Dilute ABIN5596762 in PBS buffer (Roth, 1058.1, lot 285231988) to get a final volume of 30µl at a concentration of 0.5µM.
• Load sample into Tycho capillary (NanoTemper Technologies, TY-C001).
• Run Tycho measurement.

Experimental Notes:

Tycho is designed to run quick and precise protein quality check experiments. Tycho uses intrinsic protein fluorescence to follow protein unfolding while running a fast thermal ramp, yielding results in 3min. A protein’s unfolding behavior is characterized by various parameters, most notably the inflection temperature (T_i). The T_i can be used to identify properly folded protein, to compare different batches, or to analyze the influence of storage/transport conditions on a protein. An absence of T_i would suggest that the protein is already unfolded and therefore most likely nonfunctional.

Handling

Format: Lyophilized

Reconstitution:

Reconstitution Volume: 100 μL

Reconstitution Buffer: Restore with deionized water (or equivalent)

Concentration: 1.0 mg/mL

Buffer:

Buffer: 0.02 M Potassium Phosphate, 0.15 M Sodium Chloride, pH 7.2

Stabilizer: 10 mg/mL Bovine Serum Albumin (BSA) - Immunoglobulin and Protease free

Preservative: Sodium azide

Precaution of Use: This product contains Sodium azide: a POISONOUS AND HAZARDOUS SUBSTANCE which should be handled by trained staff only.

Storage: 4 °C,-20 °C

Storage Comment: Store vial at 4° C prior to restoration. For extended storage aliquot contents and freeze at -20° C or below. Avoid cycles of freezing and thawing. Centrifuge product if not completely clear after standing at room temperature. This product is stable for several weeks at 4° C as an undiluted liquid. Dilute only prior to immediate use.

Expiry Date: 12 months

Target Details for Fibronectin

Target: Fibronectin

Abstract: Fibronectin Products

Synonyms: FN antibody, cig antibody, fibronectin antibody, finc antibody, lets antibody, msf antibody, CIG antibody, ED-B antibody, FINC antibody, FNZ antibody, GFND antibody, GFND2 antibody, LETS antibody, MSF antibody, E330027I09 antibody, Fn antibody, Fn-1 antibody, FIBNEC antibody, fn-1 antibody, fibronectin 1 antibody, FN1 antibody, fn1 antibody, Fn1 antibody

Background:

Synonyms: CIG antibody, Cold insoluble globulin antibody, LETS antibody, Migration stimulating factor antibody, MSF antibody, Transformation sensitive protein. Antibody

Background: Fibronectin antibody reacts with human fibronectin in liver, tonsil, skin and kidney. Traces of contaminating antibodies have been removed by solid-phase absorption. Biotin is ammenable to conjugation to proteins for use in biochemical assays. Biotin has a very strong affinity for avidin and streptavidin, an attraction that is the strongest and most stable non-covalent interaction known. Fibronectin is found in two forms in vertebrates: soluble and insoluble. Soluble plasma fibronectin is contained in blood plasma and constitutes a large protein component. Insoluble cellular fibronectin is a large component of the extra-cellular matrix where it is secreted by many different types of cells. Fibronectin plays a large role in wound healing and cell development. Anti-fibronectin (rabbit) antibody is ideal for investigators in Cardiology, Cell Biology, Microbiology, and Immunology research.

Gene Name: FN1

Gene ID: 2335

UniProt: P02751