CSF2 Antibody (OASA07792)

Catalog#: OASA07792

• Size: 50ug

• Description of Target: RABBIT ANTI HUMAN GM-CSF:Biotin
• Gene Symbol: CSF2
• Alias Symbols: GMCSF; MGC131935; MGC138897; CSF2
• Protein Accession#: NP_000749.2
• Swissprot Id: P04141
• Host: Rabbit
• Protein Size (# AA): 144
• Specificity: GM-CSF
• Application: ELISA, WB
• Conjugation: Biotin
• Immunogen: The immunogen for anti-CSF2 antibody: recombinant human GM-CSF
• Product Format: Purified IgG conjugated to Biotin - lyophilised
• Isotype: Polyclonal IgG
• Predicted Homology Based on Immunogen Sequence: Human
• Applications Info: ELISA : This product may be used in a direct ELISA or as the detection reagent in a sandwich ELISA with OASA07791 as the capture antibody and OPSA11082 as the standard
  Preservative Stabilisers: None present
  
  Antiserum Preparation: Antisera to human GM-CSF were raised by repeated immunisations of rabbits with highly purified antigen. Purified IgG prepared by affinity chromatography.
  Approx Protein Conc: IgG concentration 0.1 mg/ml after reconstitution
  
  Buffer Solutions: Phosphate buffered saline
• Key Reference: 1. Metcalf, D. (1986) The molecular biology and functions of the Granulocyte-Macrophage colony-stimulating factors. Blood. 67: 257-267.
• Reconstitution and Storage: Reconstitution: Reconstitute with 0.5 ml sterile PBS containing 0.1% Bovine Serum Albumin.
  Care should be taken during reconstitution as the protein may appear as a film at the bottom of the vial. Aviva Systems Biology recommend that the vial is gentl

Product Protocol: CSF2 antibody used to evaluate granulocyte/macrophage-colony-stimulating factor autoantibodies and myeloid cell immune functions in healthy subjects (OASA07792)

Specificity: GM-CSF
Product Page: CSF2 antibody (OASA07792)
Antibody Pair Available: OASA07791 Capture OASA07792 Detection
Experiment Type: Total GM-CSF using Sandwich ELISA

Protocol:
Serum concentrations of total GM-CSF (ie, autoantibody-bound plus free GM-CSF) were measured using a novel sandwich ELISA based on a method developed for measurement of HIV-1 p24 antigen. Microtiter plates were coated with capture antibody (PBS containing 1 μg/mL rabbit anti–human GM-CSF antibody, 4°C, overnight), and blocked with PBS containing 1% (wt/vol) BSA. To evaluate the detection of free GM-CSF and autoantibody-bound GM-CSF in serum, a set of standard samples composed of recombinant human GM-CSF (Leukine) ranging from 0 to 30 ng/mL was prepared in mouse serum in the absence or presence of purified human GM-CSF autoantibodies (30 μg/mL). Aliquots (10 μL) of serum or standard control samples were mixed with 20 μL pretreatment buffer (SDS-HD buffer; PBS, 1% SDS, 1.5 mM diethylenetriamine pentaacetic acid, pH 7.4, and incubated (95°C, 5 minutes). After chilling briefly on ice, 270 μL PBS containing 1% (wt/vol) BSA was added to quench residual SDS. Duplicate aliquots (50 μL) of samples were applied to wells, and the plates were incubated (room temperature, 90 minutes), washed, and then incubated with detection antibody solution (PBS containing 1% [wt/vol] BSA, 500 ng/mL biotin-conjugated goat anti–human GM-CSF antibody; room temperature, 90 minutes). Detection was enhanced by incubation with ExtrAvidin HRP solution (diluted 1/250; room temperature, 30 minutes), imaged by TMB solution followed by addition of 1 N H2SO4 and read at a wavelength of 450 nm with Benchmark ELISA plate reader.

Summary:
1. GM-CSF-binding immunoglobulins similar in molecular weight to IgG were detected in all healthy subjects evaluated, in pharmaceutical IVIG, and in a patient with PAP. 2. Similar levels of total serum GM-CSF levels were observed in patients with PAP. In contrast, levels of free GM-CSF measured using the commercial ELISA were less than 1 pg/mL in both groups. 3. GM-CSF autoantibodies seem to scavenge free GM-CSF in vivo and modulate the endocrine signaling capacity of GM-CSF in whole blood, suggesting that they may function to negatively regulate GM-CSF signaling in health and disease.

References:
1: Uchida K, Nakata K, Suzuki T, Luisetti M, Watanabe M, Koch DE, Stevens CA, Beck DC, Denson LA, Carey BC, Keicho N, Krischer JP, Yamada Y, Trapnell BC. Granulocyte/macrophage-colony-stimulating factor autoantibodies and myeloid cell immune functions in healthy subjects. Blood. 2009 Mar 12;113(11):2547-56. PubMed PMID: 19282464; PubMed Central PMCID: PMC2656275.