MOUSE ANTI CAT IgA:Biotin Antibody (0.25mg)

Catalog#: OASA00453

• Size: 0.25mg

• Description of Target: MOUSE ANTI CAT IgA:Biotin
• Alias Symbols: N/A
• Host: Mouse
• Protein Size (# AA): 131
• Specificity: IgA
• Application: ELISA
• Clone: CDA2-43A1
• Conjugation: Biotin
• Product Format: Purified IgG - liquid
• Isotype: IgG2a
• Applications Info: ELISA : This reagent may be used as a detection reagent in a sandwich ELISA system in combination with OASA041781 as capture reagent.
  Preparation: Purified IgG prepared by affinity chromatography on Protein A from tissue culture supernatant
  
  Preservative Stabilisers: 0.09% - Sodium Azide
  Approx Protein Conc: IgG concentration 0.5mg/ml
  
  Buffer Solutions: Phosphate buffered saline pH7.2
• Reconstitution and Storage: Store at +4^oC or at -20^oC if preferred.
  
  This product should be stored undiluted. Avoid repeated freezing and thawing as this may denature the antibody. Should this product contain a precipitate we recommend microcentrifugation before use.

Product Protocol: MOUSE ANTI CAT IgA:Biotin antibody used to evaluate innate imprinting by the modified heat-labile toxin of escherichia coli (ltk63) provides generic protection against lung infectious disease (OASA00453)

Specificity: IgA
Product Page: MOUSE ANTI CAT IgA:Biotin antibody (OASA00453)
Antibody Pair Available: OASA00451 Capture OASA00453 Detection
Experiment Type: IgA-specific Ab ELISA

Protocol:
Immuno Maxisorb ELISA plates (Nunc, Roskilde, Denmark) were coated with either sonicated RSV Ag or UV-inactivated influenza virus (100 HA/well) in PBS and incubated at 4°C overnight. Plates were washed with PBS-Tween (0.05%) and between each subsequent step. To each well, 200 μl of 10% BSA-PBS were added and incubated at room temperature for 90 min. BAL fluid was diluted 1/2 in 10% BSA-PBS, and each sample was incubated in duplicate for 2 h at room temperature. Then 100 μl of biotinylated anti-IgA (Serotec, Oxford, U.K.) diluted 1/500 were added to each well and incubated for 1 h at room temperature. To each well, 100 μl of streptavidin-HRP (DakoCytomation, Carpinteria, CA) diluted 1/500 were added and incubated for 1 h at room temperature. Finally 100 μl of substrate o-phenylenediamine (Sigma-Aldrich) solution were added to each well and incubated for 30 min; the reaction was stopped with 2 M H2SO4. Plates were read at 490 nm, and optical density was compared between sample groups (no standard was available for quantification).

Summary:
1. Treatment of mice with 5 μg of LTK63 14 days before influenza infection also reduced the proportion and total number of CD4+ T cells producing IFN-γ and IL-5. Despite reduced cytokine production, significant RSV- or influenza-specific IgA was detected in nasal washes taken from mice infected with G/RSV or influenza virus. In both cases, prior intranasal LTK63 treatment enhanced recovered Ag-specific IgA. 2. LTK63 treatment effectively matures the APC compartment of the respiratory tract and induces potent CD8+ T cells and local IgA production. Excessive inflammation is eliminated without sacrificing effective pathogen clearance, and it is most effective at the site of pathogen entry and replication.

References:
1: Williams AE, Edwards L, Humphreys IR, Snelgrove R, Rae A, Rappuoli R, Hussell T. Innate imprinting by the modified heat-labile toxin of Escherichia coli (LTK63) provides generic protection against lung infectious disease. J Immunol. 2004 Dec 15;173(12):7435-43. PubMed PMID: 15585869.