SBA Clonotyping System-AP Kit (OKSB00002)

• Product Number: OKSB00002
• Name: SBA Clonotyping System-AP Kit (OKSB00002)
• Quality Control: To ensure lot to lot consistency, each batch of product is tested by ELISA for conformance to characteristics of a standard reference reagent.
• Host: Goat
• Clonality: Polyclonal
• Application Kit: Alkaline phosphatase (AP)- based Enzyme- Linked- Immunosorbent- Assay (ELISA)and immunoblotting
  - 1 mL goat anti- mouse Ig capture antibody (2.5 mg/mL)
  - 1 mL AP- labeled goat anti- mouse Ig screening antibody
  - 1 mL AP- labeled goat anti- mouse IgM
  - 1 mL AP- labeled goat anti- mouse IgG1
  - 1 mL AP- labeled goat anti- mouse IgG2a
  - 1 mL AP- labeled goat anti- mouse IgG2b
  - 1 mL AP- labeled goat anti- mouse IgG3
  - 1 mL AP- labeled goat anti- mouse IgA
  - 1 mL AP- labeled goat anti- mouse ?
  - 1 mL AP- labeled goat anti- mouse ?
  - p- nitrophenyl phosphate substrate tablets (20 x 5 mg)
• Reconstitution and Storage: Store at 2-8C
• Storage: - The purified (UNLB) antibody is supplied as 2.5 mg purified immunoglobulin in 1.0 mL of borate buffered saline, pH 8.2. No preservatives or amine containing buffer salts added. Store at 2-8C.
  The alkaline phosphatase (AP) conjugates are supplied as 1.0 mL of stock solution in 50mM Tris/1mM MgCl2/50%Glycerol, pH 8.0, containing 0.1% NaN3 as preservative. Store at 2-8C or long term at -20C.
  The pNPP substrate tablets are supplied as 20 x 5 mg. Recommended storage is -20C. Protect from light.
  Reagents are stable for the period shown on the label if stored as directed.
• Datasheets/Manuals: Click here to download product manual. As variation between lots may occur, always reference the lot-specific manual received with each kit.
• Application Info: Initial screening of fusions and selection of antibody-producing hybridomas, Identification of the heavy and light chain isotypes of antibodies, Selection and isolation of isotype switch variants
• Warning: Reagents contain sodium azide which is very toxic if ingested or inhaled. Avoid contact with skin, eyes, orclothing. Wear eye or face protection when handling. If skin or eye contact occurs, wash with copious amounts ofwater. If ingested or inhaled, contact a physician immediately. Sodium azide yields toxic hydrazoic acid underacidic conditions. Dilute azide- containing compounds in running water before discarding to avoid accumulation ofpotentially explosive deposits in lead or copper plumbing.
• Procedure: 1. Dilute capture antibody to a concentration of 5 - 10 ug/mL in borate buffered saline (BBS), pH 8.2 or phosphate buffered saline (PBS), pH 7.4; add 0.1 mL to each well of the ELISA plate; alternatively, the antigen used for immunization may be used as the coating reagent. 2. Cover plate with a lid or plastic wrap and incubate in a humidified atmosphere at 2-8C for a minimum of 12 hours. 3. Empty wells, wash 3X with BBS (or PBS) containing 0.05% Tween, empty wells, and fill wells with BBS (or PBS) containing 1% bovine serum albumin (BBS/BSA). 4. Allow antibody-coated plate to stand at room temperature for a minimum of 1 hour to block free binding sites on the plate. 5.Empty plate and wash 3X. 6. Add 0.1 mL of hybridoma supernatant to each well, cover plate, and incubate for 1 hour at room temperature with gentle shaking overnight at 2-8C. 7. Empty plate and wash 3X. 8. Dilute AP-labeled detection antibody(ies) 1:250-1:500 in BBS/BSA, add 0.1 mL conjugate(s) to appropriate wells of the plate, cover plate, and incubate for 1 hour at room temperature with gentle shaking or overnight at 2-8C. 9. Empty the plate and wash 5X. 10. Prepare substrate buffer - To 400 mL of double glass-distilled water, add 24.5 mg MgCl2-6H2O and 48 mL diethanolamine; adjust pH. to 9.8 with 5N HCl and make up to 500 mL with distilled water. 11. Prepare a 1 mg/mL substrate solution (e.g., one 5 mg tablet + 5 mL substrate buffer) and add 0.1 mL to each well of the plate. 12. Read optical density of each well at 405 nm after substrate addition.
• Tested Species Reactivity: Mouse
• Application: ELISA, ELISPOT