Albumin ELISA 试剂盒

1. Anti-HSA Antibody Coated Plate : One 96-well strip plate (8 x 12).
2. Biotinylated Anti-HSA Antibody (1000X) : One 20 μL vial.
3. Streptavidin-Enzyme Conjugate : One 20 μL vial.
4. Assay Diluent : One 50 mL bottle.
5. 10X Wash Buffer : One 100 mL bottle.
6. Substrate Solution : One 12 mL amber bottle.
7. Stop Solution (Part. No. 310808): One 12 mL bottle.

Box 2 (shipped on blue ice packs)

1. Plasma, Serum, Urine or Other Biological Fluids
2. PBS containing 0.1 % BSA
3. 10 μL to 1000 μL adjustable single channel micropipettes with disposable tips
4. 50 μL to 300 μL adjustable multichannel micropipette with disposable tips
5. Multichannel micropipette reservoir
6. Microplate reader capable of reading at 450 nm (620 nm as optional reference wave length)

• Sensitivity limit of 100 pg/mL human albumin
• Detection and quantitation of human albumin in plasma, serum, urine or other biological fluid samples

1X Wash Buffer: Dilute the 10X Wash Buffer Concentrate to 1X with deionized water. Stir to homogeneity. Biotinylated Anti-HSA Antibody and Streptavidin-Enzyme Conjugate: Immediately before use dilute the Biotinylated Anti-HSA antibody 1:1000 and the Streptavidin-Enzyme Conjugate 1:2000 with Assay Diluent. Do not store diluted solutions.

The following recommendations are only guidelines and may be altered to optimize or complement the user's experimental design. Plasma: Collect blood with heparin or EDTA and centrifuge at 4 °C for 10 minutes at 1000 g. Remove the plasma and assay immediately or store samples at -80 °C for up to three months. Normal plasma samples require about 1,000,000 fold dilution with PBS containing 0.1 % BSA immediately before running the ELISA. Prepare a serial dilution of each sample according to Table 2 below, mixing each dilution well before continuing to the next dilution step. Serum: Harvest serum and centrifuge at 4 °C for 10 minutes at 1000 g. Assay immediately or store samples at -80 °C for up to three months. Normal serum samples require about 1,000,000 fold dilution with PBS containing 0.1 % BSA immediately before running the ELISA. Prepare a serial dilution of each sample according to Table 2 below, mixing each dilution well before continuing to the next dilution step. Sample Plasma or Serum PBS with Effective Tubes Sample 0.1 % BSA Dilution 1 5 μL 5 mL 1:1000 2 5 μL of Tube #1 5 mL 1:1,000,000 Table 2. Preparation of 1:1,000,000 dilution of plasma or serum samples. Urine: Harvest urine and centrifuge for 10 minutes at 1000 g at 4 °C. Assay immediately or store samples at -80 °C for up to three months. Normal urine samples require about 1000 fold dilution with PBS containing 0.1 % BSA immediately before running the ELISA. Other Biological Fluids: Centrifuge samples for 10 minutes at 1000 g at 4 °C. Assay immediately or store samples at -80 °C for up to three months.

1. Prepare dilutions of plasma, serum, urine or other biological fluid samples in PBS containing 0.1 % BSA.
2. Add 100 μL of human albumin unknown sample or standard to the Anti-HSA Antibody Coated Plate. Each human albumin unknown sample, standard and blank should be assayed in duplicate.
3. Incubate at 37 °C for at least 2 hours or 4 °C overnight.
4. Wash microwell strips 3 times with 250 μL 1X Wash Buffer per well with thorough aspiration between each wash. After the last wash, empty wells and tap microwell strips on absorbent pad or paper towel to remove excess 1X Wash Buffer.
5. Add 100 μL of the diluted Biotinylated Anti-HSA antibody to each well. Incubate at room temperature for 1 hour on an orbital shaker. 4
6. Wash the strip wells 3 times according to step 4 above.
7. Add 100 μL of the diluted Streptavidin-Enzyme Conjugate to each well. Incubate at room temperature for 1 hour on an orbital shaker.
8. Wash the strip wells 3 times according to step 4 above. Proceed immediately to the next step.
9. Warm Substrate Solution to room temperature. Add 100 L of Substrate Solution to each well, including the blank wells. Incubate at room temperature on an orbital shaker. Actual incubation time may vary from 2-30 minutes. Note: Watch plate carefully, if color changes rapidly, the reaction may need to be stopped sooner to prevent saturation.
10. Stop the enzyme reaction by adding 100 μL of Stop Solution into each well, including the blank wells. Results should be read immediately (color will fade over time).
11. Read absorbance of each microwell on a spectrophotometer using 450 nm as the primary wave length. 5