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Index / Smart Serology / ErbB2 _ HER2 _ CD340 ELISA kit , Human ELISA kit /Product Detail : SEK10004 ErbB2 _ HER2 _ CD340 ELISA kit , Human ELISA kit
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ErbB2 _ HER2 _ CD340 ELISA kit , Human ELISA kit

 Price: 723   EUR
820   USD
561   GBP

Product name : ErbB2 _ HER2 _ CD340 ELISA kit , Human ELISA kit

Catalog number : SEK10004

Quantity: 5 Plates

Availability: Yes

Supplier name : Smart Serology

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About this Product :

ErbB2 _ HER2 _ CD340 ELISA kit , Human ELISA kit
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Description
ErbB2 _ HER2 _ CD340 ELISA kit , Human ELISA kit is the most suitable ELISA kit for your research - it is manufactured with the best quality antibodies and plates to provide you with high reproducible results in your lab work. The special designi of the buffers will guarantee optimal conditions in each phase: from dilution, through the incubation to the blocking and washing.

ErbB2 _ HER2 _ CD340 ELISA kit , Human ELISA kit Human samples 80 % of the research is conducted on human samples. Gentaur suppliers human normal cells, cell lines, RNA extracts and lots of antibodies and elisa kit to Human proteins as well as ErbB2 _ HER2 _ CD340 ELISA kit , Human ELISA kit.
http://mybiofast.com/ver.php?search=human+&submit=Search

More Details about

1. Materials provided

Capture Antibody - 0.5 mg/mL of mouse anti-ErbB2 monoclonal antibody. Dilute to a working concentration of  0.5 μg/mL in CBS before coating.

Detection Antibody - 0.28 mg/mL of biotinylated rabbit anti-ErbB2 polyclonal antibody. Dilute to a working concentration of 1.0 μg/mL in detection antibody dilution buffer before use.

Standard - Each vial contains 14ng of recombinant ErbB2. Reconstitute with 1 mL detection antibody dilution buffer. After reconstitution, store at -20℃ to -70℃ in a manual defrost freezer. A seven-point standard curve using 2-fold serial dilutions in sample dilution buffer, and a high standard of  1ng/mL  is recommended.

Streptavidin-HRP - 50 μL of streptavidin conjugated to horseradish-peroxidase. 1:2000  Dilution in detection antibody dilution buffer before use.

 

2. Sensitivity

The minimum detectable dose of human ErbB2 ( HER2 / CD340 ) was determined to be approximately 15.6 pg/ml. This is defined as at least three times standard deviations above the mean optical density of 10 replicates of the zero standard.

 

3. Principle of the product

The human ErbB2 ( HER2 / CD340 ) ELISA Pair Set is for the quantitative determination of human ErbB2.

This ELISA Pair Set contains the basic components required for the development of sandwich ELISAs. Each Set contains sufficient materials to run ELISAs on five 96-well plates.

The Sino Biological ELISA Pair Set is a solid phase sandwich ELISA (Enzyme-Linked Immunosorbent Assay). It utilizes a monoclonal antibody specific for ErbB2 ( HER2 / CD340 ) coated on a 96-well plate. Standards and samples are added to the wells, and any ErbB2 present binds to the immobilized antibody. The wells are washed and a biotinylated rabbit anti-ErbB2 ( HER2 / CD340 ) polyclonal antibody is then added, producing an antibody-antigen-antibody “sandwich”, which produces color in proportion to the amount of ErbB2 present in the sample strepavidin-HRP and TMB substrate solution are loaded. The absorbances of the microwell are read at 450 nm.

STORAGE - Keep streptavidin-HRP at 4℃ and protect it from prolonged exposure to light. Aliquot all other reagents and store at -20℃ to -70℃ in a manual defrost freezer.

ErbB2 / HER2 / CD340 ELISA PROTOCOL

 

1. Plate Preparation

  1. Dilute the capture antibody to the working concentration in CBS. Immediately coat a 96-well microplate with 100μL per well of the diluted capture antibody. Seal the plate and incubate overnight at 4℃.
  2. Aspirate each well and wash with at least 300μl wash buffer, repeating the process two times for a total of three washes. Complete removal of liquid at each step is 3. essential for good performance. After the last wash, remove any remaining wash buffer by inverting the plate and blotting it against clean paper towels.
  3. Block plates by adding 300 μL of blocking buffer to each well. Incubate at room temperature for a minimum of 1 hour.
  4. Repeat the aspiration/wash as in step 2. The plates are now ready for sample addition.

 

2. Assay Procedure

  1. Add 100 μL of sample or standards in sample dilution buffer per well. Seal the plate and incubate 2 hours at room temperature.
  2. Repeat the aspiration/wash as in step 2 of plate preparation.
  3. Add 100 μL of the detection antibody, diluted in antibody dilution buffer, to each well. Seal the plate and incubate 1 hour at room temperature.
  4. Repeat the aspiration/wash as in step 2 of plate preparation.
  5. Add 100 μL of Streptavidin-HRP to each well. Incubate for 1 hour at room temperature.
  6. Repeat the aspiration/wash as in step 2 of plate preparation.
  7. Add 200 μL of substrate solution to each well. Incubate for 20 minutes at room temperature ( if substrate solution is not as requested, the incubation time should be optimized ). Avoid placing the plate in direct light.
  8. Add 50 μL of stop solution to each well. Gently tap the plate to ensure thorough mixing.
  9. Determine the optical density of each well immediately, using a microplate reader set to 450 nm.

 

3. Calculation of results

  1. Calculate the mean absorbance for each set of duplicate standards, controls and samples. Subtract the mean zero standard absorbance from each.
  2. Construct a standard curve by plotting the mean absorbance for each standard on the y-axis against the concentration on the x-axis and draw a best fit curve through the points on the graph.
  3. To determine the concentration of the unknowns, find the unknowns’ mean absorbance value on the y-axis and draw a horizontal line to the standard curve. At the point of intersection, draw a vertical line to the x-axis and read the concentration. If samples have been diluted, the concentration read from the standard curve must be multiplied by the dilution factor.
  4. Alternatively, computer-based curve-fitting statistical software may also be employed to calculate the concentration of the sample.

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