Human Beta-Endorphin ELISA Kit (DEIA3206)

Regulatory status: For research use only, not for use in diagnostic procedures.

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Size
96T
Sample
serum, plasma, tissue homogenates and other biological fluids
Species Reactivity
Human
Intended Use
This kit is intended for quantitative detection of β-EP in serum, plasma, tissue homogenates and other biological fluids.
Storage
2-8°C for 6 months
Precision
Intra-Assay: CV<8%
Inter-Assay: CV<10%
General Description
Endorphins are endogenous opioid biochemical compounds. They are polypeptides produced by the pituitary gland and the hypothalamus in vertebrates, and they resemble the opiates in their abilities to produce analgesia and a sense of well-being. In other words, they might work as "natural pain killers." Using drugs may increase the effects of the endorphins. Beta-Endorphin is an endorphin produced by the pituitary gland that is a potent pain suppressant. Beta-endorphin is released into the blood (from the pituitary gland) and into the spinal cord and brain from hypothalamic neurons. The beta-endorphin that is released into the blood cannot enter the brain in large quantities because of the blood-brain barrier. The physiological importance of the beta-endorphin that can be measured in the blood is far from clear: beta-endorphin is a cleavage product of POMC which is the precursor hormone for adrenocorticotrophic hormone (ACTH), so it will be released whenever ACTH is released. The behavioural effects of beta-endorphin are exerted by its actions in the brain and spinal cord, and probably the hypothalamic neurons are the major source of beta-endorphin at these sites. In situations where the level of ACTH is increased (e.g. Addison disease), the level of endorphins also increases slightly.
Reconstitution And Storage
Bring all reagents and samples to room temperature for 20 minutes before use.
1. Wash Buffer
If crystals have formed in the concentrate, you can warm it with 40°C water bath (Heating temperature should not exceed 50°C) and mix it gently until the crystals have completely been dissolved. The solution should be cooled to room temperature before use.
Dilute 30ml (15ml for 48T) Concentrated Wash Buffer into 750ml (375ml for 48T) Wash Buffer with deionized or distilled water. Put unused solution back at 2-8°C.
2. Standards
1) Add 1 ml Sample Dilution Buffer into one Standard tube (labeled as zero tube), keep the tube at room temperature for 10 minutes and mix them thoroughly.
Note: If the standard tube concentration higher than the range of the kit,please dilute it and labeled as zero tube.
2) Label 7 EP tubes with 1/2, 1/4, 1/8, 1/16, 1/32, 1/64 and blank respectively. Add 0.3ml of the Sample Dilution Buffer into each tube. Add 0.3ml of the above Standard solution (from zero tube) into 1st tube and mix them thoroughly. Transfer 0.3ml from 1st tube to 2nd tube and mix them thoroughly. Transfer 0.3ml from 2nd tube to 3rd tube and mix them thoroughly, and so on. Sample Dilution Buffer was used for the blank control.

Note: It is best to use Standard Solutions within 2 hours.
3. Preparation of Biotin-labeled Antibody Working Solution
Prepare it within 1 hour before experiment.
1) Calculate required total volume of the working solution: 0.05ml/well × quantity of wells. (Allow 0.1-0.2ml more than the total volume.)
2) Dilute the Biotin-detection antibody with Antibody Dilution Buffer at 1:100 and mix them thoroughly. (i.e. Add 1μl Biotin-labeled antibody into 99μl Antibody Dilution Buffer.)
4. Preparation of HRP-Streptavidin Conjugate (SABC) Working Solution
Prepare it within 30 minutes before experiment.
1) Calculate required total volume of the working solution: 0.1ml/well × quantity of wells. (Allow 0.1-0.2ml more than the total volume.)
2) Dilute the SABC with SABC Dilution Buffer at 1:100 and mix them thoroughly. (i.e. Add 1μl of SABC into 99μl of SABC Dilution Buffer.)

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