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Human Progesterone ELISA Kit

SKU:
HUFI00321
Product Type:
ELISA Kit
Size:
96 Assays
Uniprot:
Not available
Sensitivity:
0.188ng/ml
Range:
0.313-20ng/ml
ELISA Type:
Competitive
Synonyms:
PG, Progesterone
Reactivity:
Universal
$719
Frequently bought together:

Description

Human Progesterone ELISA Kit

Progesterone is a steroid hormone primarily produced by the ovaries in females and the adrenal glands in both genders. It plays a crucial role in the menstrual cycle, pregnancy, and maintaining the uterine lining for embryo implantation. Progesterone also helps regulate various physiological functions, including mood, metabolism, and immune response.

system_update_alt Datasheet system_update_alt MSDS

Key Features

Save Time Pre-coated 96 well plate
Quick Start Kit includes all necessary reagents
Publication Ready Reproducible and reliable results

Overview

Product Name:

Human Progesterone ELISA Kit

Product Code:

HUFI00321

Size:

96 Assays

Alias:

PG, Progesterone

Detection Method:

Competitive ELISA, Coated with Antibody

Application:

This immunoassay kit allows for the in vitro quantitative determination of PG concentrations in serum plasma and other biological fluids.

Sensitivity:

0.188ng/ml

Range:

0.313-20ng/ml

Storage:

4°C for 6 months

Note:

For Research Use Only

Additional Information

Recovery

Matrices listed below were spiked with certain level of PG and the recovery rates were calculated by comparing the measured value to the expected amount of PG in samples.

Matrix

Recovery Range (%)

Average (%)

serum(n=5)

85-105

97

EDTA plasma(n=5)

85-105

93

UFH plasma(n=5)

85-105

93

Linearity

The linearity of the kit was assayed by testing samples spiked with appropriate concentration of PG and their serial dilutions. The results were demonstrated by the percentage of calculated concentration to the expected.

Sample

1:2

1:4

1:8

serum(n=5)

88-103%

89-103%

90-105%

EDTA plasma(n=5)

83-100%

82-96%

82-101%

UFH plasma(n=5)

8-97%

80-95%

88-99%

CV(%)

Intra-Assay: CV<8%
Inter-Assay: CV<10%

Kit Components

Component Quantity Storage

ELISA Microplate (Dismountable)

8x12 strips

4°C for 6 months

Lyophilized Standard

2

4°C/ -20°C

Sample/Standard Dlution Buffer

20ml

4°C

Biotin-labeled Antibody (Concentrated)

120ul

4°C (Protection from light)

Antibody Dilution Buffer

10ml

4°C

HRP-Streptavidin Conjugate (SABC)

120ul

4°C (Protect from light)

SABC Dilution Buffer

10ml

4°C

TMB Substrate

10ml

4°C (Protection from light)

Stop Solution

10ml

4°C

Wash Buffer (25X)

30ml

4°C

Plate Sealer

5

-

Other materials required:

  • Microplate reader with 450 nm wavelength filter
  • Multichannel Pipette, Pipette, microcentrifuge tubes and disposable pipette tips
  • Incubator
  • Deionized or distilled water
  • Absorbent paper
  • Buffer resevoir

Protocol

*Note: Protocols are specific to each batch/lot. For the correct instructions please follow the protocol included in your kit.

Equilibrate the TMB substrate for at least 30 min at 37°C beforeuse. When diluting samples and reagents, they must be mixed completely andevenly. It is recommended to plot a standard curve for each test.

Step Procedure

1.

Set standard, test sample and control (zero) wells on the pre-coatedplate respectively, and then, record their positions. It isrecommended to measure each standard and sample in duplicate. Washplate 2 times before adding standard, sample and control (zero) wells!

2.

Add Sample and Biotin-detection antibody: Add 50µL of Standard, Blank or Sample per well. The blankwell is added with Sample Dilution Buffer. Immediately add 50 µL of biotin-labelled antibody workingsolution to each well. Cover with the plate sealer provided. Gently tap the plate to ensure thoroughmixing. Incubate for 45 minutes at 37°C. (Solutions are added to the bottom of micro-ELISA platewell, avoid touching plate walls and foaming).

3.

Wash: Aspirate each well and wash, repeating the process three timesWash by filling each well with Wash Buffer (approximately 350µL)using a squirt bottle, multi-channel pipette, manifold dispenser orautomated washer. Complete removal of liquid at each step is essentialto good performance. After the last wash, remove any remaining WashBuffer by aspirating or decanting. Invert the plate and pat it againstthick clean absorbent paper.

4.

HRP-Streptavidin Conjugate(SABC): Add 100µL of SABC workingsolution to each well. Cover with a new Plate sealer. Incubate for30minutes at 37°C.

5.

Wash: Repeat the aspiration/wash process for five times.

6.

TMB Substrate: Add 90µL of TMB Substrate to each well. Coverwith a new Plate sealer. Incubate for about 10-20 minutes at 37°C.Protect from light. The reaction time can be shortened or extendedaccording to the actual color change, but not more than 30minutes.When apparent gradient appeared in standard wells, you can terminatethe reaction.

7.

Stop: Add 50µL of Stop Solution to each well. Color turn toyellow immediately. The adding order of stop solution should be as thesame as the substrate solution.

8.

OD Measurement: Determine the optical density (OD Value) of each wellat once, using a microplate reader set to 450 nm. You should open themicroplate reader ahead, preheat the instrument, and set the testing parameters.

Sample Type

When carrying out an ELISA assay it is important to prepare your samples in order to achieve the best possible results. Below we have a list of procedures for the preparation of samples for different sample types.

Sample Type Protocol

Serum

If using serum separator tubes, allow samples to clot for 30 minutes at room temperature. Centrifuge for 10 minutes at 1,000x g. Collect the serum fraction and assay promptly or aliquot and store the samples at -80°C. Avoid multiple freeze-thaw cycles.

If serum separator tubes are not being used, allow samples to clot overnight at 2-8°C. Centrifuge for 10 minutes at 1,000x g. Remove serum and assay promptly or aliquot and store the samples at -80°C. Avoid multiple freeze-thaw cycles.

Plasma

Collect plasma using EDTA or heparin as an anticoagulant. Centrifuge samples at 4°C for 15 mins at 1000 × g within 30 mins of collection. Collect the plasma fraction and assay promptly or aliquot and store the samples at -80°C. Avoid multiple freeze-thaw cycles. Note: Over haemolysed samples are not suitable for use with this kit.

Urine & Cerebrospinal Fluid

Collect the urine (mid-stream) in a sterile container, centrifuge for 20 mins at 2000-3000 rpm. Remove supernatant and assay immediately. If any precipitation is detected, repeat the centrifugation step. A similar protocol can be used for cerebrospinal fluid.

Cell culture supernatant

Collect the cell culture media by pipette, followed by centrifugation at 4°C for 20 mins at 1500 rpm. Collect the clear supernatant and assay immediately.

Cell lysates

Solubilize cells in lysis buffer and allow to sit on ice for 30 minutes. Centrifuge tubes at 14,000 x g for 5 minutes to remove insoluble material. Aliquot the supernatant into a new tube and discard the remaining whole cell extract. Quantify total protein concentration using a total protein assay. Assay immediately or aliquot and store at ≤ -20 °C.

Tissue homogenates

The preparation of tissue homogenates will vary depending upon tissue type. Rinse tissue with 1X PBS to remove excess blood & homogenize in 20ml of 1X PBS (including protease inhibitors) and store overnight at ≤ -20°C. Two freeze-thaw cycles are required to break the cell membranes. To further disrupt the cell membranes you can sonicate the samples. Centrifuge homogenates for 5 mins at 5000xg. Remove the supernatant and assay immediately or aliquot and store at -20°C or -80°C.

Tissue lysates

Rinse tissue with PBS, cut into 1-2 mm pieces, and homogenize with a tissue homogenizer in PBS. Add an equal volume of RIPA buffer containing protease inhibitors and lyse tissues at room temperature for 30 minutes with gentle agitation. Centrifuge to remove debris. Quantify total protein concentration using a total protein assay. Assay immediately or aliquot and store at ≤ -20 °C

Breast Milk

Collect milk samples and centrifuge at 10,000 x g for 60 min at 4°C. Aliquot the supernatant and assay. For long term use, store samples at -80°C. Minimize freeze/thaw cycles.

PG Background

PG Background

Progesterone is a vital steroid hormone primarily produced by the ovaries in females, with smaller amounts produced by the adrenal glands and placenta during pregnancy. It plays a crucial role in the female reproductive system and has diverse effects on various physiological processes. Progesterone's levels fluctuate during the menstrual cycle, rising after ovulation to prepare the uterine lining for potential implantation of a fertilized egg. If conception occurs, progesterone continues to be produced to support pregnancy by maintaining the uterine environment. Additionally, progesterone has effects on the nervous system, metabolism, and immune system, contributing to its importance beyond reproduction.

PG Structure and Function

Progesterone is a steroid hormone with a specific chemical structure characterized by a four-ring cyclopentanoperhydrophenanthrene nucleus. Structurally, it is closely related to other steroid hormones, such as testosterone and cortisol. Progesterone's primary function revolves around preparing and maintaining the uterine lining for pregnancy. After ovulation, progesterone is secreted by the corpus luteum in the ovary and helps thicken the endometrial lining, making it suitable for implantation. If fertilization occurs, progesterone continues to be produced, preventing further ovulation and supporting the uterine environment for the developing embryo. If conception does not happen, progesterone levels decrease, leading to the shedding of the uterine lining during menstruation. Progesterone also has a role in suppressing the immune response to prevent rejection of the embryo, and it helps maintain pregnancy by relaxing smooth muscles in the uterus to prevent contractions that could lead to premature labor. Additionally, it affects body temperature regulation and can influence mood and anxiety levels.

PG ELISA KIt FAQs

What is the Human Progesterone ELISA Kit used for?

The Human Progesterone ELISA kit is a laboratory tool employed to quantitatively measure the concentration of progesterone in human serum or plasma samples. Progesterone is a vital hormone in the female reproductive system, influencing processes such as menstrual cycle regulation, pregnancy support, and embryo implantation. Monitoring progesterone levels using this ELISA kit aids in diagnosing and managing conditions related to reproductive health, assessing ovulation, monitoring pregnancy, and investigating hormonal imbalances that can impact fertility and overall hormonal equilibrium.

What are the advantages of using the Human Progesterone ELISA Kit?

The Human Progesterone ELISA Kit offers several advantages, including high sensitivity, accuracy, and reproducibility. It provides a user-friendly and reliable method to quantify Progesterone levels in biological specimens, allowing for precise measurements and robust data analysis.

What sample types are compatible with Human Progesterone ELISA Kit?

The Human Progesterone ELISA Kit is compatible with various sample types, including serum, plasma, cell lysates, and tissue homogenates. It provides flexibility in sample selection, allowing researchers to analyze Progesterone levels in different biological matrices.

What are the storage requirements with Human Progesterone ELISA Kit?

The Human Progesterone ELISA Kit components should be stored according to the instructions provided in the kit manual. Generally, it is recommended to store the kit components at the recommended temperature to ensure their stability and optimal performance.

What should I do if my assay results are not optimal?

If you encounter any issues or have suboptimal assay results, we recommend contacting our dedicated support team for assistance. They will be available to provide troubleshooting guidance, answer your questions, and ensure you achieve the best possible results with the Human Progesterone ELISA Kit.