Mouse GR beta / Glucocorticoid Receptor Beta ELISA Kit
- SKU:
- MOFI00869
- Product Type:
- ELISA Kit
- Size:
- 96 Assays
- Uniprot:
- P06537-5
- Sensitivity:
- 9.375pg/ml
- Range:
- 15.625-1000pg/ml
- ELISA Type:
- Sandwich
- Synonyms:
- GRBeta
- Reactivity:
- Mouse
Description
Mouse GR beta / Glucocorticoid Receptor Beta ELISA Kit
GRβ is a splice variant of the glucocorticoid receptor gene that has been shown to have intrinsic activities and modulate gene expression and physiological functions in a cell-specific manner. The Assay Genie Mouse GR beta ELISA kit enables accurate measurement of GRβ levels, providing valuable insights into its role in cellular and physiological processes.
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: |
Mouse GR beta / Glucocorticoid Receptor Beta ELISA Kit |
|
Product Code: |
MOFI00869 |
|
Size: |
96 Assays |
|
Alias: |
GRbeta |
|
Detection Method: |
Sandwich ELISA |
|
Application: |
This immunoassay kit allows for the in vitro quantitative determination of Mouse GRbeta concentrations in serum plasma and other biological fluids. |
|
Sensitivity: |
9.375pg/ml |
|
Range: |
15.625-1000pg/ml |
|
Storage: |
4°C for 6 months |
|
Note: |
For Research Use Only |
Additional Information
|
Recovery |
Matrices listed below were spiked with certain level of Mouse GRbeta and the recovery rates were calculated by comparing the measured value to the expected amount of Mouse GRbeta in samples.
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Linearity: |
The linearity of the kit was assayed by testing samples spiked with appropriate concentration of Mouse GRbeta and their serial dilutions. The results were demonstrated by the percentage of calculated concentration to the expected.
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CV(%) |
Intra Assay <8 Inter Assay <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 exact instructions please follow the protocol included in your kit.
Before adding to wells, equilibrate the SABC working solution and TMB substrate for at least 30 min at 37°C. When diluting samples and reagents, they must be mixed completely and evenly. 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-coated plate respectively, and then, record their positions. It is recommended to measure each standard and sample in duplicate. Wash plate 2 times before adding standard, sample and control (zero) wells! |
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2. |
Aliquot 0.1ml standard solutions into the standard wells. |
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3. |
Add 0.1 ml of Sample / Standard dilution buffer into the control (zero) well. |
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4. |
Add 0.1 ml of properly diluted sample ( Human serum, plasma, tissue homogenates and other biological fluids.) into test sample wells. |
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5. |
Seal the plate with a cover and incubate at 37 °C for 90 min. |
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6. |
Remove the cover and discard the plate content, clap the plate on the absorbent filter papers or other absorbent material. Do NOT let the wells completely dry at any time. Wash plate X2. |
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7. |
Add 0.1 ml of Biotin- detection antibody working solution into the above wells (standard, test sample & zero wells). Add the solution at the bottom of each well without touching the side wall. |
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8. |
Seal the plate with a cover and incubate at 37°C for 60 min. |
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9. |
Remove the cover, and wash plate 3 times with Wash buffer. Let wash buffer rest in wells for 1 min between each wash. |
|
10. |
Add 0.1 ml of SABC working solution into each well, cover the plate and incubate at 37°C for 30 min. |
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11. |
Remove the cover and wash plate 5 times with Wash buffer, and each time let the wash buffer stay in the wells for 1-2 min. |
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12. |
Add 90 µl of TMB substrate into each well, cover the plate and incubate at 37°C in dark within 10-20 min. (Note: This incubation time is for reference use only, the optimal time should be determined by end user.) And the shades of blue can be seen in the first 3-4 wells (with most concentrated standard solutions), the other wells show no obvious color. |
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13. |
Add 50 µl of Stop solution into each well and mix thoroughly. The color changes into yellow immediately. |
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14. |
Read the O.D. absorbance at 450 nm in a microplate reader immediately after adding the stop solution. |
Sample Preparation
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. |
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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. |
Glucocorticoid Receptor Beta Background
Glucocorticoids and their receptors
Glucocorticoids (GCs) are hormones produced by the adrenal cortex in response to activation of the hypothalamic-pituitary-adrenal axis. They play a crucial role in regulating various physiological processes and exhibit anti-inflammatory and immunosuppressive effects. Synthetic GCs have been widely used in the treatment of inflammatory conditions and organ transplant rejection prevention.
GCs exert their effects through the glucocorticoid receptor (GR), which is expressed throughout the body and mediates diverse cellular responses. The GR-mediated responses can vary in strength and specificity across different cell types and tissues. This variability is further expanded by the existence of multiple GR isoforms resulting from alternative translation initiation or alternative splicing.
Glucocorticoid Receptor isoforms
One significant splicing event near the end of the primary GR transcript generates two major isoforms: GRα and GRβ. GRα is the predominant isoform and is responsible for mediating the actions of GCs. On the other hand, GRβ has been implicated in interfering with GRα-mediated activities. While GRα is the classic receptor involved in GC signaling, GRβ's role is associated with the impairment of GRα-mediated functions.
Glucocorticoid receptor beta
The glucocorticoid receptor beta (GRβ) is a splice variant of the glucocorticoid receptor (GR) gene. It is generated by alternative splicing of the primary transcript of the GR gene, resulting in the production of a truncated protein isoform. The GRβ lacks the C-terminal part of the GR, including the ligand-binding domain, which is critical for hormone binding.
Glucocorticoid Function and Clinical Significance
Initially, GRβ was considered a dominant-negative regulator of the glucocorticoid signaling pathway. It competes with the full-length glucocorticoid receptor (GRα) for binding to glucocorticoid response elements (GREs) on target genes. This competition could lead to the suppression of GRα-mediated gene transcription, resulting in a reduced glucocorticoid response.
Recent evidence has challenged the previously held beliefs regarding glucocorticoid receptor beta (GRβ) and has demonstrated that these concepts are cell-specific. The new findings indicate that GRβ is capable of binding ligands like RU486. Additionally, GRβ has intrinsic activities that allow it to modulate gene expression and influence various physiological functions.
Glucocorticoid receptor beta (GRβ) has been implicated in various diseases and cellular functions. Inflammatory disorders frequently exhibit GRβ expression, which has been associated with glucocorticoid (GC) insensitivity. However, recent studies have shown that GRβ can directly regulate gene expression independently of its antagonism of GRα, exhibiting intrinsic activities. GRβ has been linked to metabolic functions such as gluconeogenesis and lipid storage, as well as inflammation modulation through the activation of pro-inflammatory pathways. It also plays a role in cell migration and proliferation, influencing the expression of genes associated with extracellular matrix interactions and the Wnt/β-catenin and PTEN/PI3K/Akt signaling pathways. Additionally, GRβ may affect apoptosis in a cell-specific manner. Targeting GRβ may hold promise for the treatment of diseases where it plays a pathogenic role, providing new insights into cellular and physiological functions.
Mouse GR Beta ELISA Kits FAQs
Q: What is the purpose of the Mouse GR beta ELISA Kit?
The Mouse GR beta ELISA Kit is specifically designed for quantitative detection of glucocorticoid receptor beta (GRβ) protein levels in mouse samples. GRβ is a splice variant of the glucocorticoid receptor gene that has been shown to have intrinsic activities and modulate gene expression and physiological functions in a cell-specific manner. This ELISA kit enables accurate measurement of GRβ levels, providing valuable insights into its role in cellular and physiological processes.
Q: What type of samples can be used with the mouse GR beta ELISA Kit?
The mouse GR beta 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 study GRβ protein levels in different biological materials.
Q: Is the kit specific to mouse samples only?
es, the mouse GR beta ELISA Kit is specifically designed and validated for the detection of GRβ protein levels in mouse samples. It is important to note that the kit is not suitable for use with samples from other species without proper validation.
Q: Where can I find additional technical support or assistance with the Mouse GR beta ELISA kit?
For any technical inquiries or assistance regarding the Mouse GR beta ELISA kit, you can reach out to our team. They will be available to answer your questions and provide the necessary guidance to ensure a successful experiment.
Related Products
| Mouse GR alpha / Glucocorticoid Receptor Alpha ELISA Kit | |
|---|---|
| ELISA TYPE: | Sandwich ELISA, Double Antibody |
| SENSITIVITY: | 9.375pg/ml |
| RANGE: | 15.625-1000pg/ml |
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Human GR beta / Glucocorticoid Receptor Beta ELISA Kit
Mouse GR beta (glucocorticoid receptor beta) CLIA Kit (MOES00292)
