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Mouse ER alpha / NR3A1 / Estrogen Receptor alpha ELISA Kit

SKU:
MOFI00794
Product Type:
ELISA Kit
Size:
96 Assays
Uniprot:
P19785
Sensitivity:
0.188ng/ml
Range:
0.313-20ng/ml
ELISA Type:
Sandwich
Synonyms:
ERalpha, ESR1, NR3A1, ER, ER alpha, Era, ER-alpha, ESRESRA, Estradiol receptor, estrogen receptor, estrogen receptor 1, estrogen receptor alpha delta 3*4, 56, 7*, 819-2 isoform, estrogen receptor alpha delta 4 +49 isoform, estrogen receptor alpha del
Reactivity:
Mouse
€649
Frequently bought together:

Description

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Mouse ER alpha/NR3A1/Estrogen Receptor alpha ELISA Kit

The Mouse ER Alpha (NR3A1) Estrogen Receptor Alpha ELISA Kit is specifically designed for the accurate detection of estrogen receptor alpha levels in mouse serum, plasma, and cell culture supernatants. This kit offers high sensitivity and specificity, ensuring precise and reliable results for a variety of research applications.Estrogen receptor alpha is a key protein involved in regulating the effects of estrogen in the body, influencing various physiological processes such as growth, reproduction, and metabolism.

Dysregulation of estrogen receptor alpha has been linked to a number of conditions including breast cancer, osteoporosis, and cardiovascular diseases, making it an important biomarker for studying these diseases and developing targeted therapies.With its advanced technology and easy-to-use format, the Mouse ER Alpha (NR3A1) Estrogen Receptor Alpha ELISA Kit is an essential tool for researchers looking to explore the role of estrogen receptor alpha in health and disease in mouse models.

Product Name:Mouse ER alpha / NR3A1 / Estrogen Receptor alpha ELISA Kit
Product Code:MOFI00794
Size:96 Assays
Alias:ERalpha, ESR1, NR3A1, ER, ER alpha, Era, ER-alpha, ESRESRA, Estradiol receptor, estrogen receptor, estrogen receptor 1, estrogen receptor alpha delta 3*4, 56, 7*, 819-2 isoform, estrogen receptor alpha delta 4 +49 isoform, estrogen receptor alpha delta 4*5, 6, 7*, 654 isoform, NR3A1DKFZp686N23123, Nuclear receptor subfamily 3 group A member 1
Detection Method:Sandwich ELISA
Application:This immunoassay kit allows for the in vitro quantitative determination of Mouse ERalpha 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
Recovery: Matrices listed below were spiked with certain level of Mouse ERalpha and the recovery rates were calculated by comparing the measured value to the expected amount of Mouse ERalpha in samples.
MatrixRecovery range(%)Average(%)
serum(n=5)87-10496
EDTA plasma(n=5)87-9591
UFH plasma(n=5)85-10093
Linearity:The linearity of the kit was assayed by testing samples spiked with appropriate concentration of Mouse ERalpha and their serial dilutions. The results were demonstrated by the percentage of calculated concentration to the expected.
Sample1:21:41:8
serum(n=5)92-100%86-102%86-99%
EDTA plasma(n=5)85-100%85-99%83-99%
UFH plasma(n=5)84-98%85-100%80-99%
Intra Assay:CV <8%
Inter Assay:CV <10%
ComponentQuantityStorage
ELISA Microplate (Dismountable)8×12 strips4°C for 6 months
Lyophilized Standard24°C/-20°C
Sample/Standard Dilution Buffer20ml4°C
Biotin-labeled Antibody(Concentrated)120ul4°C (Protect from light)
Antibody Dilution Buffer10ml4°C
HRP-Streptavidin Conjugate(SABC)120ul4°C (Protect from light)
SABC Dilution Buffer10ml4°C
TMB Substrate10ml4°C (Protect from light)
Stop Solution10ml4°C
Wash Buffer(25X)30ml4°C
Plate Sealer5-

Other materials and equipment 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
UniprotP19785
UniProt Protein Function:Nuclear hormone receptor. The steroid hormones and their receptors are involved in the regulation of eukaryotic gene expression and affect cellular proliferation and differentiation in target tissues. Ligand-dependent nuclear transactivation involves either direct homodimer binding to a palindromic estrogen response element (ERE) sequence or association with other DNA-binding transcription factors, such as AP-1/c-Jun, c-Fos, ATF-2, Sp1 and Sp3, to mediate ERE-independent signaling. Ligand binding induces a conformational change allowing subsequent or combinatorial association with multiprotein coactivator complexes through LXXLL motifs of their respective components. Mutual transrepression occurs between the estrogen receptor (ER) and NF-kappa-B in a cell-type specific manner. Decreases NF-kappa-B DNA-binding activity and inhibits NF-kappa-B-mediated transcription from the IL6 promoter and displace RELA/p65 and associated coregulators from the promoter. Recruited to the NF-kappa-B response element of the CCL2 and IL8 promoters and can displace CREBBP. Present with NF-kappa-B components RELA/p65 and NFKB1/p50 on ERE sequences. Can also act synergistically with NF-kappa-B to activate transcription involving respective recruitment adjacent response elements; the function involves CREBBP. Can activate the transcriptional activity of TFF1. Also mediates membrane-initiated estrogen signaling involving various kinase cascades. Essential for MTA1-mediated transcriptional regulation of BRCA1 and BCAS3.
UniProt Protein Details:
NCBI Summary:This gene encodes an estrogen receptor, a member of the nuclear hormone family of intracellular receptors. The encoded protein, activated by the sex hormone estrogen, is a transcription factor composed of several domains important for hormone binding, DNA binding, and activation of transcription. Estrogen and its receptors are essential for sexual development and reproductive function, but also play a role in other tissues such as bone. Similar genes in human have been implicated in pathological processes including breast cancer, endometrial cancer, and osteoporosis. Alternative splicing results in multiple transcript variants. [provided by RefSeq, Oct 2014]
UniProt Code:P19785
NCBI GenInfo Identifier:700274121
NCBI Gene ID:13982
NCBI Accession:NP_001289460.1
UniProt Secondary Accession:P19785,Q9JJT5, Q9QY51, Q9QY52
UniProt Related Accession:P19785
Molecular Weight:66,955 Da
NCBI Full Name:estrogen receptor isoform 1
NCBI Synonym Full Names:estrogen receptor 1 (alpha)
NCBI Official Symbol:Esr1  
NCBI Official Synonym Symbols:ER; ERa; ESR; Estr; Estra; Nr3a1; ERalpha; ER-alpha  
NCBI Protein Information:estrogen receptor
UniProt Protein Name:Estrogen receptor
UniProt Synonym Protein Names:ER-alpha; Estradiol receptor; Nuclear receptor subfamily 3 group A member 1
Protein Family:GTPase
UniProt Gene Name:Esr1  
UniProt Entry Name:

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

StepProcedure
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!
2.Aliquot 0.1ml standard solutions into the standard wells.
3. Add 0.1 ml of Sample / Standard dilution buffer into the control (zero) well.
4. Add 0.1 ml of properly diluted sample (Human serum, plasma, tissue homogenates and other biological fluids.) into test sample wells.
5.Seal the plate with a cover and incubate at 37 °C for 90 min.
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.
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.
8.Seal the plate with a cover and incubate at 37°C for 60 min.
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.
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.
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.
13. Add 50 µL of Stop solution into each well and mix thoroughly. The color changes into yellow immediately.
14. Read the O.D. absorbance at 450 nm in a microplate reader immediately after adding the stop solution.

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.

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