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Rat Androgen receptor (Ar) ELISA Kit (RTEB0804)

SKU:
RTEB0804
Product Type:
ELISA Kit
Size:
96 Assays
Uniprot:
P15207
ELISA Type:
Sandwich
Reactivity:
Rat
€649
Frequently bought together:

Description

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Rat Androgen receptor (Ar) ELISA Kit

The Rat Androgen Receptor (AR) ELISA Kit is specifically designed for the precise measurement of androgen receptor levels in rat serum, plasma, and cell lysates. This kit offers exceptional sensitivity and specificity, guaranteeing accurate and consistent results for various research purposes.The androgen receptor is a vital transcription factor that plays a crucial role in mediating the effects of androgens in the body, regulating various physiological processes such as development, reproduction, and metabolism.

Dysregulation of the androgen receptor has been linked to a variety of conditions, including prostate cancer and androgen insensitivity syndrome, underscoring its importance as a biomarker for studying these diseases and developing potential treatments.Overall, the Rat Androgen Receptor (AR) ELISA Kit provides a reliable and convenient tool for researchers to assess androgen receptor levels in rat samples, facilitating further understanding of its role in health and disease.

Product Name:Rat Androgen receptor (Ar) ELISA Kit
SKU:RTEB0804
Size:96T
Target:Rat Androgen receptor (Ar)
Synonyms:Dihydrotestosterone receptor, Nuclear receptor subfamily 3 group C member 4, Nr3c4
Assay Type:Sandwich
Detection Method:ELISA
Reactivity:Rat
Detection Range:78-5000pg/mL
Sensitivity:39.9pg/mL
Intra CV:5.3%
Inter CV:7.1%
Linearity:
Sample1:21:41:81:16
Serum(N=5)114-124%106-114%84-93%104-104%
EDTA Plasma(N=5)114-126%103-113%94-104%81-90%
Heparin Plasma(N=5)85-97%94-103%105-115%87-87%
Recovery:
Sample TypeAverage(%)Recovery Range(%)
Serum8781-93
Plasma8983-95
Function:Steroid hormone receptors are ligand-activated transcription factors that regulate eukaryotic gene expression and affect cellular proliferation and differentiation in target tissues. Transcription factor activity is modulated by bound coactivator and corepressor proteins. Transcription activation is down-regulated by NR0B2. Activated, but not phosphorylated, by HIPK3 and ZIPK/DAPK3.
Uniprot:P15207
Sample Type:Serum, plasma, tissue homogenates, cell culture supernates and other biological fluids
Specificity:Natural and recombinant rat Androgen receptor
Sub Unit:Binds DNA as a homodimer. Part of a ternary complex containing AR, EFCAB6/DJBP and PARK7. Interacts with HIPK3 and NR0B2 in the presence of androgen. The ligand binding domain interacts with KAT7/HBO1 in the presence of dihydrotestosterone. Interacts with EFCAB6/DJBP, PELP1, PQBP1, RANBP9, RBAK, SPDEF, SRA1, TGFB1I1, ZNF318 and RREB1. Interacts with ZMIZ1/ZIMP10 and ZMIZ2/ZMIP7 which both enhance its transactivation activity. Interacts with SLC30A9 and RAD54L2/ARIP4. Interacts via the ligand-binding domain with LXXLL and FXXLF motifs from NCOA1, NCOA2, NCOA3, NCOA4 and MAGEA11. The AR N-terminal poly-Gln region binds Ran resulting in enhancement of AR-mediated transactivation. Ran-binding decreases as the poly-Gln length increases. Interacts with HIP1 (via coiled coil domain). Interacts (via ligand-binding domain) with TRIM68. Interacts with TNK2. Interacts with USP26. Interacts with RNF6. Interacts (regulated by RNF6 probably through polyubiquitination) with RNF14; regulates AR transcriptional activity. Interacts with PRMT2 and TRIM24. Interacts with RACK1. Interacts with RANBP10; this interaction enhances dihydrotestosterone-induced AR transcriptional activity. Interacts with PRPF6 in a hormone-independent way; this interaction enhances dihydrotestosterone-induced AR transcriptional activity. Interacts with STK4/MST1. Interacts with ZIPK/DAPK3. Interacts with LPXN. Interacts with MAK. Part of a complex containing AR, MAK and NCOA3. Interacts with CRY1. Interacts with CCAR1 and GATA2.
Subcellular Location:Nucleus Cytoplasm Predominantly cytoplasmic in unligated form but translocates to the nucleus upon ligand-binding. Can also translocate to the nucleus in unligated form in the presence of RACK1.
Storage:Please see kit components below for exact storage details
Note:For research use only
UniProt Protein Function:AR: a nuclear hormone receptor and transcription factor. Regulates gene expression and affects cellular proliferation and differentiation in target tissues. Two splice-variant isoforms have been described.Protein type: DNA-binding; Nuclear receptor; Transcription factorChromosomal Location of Human Ortholog: Xq22Cellular Component: axon; cytoplasm; dendrite; nuclear chromatin; nuclear speck; nucleus; plasma membrane; protein complexMolecular Function: androgen binding; androgen receptor activity; androgen receptor binding; ATPase binding; beta-catenin binding; chromatin binding; DNA binding; DNA binding transcription factor activity; enzyme binding; ligand-dependent nuclear receptor activity; protein binding; protein domain specific binding; receptor binding; ribonucleotide binding; sequence-specific DNA binding; steroid binding; transcription factor binding; zinc ion bindingBiological Process: androgen receptor signaling pathway; animal organ formation; cellular process; copulation; fertilization; in utero embryonic development; intracellular receptor signaling pathway; Leydig cell differentiation; male courtship behavior; male genitalia morphogenesis; male gonad development; male sex differentiation; male somatic sex determination; multicellular organism growth; negative regulation of cell proliferation; negative regulation of epithelial cell proliferation; negative regulation of integrin biosynthetic process; negative regulation of transcription from RNA polymerase II promoter; negative regulation of transcription, DNA-dependent; positive regulation of cell differentiation; positive regulation of cell proliferation; positive regulation of insulin-like growth factor receptor signaling pathway; positive regulation of integrin biosynthetic process; positive regulation of intracellular estrogen receptor signaling pathway; positive regulation of MAPK cascade; positive regulation of NF-kappaB transcription factor activity; positive regulation of phosphorylation; positive regulation of transcription from RNA polymerase II promoter; positive regulation of transcription from RNA polymerase III promoter; positive regulation of transcription, DNA-templated; protein oligomerization; regulation of catalytic activity; regulation of developmental growth; regulation of gene expression; regulation of systemic arterial blood pressure; regulation of transcription from RNA polymerase II promoter; regulation of transcription, DNA-templated; reproductive behavior; reproductive structure development; response to estradiol; response to insulin stimulus; response to testosterone stimulus; single fertilization; skeletal muscle hypertrophy; spermatogenesis; transcription from RNA polymerase II promoter; transcription, DNA-dependent
UniProt Protein Details:
NCBI Summary:binds androgens and activates transcription; required for male sexual development [RGD, Feb 2006]
UniProt Code:P15207
NCBI GenInfo Identifier:6978535
NCBI Gene ID:24208
NCBI Accession:NP_036634.1
UniProt Secondary Accession:P15207,Q63049
UniProt Related Accession:P15207
Molecular Weight:98,218 Da
NCBI Full Name:androgen receptor
NCBI Synonym Full Names:androgen receptor
NCBI Official Symbol:Ar
NCBI Official Synonym Symbols:Tfm; Andr
NCBI Protein Information:androgen receptor
UniProt Protein Name:Androgen receptor
UniProt Synonym Protein Names:Dihydrotestosterone receptor; Nuclear receptor subfamily 3 group C member 4
Protein Family:Allatostatin
UniProt Gene Name:Ar
UniProt Entry Name:
Component Quantity (96 Assays) Storage
ELISA Microplate (Dismountable) 8×12 strips -20°C
Lyophilized Standard 2 -20°C
Sample Diluent 20ml -20°C
Assay Diluent A 10mL -20°C
Assay Diluent B 10mL -20°C
Detection Reagent A 120µL -20°C
Detection Reagent B 120µL -20°C
Wash Buffer 30mL 4°C
Substrate 10mL 4°C
Stop Solution 10mL 4°C
Plate Sealer 5 -

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

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

Allow all reagents to reach room temperature (Please do not dissolve the reagents at 37°C directly). All the reagents should be mixed thoroughly by gently swirling before pipetting. Avoid foaming. Keep appropriate numbers of strips for 1 experiment and remove extra strips from microtiter plate. Removed strips should be resealed and stored at -20°C until the kits expiry date. Prepare all reagents, working standards and samples as directed in the previous sections. Please predict the concentration before assaying. If values for these are not within the range of the standard curve, users must determine the optimal sample dilutions for their experiments. We recommend running all samples in duplicate.

Step
1. Add Sample: Add 100µL of Standard, Blank, or Sample per well. The blank well is added with Sample diluent. Solutions are added to the bottom of micro ELISA plate well, avoid inside wall touching and foaming as possible. Mix it gently. Cover the plate with sealer we provided. Incubate for 120 minutes at 37°C.
2. Remove the liquid from each well, don't wash. Add 100µL of Detection Reagent A working solution to each well. Cover with the Plate sealer. Gently tap the plate to ensure thorough mixing. Incubate for 1 hour at 37°C. Note: if Detection Reagent A appears cloudy warm to room temperature until solution is uniform.
3. Aspirate each well and wash, repeating the process three times. Wash by filling each well with Wash Buffer (approximately 400µL) (a squirt bottle, multi-channel pipette,manifold dispenser or automated washer are needed). Complete removal of liquid at each step is essential. After the last wash, completely remove remaining Wash Buffer by aspirating or decanting. Invert the plate and pat it against thick clean absorbent paper.
4. Add 100µL of Detection Reagent B working solution to each well. Cover with the Plate sealer. Incubate for 60 minutes at 37°C.
5. Repeat the wash process for five times as conducted in step 3.
6. Add 90µL of Substrate Solution to each well. Cover with a new Plate sealer and incubate for 10-20 minutes at 37°C. Protect the plate from light. The reaction time can be shortened or extended according to the actual color change, but this should not exceed more than 30 minutes. When apparent gradient appears in standard wells, user should terminatethe reaction.
7. Add 50µL of Stop Solution to each well. If color change does not appear uniform, gently tap the plate to ensure thorough mixing.
8. Determine the optical density (OD value) of each well at once, using a micro-plate reader set to 450 nm. User should open the micro-plate reader in advance, preheat the instrument, and set the testing parameters.
9. After experiment, store all reagents according to the specified storage temperature respectively until their expiry.

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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