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Rat NFIL-3 ELISA Kit (RTFI00987)

SKU:
RTFI00987
Product Type:
ELISA Kit
Size:
96 Assays
Uniprot:
Q6IMZ0
Sensitivity:
0.094ng/ml
Range:
0.156-10ng/ml
ELISA Type:
Sandwich
Synonyms:
NFIL3, E4BP4, IL3BP1, NF-IL3A, E4BP4IL3BP1, Interleukin-3 promoter transcriptional activator, Interleukin-3-binding protein 1, NF-IL3A, NFIL3AE4 promoter-binding protein 4, nuclear factor interleukin 3 regulated protein, Transcriptional activator NF-
Reactivity:
Rat
Research Area:
Cell Biology
€649
Frequently bought together:

Description

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Rat NFIL-3 ELISA Kit

The Rat NFIL3 ELISA Kit is specifically designed to accurately measure NFIL3 levels in rat serum, plasma, and cell culture supernatants. With its high sensitivity and specificity, this kit provides researchers with reliable and reproducible results, making it an ideal tool for various research applications.NFIL3, also known as E4BP4, is a key transcription factor involved in regulating immune responses and circadian rhythms. It plays a crucial role in maintaining immune homeostasis and has been implicated in various diseases, including autoimmune disorders and cancer.

By measuring NFIL3 levels, researchers can gain valuable insights into the role of this protein in disease pathogenesis and potential therapeutic interventions.Overall, the Rat NFIL3 ELISA Kit offers a valuable tool for studying NFIL3 biology and its implications in disease research, providing researchers with a reliable means of quantifying NFIL3 levels in biological samples.

Product Name:Rat NFIL3 (Nuclear Factor, Interleukin 3 Regulated) ELISA Kit
Product Code:RTFI00987
Size:96 Assays
Target:Rat NFIL3
Alias:NFIL3, E4BP4, IL3BP1, NF-IL3A, E4BP4IL3BP1, Interleukin-3 promoter transcriptional activator, Interleukin-3-binding protein 1, NF-IL3A, NFIL3AE4 promoter-binding protein 4, nuclear factor interleukin 3 regulated protein, Transcriptional activator NF-IL3A
Reactivity:Rat
Detection Method:Sandwich ELISA, Double Antibody
Sensitivity:0.094ng/ml
Range:0.156-10ng/ml
Storage:4°C for 6 months
Note:For Research Use Only
Recovery:Matrices listed below were spiked with certain level of Rat NFIL3 and the recovery rates were calculated by comparing the measured value to the expected amount of Rat NFIL3 in samples.
MatrixRecovery range(%)Average(%)
serum(n=5)90-10296
EDTA plasma(n=5)90-10195
UFH plasma(n=5)86-10294
Linearity:The linearity of the kit was assayed by testing samples spiked with appropriate concentration of Rat NFIL3 and their serial dilutions. The results were demonstrated by the percentage of calculated concentration to the expected.
Sample1:21:41:8
serum(n=5)86-99%87-102%87-98%
EDTA plasma(n=5)84-101%84-101%83-98%
UFH plasma(n=5)92-98%82-97%83-100%
Intra-Assay:CV <8%
Inter-Assay:CV <10%
Uniprot:Q6IMZ0
UniProt Protein Function:E4BP4: Acts as a transcriptional regulator that recognizes and binds to the sequence 5'-[GA]TTA[CT]GTAA[CT]-3', a sequence present in many cellular and viral promoters. Represses transcription from promoters with activating transcription factor (ATF) sites. Represses promoter activity in osteoblasts. Represses transcriptional activity of PER1. Represses transcriptional activity of PER2 via the B- site on the promoter. Activates transcription from the interleukin-3 promoter in T-cells. Competes for the same consensus-binding site with PAR DNA-binding factors (DBP, HLF and TEF). Component of the circadian clock that acts as a negative regulator for the circadian expression of PER2 oscillation in the cell-autonomous core clock. Protects pro-B cells from programmed cell death. Belongs to the bZIP family. NFIL3 subfamily.
UniProt Protein Details:

Protein type:DNA-binding; Transcription factor

Cellular Component: nucleus

Molecular Function:transcription factor activity

Biological Process: transcription from RNA polymerase II promoter; circadian rhythm; immune response; negative regulation of transcription from RNA polymerase II promoter

NCBI Summary:The protein encoded by this gene is a transcriptional regulator that binds as a homodimer to activating transcription factor (ATF) sites in many cellular and viral promoters. The encoded protein represses Per1 and Per2 expression and therefore plays a role in the regulation of circadian rhythm. [provided by RefSeq, Feb 2014]
UniProt Code:Q6IMZ0
NCBI GenInfo Identifier:48675850
NCBI Gene ID:114519
NCBI Accession:NP_446179.2
UniProt Secondary Accession:Q6IMZ0,Q923M2,
UniProt Related Accession:Q6IMZ0
Molecular Weight:50,781 Da
NCBI Full Name:nuclear factor interleukin-3-regulated protein
NCBI Synonym Full Names:nuclear factor, interleukin 3 regulated
NCBI Official Symbol:Nfil3  
NCBI Official Synonym Symbols:E4BP4  
NCBI Protein Information:nuclear factor interleukin-3-regulated protein
UniProt Protein Name:Nuclear factor interleukin-3-regulated protein
UniProt Synonym Protein Names:E4 promoter-binding protein 4
Protein Family:Nuclear factor interleukin-3-regulated protein
UniProt Gene Name:Nfil3  
UniProt Entry Name:NFIL3_RAT
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 clotovernight at 2-8°C. Centrifuge for 10 minutes at 1,000x g. Removeserum 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 anti-coagulant. 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 & homogenizein 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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