Rat Ferritin heavy chain ELISA Kit

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SKU:
RTFI00065
Product Type:
ELISA Kit
Size:
96 Assays
Uniprot:
P19132
Sensitivity:
3.75ng/ml
Range:
6.25-400ng/ml
ELISA Type:
Sandwich ELISA, Double Antibody
Synonyms:
Fth1, FHC, FTH, PLIF, FTHL6, PIG15, Cell prolifeRation-inducing gene 15 protein, Ferritin H subunit, Ferritin, Heavy Polypeptide
Reactivity:
Rat
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Description

system_update_altデータシートsystem_update_alt安全性データシート
商品名:Rat Fth1 (Ferritin heavy chain) ELISA Kit
製品コード:RTFI00065
サイズ:96 Assays
目標:Rat Fth1
エイリアス:Fth1, FHC, FTH, PLIF, FTHL6, PIG15, Cell prolifeRation-inducing gene 15 protein, Ferritin H subunit, Ferritin, Heavy Polypeptide
反応性:Rat
検出方法:Sandwich ELISA, Double Antibody
感度:3.75ng/ml
範囲:6.25-400ng/ml
保管所:4°C for 6 months
ノート:For Research Use Only
回復:Matrices listed below were spiked with certain level of Rat Fth1 and the recovery rates were calculated by comparing the measured value to the expected amount of Rat Fth1 in samples.
MatrixRecovery range(%)Average(%)
serum(n=5)91-10397
EDTA plasma(n=5)85-10496
UFH plasma(n=5)86-10093
直線性:The linearity of the kit was assayed by testing samples spiked with appropriate concentration of Rat Fth1 and their serial dilutions. The results were demonstrated by the percentage of calculated concentration to the expected.
Sample1:21:41:8
serum(n=5)85-103%93-101%89-104%
EDTA plasma(n=5)83-94%84-101%85-101%
UFH plasma(n=5)82-97%85-97%89-96%
Intra-Assay:CV <8%
Inter-Assay:CV <10%
Uniprot:P19132
UniProt Protein Function:FTL: Stores iron in a soluble, non-toxic, readily available form. Important for iron homeostasis. Iron is taken up in the ferrous form and deposited as ferric hydroxides after oxidation. Also plays a role in delivery of iron to cells. Mediates iron uptake in capsule cells of the developing kidney. Defects in FTL are the cause of hereditary hyperferritinemia-cataract syndrome (HHCS). It is an autosomal dominant disease characterized by early-onset bilateral cataract. Affected patients have elevated level of circulating ferritin. HHCS is caused by mutations in the iron responsive element (IRE) of the FTL gene. Defects in FTL are the cause of neurodegeneration with brain iron accumulation type 3 (NBIA3); also known as adult-onset basal ganglia disease. It is a movement disorder with heterogeneous presentations starting in the fourth to sixth decade. It is characterized by a variety of neurological signs including parkinsonism, ataxia, corticospinal signs, mild nonprogressive cognitive deficit and episodic psychosis. It is linked with decreased serum ferritin levels. Belongs to the ferritin family.
UniProt Protein Details:

Protein type:Oxidoreductase

Chromosomal Location of Human Ortholog: 19q13.33

Cellular Component: ferritin complex; membrane; cytosol

Molecular Function:identical protein binding; protein binding; ferric iron binding; iron ion binding

Biological Process: receptor-mediated endocytosis; iron ion homeostasis; cellular iron ion homeostasis; post-Golgi vesicle-mediated transport; transmembrane transport; iron ion transport

Disease: Hyperferritinemia With Or Without Cataract; L-ferritin Deficiency; Neurodegeneration With Brain Iron Accumulation 3

NCBI Summary:This gene encodes the light subunit of the ferritin protein. Ferritin is the major intracellular iron storage protein in prokaryotes and eukaryotes. It is composed of 24 subunits of the heavy and light ferritin chains. Variation in ferritin subunit composition may affect the rates of iron uptake and release in different tissues. A major function of ferritin is the storage of iron in a soluble and nontoxic state. Defects in this light chain ferritin gene are associated with several neurodegenerative diseases and hyperferritinemia-cataract syndrome. This gene has multiple pseudogenes. [provided by RefSeq, Jul 2008]
UniProt Code:P19132
NCBI GenInfo Identifier:20149498
NCBI Gene ID:2512
NCBI Accession:NP_000137.2
UniProt Secondary Accession:P19132,P02794, P02793, P19132,
UniProt Related Accession:P02792
Molecular Weight:
NCBI Full Name:ferritin light chain
NCBI Synonym Full Names:ferritin light chain
NCBI Official Symbol:FTL  
NCBI Official Synonym Symbols:LFTD; NBIA3  
NCBI Protein Information:ferritin light chain
UniProt Protein Name:Ferritin light chain
Protein Family:Ferritin
UniProt Gene Name:FTL  
UniProt Entry Name:FRIL_HUMAN
ステッププロトコル
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.

ELISAアッセイを実施する場合、可能な限り最良の結果を達成するためにサンプルを準備することが重要です。以下に、さまざまなサンプルタイプのサンプルを準備するための手順のリストを示します。

サンプルタイプ プロトコル
血清

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.

プラズマ 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.
尿および脳脊髄液 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.
細胞培養上清 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.
細胞溶解物 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.
組織ホモジネート 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.
組織溶解物 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.
母乳 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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