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Human Cavin 1(Polymerase I and transcript release factor) ELISA Kit (HUFI03151)

SKU:
HUFI03151
Product Type:
ELISA Kit
Size:
96 Assays
Uniprot:
Q6NZI2
Sensitivity:
46.875pg/ml
Range:
78.125-5000pg/ml
ELISA Type:
Sandwich
Synonyms:
Polymerase I and transcript release factor
Reactivity:
Human
€599
Frequently bought together:

Description

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Human Cavin 1 (Polymerase I and transcript release factor) ELISA Kit

The Human CAVIN-1 (Polymerase I and Transcript Release Factor) ELISA Kit is a powerful tool for researchers looking to accurately measure CAVIN-1 levels in human samples such as serum, plasma, and cell culture supernatants. With its high sensitivity and specificity, this kit delivers reliable and reproducible results, making it an excellent choice for a variety of research applications.CAVIN-1 is a key protein involved in various cellular processes, including polymerase I and transcript release factor functions.

Its role in cell signaling and membrane trafficking makes it a pivotal player in multiple pathways, making it a valuable biomarker for studying diseases like cancer, cardiovascular disorders, and metabolic syndromes.By utilizing the Human CAVIN-1 ELISA Kit, researchers can gain valuable insights into the role of CAVIN-1 in disease progression and potential therapeutic interventions, advancing our understanding of complex biological processes.

Product Name:Human Cavin 1(Polymerase I and transcript release factor) ELISA Kit
Product Code:HUFI03151
Size:96 Assays
Alias:Polymerase I and transcript release factor
Detection method:Sandwich ELISA, Double Antibody
Application:This immunoassay kit allows for the in vitro quantitative determination of Human Cavin 1 concentrations in serum plasma and other biological fluids.
Sensitivity:46.875pg/ml
Range:78.125-5000pg/ml
Storage:4°C for 6 months
Note:For Research Use Only
Recovery:Matrices listed below were spiked with certain level of Human Cavin 1 and the recovery rates were calculated by comparing the measured value to the expected amount of Human Cavin 1 in samples. Enquire for more information.
Linearity:The linearity of the kit was assayed by testing samples spiked with appropriate concentration of Human Cavin 1 and their serial dilutions. The results were demonstrated by the percentage of calculated concentration to the expected. Enquire for more information.
CV(%):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
UniprotQ6NZI2
UniProt Protein Function:PTRF: Plays an important role in caveolae formation and organization. Required for the sequestration of mobile caveolin into immobile caveolae. Termination of transcription by RNA polymerase I involves pausing of transcription by TTF1, and the dissociation of the transcription complex, releasing pre-rRNA and RNA polymerase I from the template. PTRF is required for dissociation of the ternary transcription complex. Defects in PTRF are the cause of congenital generalized lipodystrophy type 4 (CGL4). It is a disorder characterized by the association of congenital generalized lipodystrophy with muscular dystrophy and cardiac anomalies. Congenital generalized lipodystrophy is characterized by a near complete absence of adipose tissue, extreme insulin resistance, hypertriglyceridemia, hepatic steatosis and early onset of diabetes. Belongs to the PTRF/SDPR family. 3 isoforms of the human protein are produced by alternative splicing.
UniProt Protein Details:Protein type:Transcription initiation complex

Chromosomal Location of Human Ortholog: 17q21.2

Cellular Component: nucleoplasm; protein complex; mitochondrion; intracellular membrane-bound organelle; endoplasmic reticulum; cytoplasm; plasma membrane; caveola; cytosol; nucleus

Molecular Function:protein binding; rRNA primary transcript binding

Biological Process: regulation of transcription, DNA-dependent; transcription from RNA polymerase I promoter; gene expression; termination of RNA polymerase I transcription; transcription initiation from RNA polymerase I promoter

Disease: Lipodystrophy, Congenital Generalized, Type 4

NCBI Summary:This gene encodes a protein that enables the dissociation of paused ternary polymerase I transcription complexes from the 3' end of pre-rRNA transcripts. This protein regulates rRNA transcription by promoting the dissociation of transcription complexes and the reinitiation of polymerase I on nascent rRNA transcripts. This protein also localizes to caveolae at the plasma membrane and is thought to play a critical role in the formation of caveolae and the stabilization of caveolins. This protein translocates from caveolae to the cytoplasm after insulin stimulation. Caveolae contain truncated forms of this protein and may be the site of phosphorylation-dependent proteolysis. This protein is also thought to modify lipid metabolism and insulin-regulated gene expression. Mutations in this gene result in a disorder characterized by generalized lipodystrophy and muscular dystrophy. [provided by RefSeq, Nov 2009]
UniProt Code:Q6NZI2
NCBI GenInfo Identifier:56749614
NCBI Gene ID:284119
NCBI Accession:Q6NZI2.1
UniProt Secondary Accession:Q6NZI2,O00535, Q6GMY1, Q96H74, Q9BT85, Q9HAP4, B2RAW7
UniProt Related Accession:Q6NZI2
Molecular Weight:390
NCBI Full Name:Polymerase I and transcript release factor
NCBI Synonym Full Names:polymerase I and transcript release factor
NCBI Official Symbol:PTRF  
NCBI Official Synonym Symbols:CGL4; CAVIN; CAVIN1; FKSG13; cavin-1  
NCBI Protein Information:polymerase I and transcript release factor; PTRF; TTF-I interacting peptide 12; RNA polymerase I and transcript release factor
UniProt Protein Name:Polymerase I and transcript release factor
UniProt Synonym Protein Names:Cavin-1
Protein Family:Polymerase I and transcript release factor
UniProt Gene Name:PTRF  
UniProt Entry Name:PTRF_HUMAN

*Note: Protocols are specific to each batch/lot. For the correct 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.

StepProtocol
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 TypeProtocol
SerumIf 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.

PlasmaCollect 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 FluidCollect 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 supernatantCollect 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 lysatesSolubilize 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 homogenatesThe 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 lysatesRinse 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 MilkCollect 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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