null

Mouse Retrotransposon-derived protein PEG10 (Peg10) ELISA Kit (MOEB2144)

SKU:
MOEB2144
Product Type:
ELISA Kit
Size:
96 Assays
Uniprot:
Q7TN75
ELISA Type:
Sandwich
Reactivity:
Mouse
€649
Frequently bought together:

Description

system_update_altDatasheetsystem_update_altMSDS

Mouse Retrotransposon-derived protein PEG10 (Peg10) ELISA Kit

The Mouse PEG10 ELISA Kit is specifically designed for the precise measurement of PEG10 levels in mouse serum, plasma, and cell culture supernatants. With its high sensitivity and specificity, this kit provides accurate and reproducible results, making it suitable for various research applications.PEG10, a protein derived from mouse retrotransposon, plays a crucial role in various cellular processes and has been implicated in cancer development and progression.

By detecting and quantifying PEG10 levels, researchers can better understand the molecular mechanisms underlying tumorigenesis and potentially identify novel therapeutic targets for cancer treatment.Overall, the Mouse PEG10 ELISA Kit offers a reliable tool for investigating the role of PEG10 in cancer biology and other related fields, providing valuable insights for further research and potential clinical applications.

Product Name:Mouse Retrotransposon-derived protein PEG10 (Peg10) ELISA Kit
SKU:MOEB2144
Size:96T
Target:Mouse Retrotransposon-derived protein PEG10 (Peg10)
Synonyms:Embryonal carcinoma differentiation regulated protein, Mammalian retrotransposon-derived protein 2, Myelin expression factor 3, Myelin expression factor 3-like protein 1, Paternally expressed gene 10 protein, Retrotransposon gag domain-containing protein 3, Retrotransposon-derived gag-like polyprotein, Ty3/Gypsy-like protein, MyEF-3, MEF3-like protein 1, Edr, Mar2, Mart2, Mef3l1, Rgag3
Assay Type:Sandwich
Detection Method:ELISA
Reactivity:Mouse
Detection Range:0.156-10ng/mL
Sensitivity:0.072ng/mL
Intra CV:5.3%
Inter CV:7.6%
Linearity:
Sample1:21:41:81:16
Serum(N=5)96-106%98-109%108-118%104-114%
EDTA Plasma(N=5)103-115%109-117%88-98%110-120%
Heparin Plasma(N=5)97-107%106-106%100-110%97-107%
Recovery:
Sample TypeAverage(%)Recovery Range(%)
Serum8781-93
Plasma8983-95
Function:May have a role in cell growth promotion, apoptotic resistance and hepatoma formation. Inhibits the TGF-beta signaling by interacting with the TGF-beta receptor ALK1. When overexpressed, induces the formation of cellular extension, such as filipodia in association with ALK1 (By similarity). Involved at the immediate early stage of adipocyte differentiation. May bind to the 5'-GCCTGTCTTT-3' DNA sequence of the MB1 domain in the myelin basic protein (MBP) promoter.
Uniprot:Q7TN75
Sample Type:Serum, plasma, tissue homogenates, cell culture supernates and other biological fluids
Specificity:Natural and recombinant mouse Retrotransposon-derived protein PEG10
Sub Unit:Interacts with ALK1, SIAH1 and SIAH2.
Research Area:Epigenetics
Subcellular Location:Nucleus Cytoplasm Detected predominantly in the cytoplasm of breast and prostate carcinomas, in hepatocellular carcinoma (HCC) and B-cell chronic lymphocytic leukemia (B-CLL) cells. Colocalized with ALK1 (By similarity).
Storage:Please see kit components below for exact storage details
Note:For research use only
UniProt Protein Function:PEG10: Prevents apoptosis in hepatocellular carcinoma (HCC) cells through interaction with SIAH1, a mediator of apoptosis. May also have a role in cell growth promotion and hepatoma formation. Inhibits the TGF-beta signaling by interacting with the TGF-beta receptor ALK1. When overexpressed, induces the formation of cellular extension, such as filipodia in association with ALK1. Involved at the immediate early stage of adipocyte differentiation. May bind to the 5'-GCCTGTCTTT-3' DNA sequence of the MB1 domain in the myelin basic protein (MBP) promoter. Up-regulated by MYC and dihydrotestosteron (DHT). Interacts with ALK1, SIAH1 and SIAH2. 2 isoforms of the human protein are produced by ribosomal frameshifting.Protein type: RNA-binding; Apoptosis; Cell cycle regulation; Cell development/differentiationCellular Component: cytoplasm; nucleusMolecular Function: nucleic acid binding; DNA binding; zinc ion binding; metal ion bindingBiological Process: apoptosis; cell differentiation; negative regulation of transforming growth factor beta receptor signaling pathway; placenta development
UniProt Protein Details:
NCBI Summary:This is a paternally expressed imprinted gene that is thought to have been derived from the Ty3/Gypsy family of retrotransposons. It contains two overlapping open reading frames, RF1 and RF2, and expresses two proteins: a shorter, gag-like protein (with a CCHC-type zinc finger domain) from RF1; and a longer, gag/pol-like fusion protein (with an additional aspartic protease motif) from RF1/RF2 by -1 translational frameshifting (-1 FS). While -1 FS has been observed in RNA viruses and transposons in both prokaryotes and eukaryotes, this gene represents the first example of -1 FS in a eukaryotic cellular gene. This gene is highly conserved across mammalian species and retains the heptanucleotide (GGGAAAC) and pseudoknot elements required for -1 FS. It is expressed in adult and embryonic tissues (most notably in placenta) and reported to have a role in cell proliferation, differentiation, apoptosis and cancer development. Knockout mice lacking this gene showed early embryonic lethality with placental defects, indicating the importance of this gene in embryonic development. [provided by RefSeq, Oct 2014]
UniProt Code:Q7TN75
NCBI GenInfo Identifier:98985812
NCBI Gene ID:170676
NCBI Accession:NP_001035701.1
UniProt Secondary Accession:Q7TN75,Q9EQ11
UniProt Related Accession:Q7TN75
Molecular Weight:42,328 Da
NCBI Full Name:retrotransposon-derived protein PEG10 isoform 2
NCBI Synonym Full Names:paternally expressed 10
NCBI Official Symbol:Peg10
NCBI Official Synonym Symbols:Edr; HB-1; Mar2; MEF3L; Mart2; MyEF-3; AA407948
NCBI Protein Information:retrotransposon-derived protein PEG10
UniProt Protein Name:Retrotransposon-derived protein PEG10
UniProt Synonym Protein Names:Embryonal carcinoma differentiation regulated protein; Mammalian retrotransposon-derived protein 2; Myelin expression factor 3; MyEF-3; Myelin expression factor 3-like protein 1; MEF3-like protein 1; Paternally expressed gene 10 protein; Retrotransposon gag domain-containing protein 3; Retrotransposon-derived gag-like polyprotein; Ty3/Gypsy-like protein
Protein Family:Retrotransposon-derived protein
UniProt Gene Name:Peg10
UniProt Entry Name:PEG10_MOUSE
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.

Related Products