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Mouse Phosphoglycerate mutase 2 ELISA Kit

SKU:
MOFI00472
Product Type:
ELISA Kit
Size:
96 Assays
Uniprot:
O70250
Sensitivity:
46.875pg/ml
Range:
78.125-5000pg/ml
ELISA Type:
Sandwich
Synonyms:
Pgam2, PhosphoglyceRate mutase isozyme M, PGAM-M, BPG-dependent PGAM 2, Muscle-specific phosphoglyceRate mutase,
Reactivity:
Mouse
Research Area:
Metabolism
€649
Frequently bought together:

Description

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Mouse Phosphoglycerate mutase 2 ELISA Kit

The Mouse Phosphoglycerate Mutase 2 ELISA Kit is specifically designed for the quantitative detection of phosphoglycerate mutase 2 in mouse serum, plasma, and cell lysates. This kit offers high sensitivity and specificity, ensuring accurate and reproducible results for a variety of research applications.Phosphoglycerate mutase 2 is an enzyme that plays a crucial role in glycolysis and gluconeogenesis, making it essential for energy production in cells. It is also involved in various metabolic pathways and has been implicated in conditions such as cancer and metabolic disorders.

Therefore, the measurement of phosphoglycerate mutase 2 levels can provide valuable insights into disease mechanisms and potential therapeutic targets.Overall, the Mouse Phosphoglycerate Mutase 2 ELISA Kit is a reliable tool for researchers studying metabolism, cell biology, and disease pathways in mouse models. Its high performance and accuracy make it a valuable asset in the laboratory for advancing scientific discoveries and drug development.

Product Name:Mouse Phosphoglycerate mutase 2 ELISA Kit
Product Code:MOFI00472
Size:96 Assays
Alias:Pgam2, PhosphoglyceRate mutase isozyme M, PGAM-M, BPG-dependent PGAM 2, Muscle-specific phosphoglyceRate mutase
Detection Method:Sandwich ELISA
Application:This immunoassay kit allows for the in vitro quantitative determination of Mouse Pgam2 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 Mouse Pgam2 and the recovery rates were calculated by comparing the measured value to the expected amount of Mouse Pgam2 in samples.
MatrixRecovery range(%)Average(%)
serum(n=5)87-10195
EDTA plasma(n=5)85-10291
UFH plasma(n=5)89-10498
Linearity:The linearity of the kit was assayed by testing samples spiked with appropriate concentration of Mouse Pgam2 and their serial dilutions. The results were demonstrated by the percentage of calculated concentration to the expected.
Sample1:21:41:8
serum(n=5)85-94%86-103%86-90%
EDTA plasma(n=5)85-100%86-95%84-99%
UFH plasma(n=5)84-100%83-100%81-94%
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
UniprotO70250
UniProt Protein Function:PGAM2: Interconversion of 3- and 2-phosphoglycerate with 2,3- bisphosphoglycerate as the primer of the reaction. Can also catalyze the reaction of EC 5.4.2.4 (synthase) and EC 3.1.3.13 (phosphatase), but with a reduced activity. Defects in PGAM2 are the cause of glycogen storage disease type 10 (GSD10). A metabolic disorder characterized by myoglobinuria, increased serum creatine kinase levels, decreased phosphoglycerate mutase activity, myalgia, muscle pain, muscle cramps and excercise intolerance. Belongs to the phosphoglycerate mutase family. BPG- dependent PGAM subfamily.
UniProt Protein Details:

Protein type:Carbohydrate Metabolism - glycolysis and gluconeogenesis; EC 3.1.3.13; EC 5.4.2.11; EC 5.4.2.4; Isomerase; Phosphatase (non-protein)

Chromosomal Location of Human Ortholog: 11|11 A1

Cellular Component: cytosol; nucleus

Molecular Function:2,3-bisphosphoglycerate-dependent phosphoglycerate mutase activity; bisphosphoglycerate mutase activity; catalytic activity; cofactor binding; hydrolase activity; intramolecular transferase activity, phosphotransferases; isomerase activity; phosphoglycerate mutase activity; protein binding

Biological Process: gluconeogenesis; glycolytic process; metabolic process; Notch signaling pathway; regulation of pentose-phosphate shunt; response to inorganic substance; response to mercury ion; striated muscle contraction

UniProt Code:O70250
NCBI GenInfo Identifier:9256624
NCBI Gene ID:56012
NCBI Accession:NP_061358.1
UniProt Related Accession:O70250
Molecular Weight:28,827 Da
NCBI Full Name:phosphoglycerate mutase 2
NCBI Synonym Full Names:phosphoglycerate mutase 2
NCBI Official Symbol:Pgam2  
NCBI Protein Information:phosphoglycerate mutase 2
UniProt Protein Name:Phosphoglycerate mutase 2
UniProt Synonym Protein Names:BPG-dependent PGAM 2; Muscle-specific phosphoglycerate mutase; Phosphoglycerate mutase isozyme M; PGAM-M
Protein Family:Phosphoglycerate mutase
UniProt Gene Name:Pgam2  

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

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 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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