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Mouse Proprotein Convertase 9/PCSK9 Quantikine ELISA Kit

Catalog #: MPC900
30 Citations 8 Reviews Datasheet / COA / SDS
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Summary Sample Values Precision Recovery Linearity Data Examples Product Datasheets Preparation and Storage Background Related Research Areas

Mouse Proprotein Convertase 9/PCSK9 Quantikine ELISA Kit Summary

Assay Type
Solid Phase Sandwich ELISA
Format
96-well strip plate
Assay Length
4.5 hours
Sample Type & Volume Required Per Well
Cell Culture Supernates (50 uL), Cell Lysates (10 uL), Serum (10 uL), EDTA Plasma (10 uL), Heparin Plasma (10 uL)
Sensitivity
21.9 pg/mL
Assay Range
62.5 - 4,000 pg/mL (Cell Culture Supernates, Cell Lysates, Serum, EDTA Plasma, Heparin Plasma)
Specificity
Natural and recombinant mouse PCSK9. This kit detects 60 kDa, 53 kDa, and LDLR-complexed recombinant mouse PCSK9.
Cross-reactivity
< 0.5% cross-reactivity observed with available related molecules.< 50% cross-species reactivity observed with species tested.
Interference
No significant interference observed with available related molecules.

Sample Values

Sample TypeMean (ng/mL)Range (ng/mL)Standard Deviation (ng/mL)
Serum (n=20)343153-649163
EDTA plasma (n=20)337166-612118
Heparin plasma (n=20)324141-565100

Cell Culture Supernates - Organs from 2-3 mice were chopped into 1-2 mm pieces and cultured in RPMI supplemented with 10% fetal bovine serum, 2 mM L-glutamine, 100 U/mL penicillin, and 100 mg/mL streptomycin sulfate. The cell culture supernates were unstimulated or stimulated with 1.0 mg/mL of lipopolysaccharide for 1 or 2 days. An aliquot of the cell culture supernate was removed and assayed for levels of mouse PCSK9. Cell culture supernates from mouse brain, heart, lung, and spleen tissue measured below the low standard, 62.5 pg/mL. Cell culture supernates from liver and kidney tissue measured as follows.

Tissue Type(pg/mL)
Kidney (unstimulated for 1 day)140
Liver (unstimulated for 2 days)889
Liver (stimulated with 1.0 mg/mL LPS for 2 days)899

Tissue Lysates - Organs from 2-3 mice were rinsed with PBS to remove excess blood, chopped into 1-2 mm pieces, homogenized with a tissue homogenizer, and 1% v/v Triton™ X-100 was added. An aliquot of each tissue lysate was removed and assayed for levels of mouse PCSK9.

Tissue Type(pg/mL)
Brain1741
Heart1603
Kidney8953
Liver36,350
Lung3898
Spleen1800


Product Summary

The Quantikine Mouse PCSK9 Immunoassay is a 4.5 hour solid-phase ELISA designed to measure mouse PCSK9 in cell culture supernates, cell lysates, serum, and plasma. It contains NS0-expressed recombinant mouse PCSK9 and antibodies raised against the recombinant factor. This immunoassay has been shown to accurately quantitate the recombinant factor. Results obtained using natural mouse PCSK9 showed linear curves that were parallel to the standard curves obtained using the Quantikine kit standards. These results indicate that this kit can be used to determine relative mass values for naturally occurring mouse PCSK9.

Precision

Inter-Assay Precision (Precision between assays) Three samples of known concentration were tested in separate assays to assess inter-assay precision

Cell Culture Supernates, Cell Lysates, Serum, EDTA Plasma, Heparin Plasma

Intra-Assay Precision Inter-Assay Precision
Sample 2 3
n 40 40
Mean (pg/mL) 469 1511
Standard Deviation 25.9 102
CV% 5.5 6.8

Recovery

The recovery of mouse PCSK9 spiked to three levels throughout the range of the assay in various matrices was evaluated.

Sample Type Average % Recovery Range %
Cell Culture Supernates (n=6) 102 94-115
Cell Lysates (n=5) 103 91-117

Linearity

To assess the linearity of the assay, samples containing and/or spiked with high concentrations of mouse PCSK9 in each matrix were diluted with Calibrator Diluent and then assayed.
Mouse Proprotein Convertase 9/PCSK9 ELISA Linearity

Scientific Data

N/A Mouse Proprotein Convertase 9/PCSK9 ELISA Standard Curve View Larger

Mouse Proprotein Convertase 9/PCSK9 ELISA Standard Curve

Product Datasheets

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Preparation and Storage

Shipping
The product is shipped at ambient temperature. Upon receipt, store it immediately at the temperature recommended below.
Storage
Store the unopened product at 2 - 8 °C. Do not use past expiration date.

Background: Proprotein Convertase 9/PCSK9

PCSK9 (proprotein convertase subtilisin kexin 9), also called proprotein convertase 9 or NARC-1 (neural apoptosis-regulated convertase 1), is a member of the proteinase K subfamily of subtilisinrelated serine endoproteases. Mouse PCSK9 cDNA encodes 694 amino acids, including a signal peptide, a prodomain, and a catalytic domain. PCSK9 is highly expressed in the liver, intestine, and kidney. It is initially synthesized as a soluble 74 kDa precursor protein. In the endoplasmic reticulum, it undergoes autocatalytic intramolecular cleavage to generate a 14 kDa prodomain and a 60 kDa catalytic domain. While within the secretion pathway, the prodomain remains associated and functions as a chaperone for the catalytic domain (1-4). During secretion, a portion of active PCSK9 may undergo additional N-terminal proteolysis by furin or proprotein convertase 5/6A, creating an inactive 53 kDa form (5). This cleavage site is conserved between mouse and human or rat PCSK9, which share 78% or 93% amino acid sequence identity, respectively. While the 60 kDa protein is the major form, its ratio with the 53 kDa forms is variable in humans (5, 6). 

The primary physiologic function of PCSK9 is to mediate the degradation of low density lipoprotein receptor (LDLR). Early observations indicated that gain-of-function missense mutations in the human PCSK9 gene can cause an autosomal dominant form of hypercholesterolemia (7, 8). The expression of PCSK9 is also upregulated by the sterol regulatory element binding proteins (SREBPs), a family of transcription factors that are responsible for the upregulation of genes involved in cholesterol and fatty acid metabolism, such as the LDLR gene (9, 10). Further experimental evidence revealed that when the mouse PCSK9 gene is deleted, LDLR expression in hepatocytes is increased. Conversely, PCSK9 over-expression decreases liver LDLR protein expression (11, 12). In humans, genetic analyses have shown that individuals who have nonsense or loss-of-function mutations in the PCSK9 gene have significantly lower plasma LDL cholesterol levels, while in mouse, administration of a PCSK9 neutralizing antibody or antisense oligonucleotides lowers serum cholesterol (1, 13-15). These investigations clearly indicate that PCSK9 plays a key role in reducing the hepatic LDLR levels. Paradoxically, administration of cholesterol-lowering drugs such as statins appear to enhance production of PCSK9 (6). 
The underlying mechanism of cholesterol regulation by PCSK9 is as follows: under normal physiologic conditions, the LDLR is internalized at the cell surface and directed to the endosomes in order to be recycled back to the cell surface. PCSK9 binds to the EGF domain of the LDLR and prevents LDLR from being sorted to the endosomes. Instead, the PCSK9/LDLR complex is redistributed to the lysosomes for degradation (16-18). As such, PCSK9 regulates the amount of LDLR in the circulation and hence, modulates cholesterol levels. Serum PCSK9 concentrations have been found to be directly associated with cholesterol levels (19, 20). Since PCSK9 loss-of-function mutations strikingly reduce risk of coronary heart diseases, PCSK9 has become an attractive drug target (1, 21, 22). One approach is to generate small molecules that are able to interfere with PCSK9 autoactivation and its interaction with LDLR. Other approaches aiming to reduce the amount of PCSK9 in the circulation, such as small interfering RNAs (siRNAs), have also shown promise (23, 24).

Entrez Gene IDs:
255738 (Human); 100102 (Mouse); 298296 (Rat); 102142788 (Cynomolgus Monkey)
Alternate Names:
EC 3.4.21; EC 3.4.21.111; FH3; FH3neural apoptosis regulated convertase 1; FHCL3; HCHOLA3; hypercholesterolemia, autosomal dominant 3; LDLCQ1; NARC1; NARC-1; NARC-1convertase subtilisin/kexin type 9 preproprotein; NARC1EC 3.4.21.-; Neural apoptosis-regulated convertase 1; PC9; PCSK9; Proprotein Convertase 9; proprotein convertase subtilisin/kexin type 9; Subtilisin/kexin-like protease PC9
&#9888; WARNING: This product can expose you to chemicals including N,N-Dimethylforamide, which is known to the State of California to cause cancer. For more information, go to www.P65Warnings.ca.gov.

Assay Procedure

Refer to the product for complete assay procedure.

Bring all reagents and samples to room temperature before use. It is recommended that all samples, standards, and controls be assayed in duplicate.
  1.   Prepare all reagents, standard dilutions, and samples as directed in the product insert.
  2.   Remove excess microplate strips from the plate frame, return them to the foil pouch containing the desiccant pack, and reseal.
    3. 50 µL Assay Diluent
  3.   Add 50 µL of Assay Diluent to each well.
    4. 50 µL Standard, Control, or Sample
  4.   Add 50 µL of Standard, Control, or sample to each well. Cover with a plate sealer, and incubate at room temperature for 2 hours.
  5.   Aspirate each well and wash, repeating the process 4 times for a total of 5 washes.
    6. 100 µL Conjugate
  6.   Add 100 µL of Conjugate to each well. Cover with a new plate sealer, and incubate at room temperature for 2 hours.
  7.   Aspirate and wash 5 times.
    8. 100 µL Substrate Solution
  8.   Add 100 µL Substrate Solution to each well. Incubate at room temperature for 30 minutes. PROTECT FROM LIGHT.
    9. 100 µL Stop Solution
  9.   Add 100 µL of Stop Solution to each well. Read at 450 nm within 30 minutes. Set wavelength correction to 540 nm or 570 nm.

Citations for Mouse Proprotein Convertase 9/PCSK9 Quantikine ELISA Kit

R&D Systems personnel manually curate a database that contains references using R&D Systems products. The data collected includes not only links to publications in PubMed, but also provides information about sample types, species, and experimental conditions.

30 Citations: Showing 1 - 10
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  1. Durable and efficient gene silencing in vivo by hit-and-run epigenome editing
    Authors: Cappelluti, MA;Mollica Poeta, V;Valsoni, S;Quarato, P;Merlin, S;Merelli, I;Lombardo, A;
    Nature
    Species: Mouse
    Sample Types: Plasma
  2. Integrated compact regulators of protein activity enable control of signaling pathways and genome-editing in vivo
    Authors: Franko, N;da Silva Santinha, AJ;Xue, S;Zhao, H;Charpin-El Hamri, G;Platt, RJ;Teixeira, AP;Fussenegger, M;
    Cell discovery
    Species: Mouse
    Sample Types: Serum
  3. Design and application of the transformer base editor in mammalian cells and mice
    Authors: Han, W;Gao, BQ;Zhu, J;He, Z;Li, J;Yang, L;Chen, J;
    Nature protocols
    Species: Transgenic Mouse
    Sample Types: Serum
  4. Continuous directed evolution of a compact CjCas9 variant with broad PAM compatibility
    Authors: Schmidheini, L;Mathis, N;Marquart, KF;Rothgangl, T;Kissling, L;Böck, D;Chanez, C;Wang, JP;Jinek, M;Schwank, G;
    Nature chemical biology
    Species: Mouse
    Sample Types: Tissue Homogenates
  5. A Type II-B Cas9 nuclease with minimized off-targets and reduced chromosomal translocations in vivo
    Authors: Bestas, B;Wimberger, S;Degtev, D;Madsen, A;Rottner, AK;Karlsson, F;Naumenko, S;Callahan, M;Touza, JL;Francescatto, M;Möller, CI;Badertscher, L;Li, S;Cerboni, S;Selfjord, N;Ericson, E;Gordon, E;Firth, M;Chylinski, K;Taheri-Ghahfarokhi, A;Bohlooly-Y, M;Snowden, M;Pangalos, M;Nuttall, B;Akcakaya, P;Sienski, G;Maresca, M;
    Nature communications
    Species: Mouse
    Sample Types: Plasma
  6. High-fidelity Cas13 variants for targeted RNA degradation with minimal collateral effects
    Authors: Tong, H;Huang, J;Xiao, Q;He, B;Dong, X;Liu, Y;Yang, X;Han, D;Wang, Z;Wang, X;Ying, W;Zhang, R;Wei, Y;Xu, C;Zhou, Y;Li, Y;Cai, M;Wang, Q;Xue, M;Li, G;Fang, K;Zhang, H;Yang, H;
    Nature biotechnology
    Species: Mouse
    Sample Types: Serum
  7. A cVLP-Based Vaccine Displaying Full-Length PCSK9 Elicits a Higher Reduction in Plasma PCSK9 Than Similar Peptide-Based cVLP Vaccines
    Authors: L Goksøyr, M Skrzypczak, M Sampson, MA Nielsen, A Salanti, TG Theander, AT Remaley, WA De Jongh, AF Sander
    Vaccines, 2022-12-20;11(1):.
    Species: Mouse
    Sample Types: Plasma
  8. Rosa26-LSL-dCas9-VPR: a versatile mouse model for tissue specific and simultaneous activation of multiple genes for drug discovery
    Authors: D Pakalnišky, T Schönberge, B Strobel, B Stierstorf, T Lamla, M Schuler, M Lenter
    Scientific Reports, 2022-11-10;12(1):19268.
    Species: Transgenic Mouse
    Sample Types: Plasma
  9. A multienzyme S-nitrosylation cascade regulates cholesterol homeostasis
    Authors: CT Stomberski, NM Venetos, HL Zhou, Z Qian, BR Collison, SJ Field, RT Premont, JS Stamler
    Cell Reports, 2022-10-25;41(4):111538.
    Species: Mouse
    Sample Types: Serum
  10. Diverse Effects of Cilostazol on Proprotein Convertase Subtilisin/Kexin Type 9 between Obesity and Non-Obesity
    Authors: PW Chen, SY Tseng, HY Chang, CH Lee, TH Chao
    International Journal of Molecular Sciences, 2022-08-29;23(17):.
    Species: Mouse
    Sample Types: Serum
  11. Caffeine blocks SREBP2-induced hepatic PCSK9 expression to enhance LDLR-mediated cholesterol clearance
    Authors: PF Lebeau, JH Byun, K Platko, P Saliba, M Sguazzin, ME MacDonald, G Paré, GR Steinberg, LJ Janssen, SA Igdoura, MA Tarnopolsk, SR Wayne Chen, NG Seidah, J Magolan, RC Austin
    Nature Communications, 2022-02-09;13(1):770.
    Species: Mouse
    Sample Types: Cell Culture Supernates
  12. Engineered red blood cells carrying PCSK9 inhibitors persistently lower LDL and prevent obesity
    Authors: R Deshycka, V Sudaryo, NJ Huang, Y Xie, LY Smeding, MK Choi, HL Ploegh, HF Lodish, N Pishesha
    PLoS ONE, 2021-11-03;16(11):e0259353.
    Species: Mouse
    Sample Types: Plasma
  13. In vivo adenine base editing of PCSK9 in macaques reduces LDL cholesterol levels
    Authors: T Rothgangl, MK Dennis, PJC Lin, R Oka, D Witzigmann, L Villiger, W Qi, M Hruzova, L Kissling, D Lenggenhag, C Borrelli, S Egli, N Frey, N Bakker, JA Walker, AP Kadina, DV Victorov, M Pacesa, S Kreutzer, Z Kontarakis, A Moor, M Jinek, D Weissman, M Stoffel, R van Boxtel, K Holden, N Pardi, B Thöny, J Häberle, YK Tam, SC Semple, G Schwank
    Nature Biotechnology, 2021-05-19;0(0):.
    Species: Mouse
    Sample Types: Cell Culture Supernates
  14. Eliminating base-editor-induced genome-wide and transcriptome-wide off-target mutations
    Authors: L Wang, W Xue, H Zhang, R Gao, H Qiu, J Wei, L Zhou, YN Lei, X Wu, X Li, C Liu, J Wu, Q Chen, H Ma, X Huang, C Cai, Y Zhang, B Yang, H Yin, L Yang, J Chen
    Nature Cell Biology, 2021-05-10;23(5):552-563.
    Species: Mouse
    Sample Types: Serum
  15. Genetic deletion of Abcc6 disturbs cholesterol homeostasis in mice
    Authors: B Ibold, J Tiemann, I Faust, U Ceglarek, J Dittrich, TGMF Gorgels, AAB Bergen, O Vanakker, M Van Gils, C Knabbe, D Hendig
    Scientific Reports, 2021-01-22;11(1):2137.
    Species: Mouse
    Sample Types: Serum
  16. Optimization of S.�aureus dCas9 and CRISPRi Elements for a Single Adeno-Associated Virus that Targets an Endogenous Gene
    Authors: JR Backstrom, J Sheng, MC Wang, A Bernardo-C, TS Rex
    Mol Ther Methods Clin Dev, 2020-09-06;19(0):139-148.
    Species: Mouse
    Sample Types: Cell Culture Supernates
  17. Synthetic immunomodulation with a CRISPR super-repressor in vivo
    Authors: F Moghadam, R LeGraw, JJ Velazquez, NC Yeo, C Xu, J Park, A Chavez, MR Ebrahimkha, S Kiani
    Nat. Cell Biol., 2020-09-03;22(9):1143-1154.
    Species: Mouse
    Sample Types: Plasma
  18. In vivo genome and base editing of a human PCSK9 knock-in hypercholesterolemic mouse model
    Authors: A Carreras, LS Pane, R Nitsch, K Madeyski-B, M Porritt, P Akcakaya, A Taheri-Gha, E Ericson, M Bjursell, M Perez-Alca, F Seeliger, M Althage, R Knöll, R Hicks, LM Mayr, R Perkins, D Lindén, J Borén, M Bohlooly-Y, M Maresca
    BMC Biol., 2019-01-15;17(1):4.
    Species: Mouse
    Sample Types: Plasma
  19. In utero CRISPR-mediated therapeutic editing of metabolic genes
    Authors: AC Rossidis, JD Stratigis, AC Chadwick, HA Hartman, NJ Ahn, H Li, K Singh, BE Coons, L Li, W Lv, PW Zoltick, D Alapati, W Zacharias, R Jain, EE Morrisey, K Musunuru, WH Peranteau
    Nat. Med., 2018-10-08;24(10):1513-1518.
    Species: Mouse
    Sample Types: Plasma
  20. In vivo CRISPR editing with no detectable genome-wide off-target mutations
    Authors: P Akcakaya, ML Bobbin, JA Guo, J Malagon-Lo, K Clement, SP Garcia, MD Fellows, MJ Porritt, MA Firth, A Carreras, T Baccega, F Seeliger, M Bjursell, SQ Tsai, NT Nguyen, R Nitsch, LM Mayr, L Pinello, M Bohlooly-Y, MJ Aryee, M Maresca, JK Joung
    Nature, 2018-09-12;0(0):.
    Species: Mouse
    Sample Types: Plasma
  21. RNA-guided transcriptional silencing in vivo with S. aureus CRISPR-Cas9 repressors
    Authors: PI Thakore, JB Kwon, CE Nelson, DC Rouse, MP Gemberling, ML Oliver, CA Gersbach
    Nat Commun, 2018-04-26;9(1):1674.
    Species: Mouse
    Sample Types: Serum
  22. Reduced Blood Lipid Levels With In Vivo CRISPR-Cas9 Base Editing of ANGPTL3
    Authors: AC Chadwick, NH Evitt, W Lv, K Musunuru
    Circulation, 2018-02-27;137(9):975-977.
    Species: Mouse
    Sample Types: Plasma
  23. Generation and Characterization of a Novel Small Biologic Alternative to Proprotein Convertase Subtilisin/Kexin Type 9 (PCSK9) Antibodies, DS-9001a, Albumin Binding Domain-Fused Anticalin Protein
    Authors: Y Masuda, S Yamaguchi, C Suzuki, T Aburatani, Y Nagano, R Miyauchi, E Suzuki, N Yamamura, K Nagatomo, H Ishihara, K Okuno, F Nara, G Matschiner, R Hashimoto, T Takahashi, T Nishizawa
    J. Pharmacol. Exp. Ther., 2018-02-20;365(2):368-378.
    Species: Mouse
    Sample Types: Plasma
  24. Development of vaccine for dyslipidemia targeted to a proprotein convertase subtilisin/kexin type 9 (PCSK9) epitope in mice
    Authors: R Kawakami, Y Nozato, H Nakagami, Y Ikeda, M Shimamura, S Yoshida, J Sun, T Kawano, Y Takami, T Noma, H Rakugi, T Minamino, R Morishita
    PLoS ONE, 2018-02-13;13(2):e0191895.
    Species: Mouse
    Sample Types: Plasma
  25. Dynamin-Related Protein 1 Inhibition Attenuates Cardiovascular Calcification in the Presence of Oxidative Stress
    Authors: M Rogers, N Maldonado-, JD Hutcheson, C Goettsch, S Goto, I Yamada, T Faits, H Sesaki, M Aikawa, E Aikawa
    Circ. Res., 2017-06-12;0(0):.
    Species: Mouse
    Sample Types: Serum
  26. Regulation of lipid metabolism by obeticholic acid in hyperlipidemic hamsters
    Authors: Bin Dong
    J. Lipid Res, 2016-12-09;0(0):.
    Species: Hamster
    Sample Types: Serum
  27. Endoplasmic reticulum stress and Ca2+ depletion differentially modulate the sterol-regulatory protein PCSK9 to control lipid metabolism
    Authors: Paul Lebeau
    J. Biol. Chem, 2016-12-01;0(0):.
    Species: Mouse
    Sample Types: Plasma
  28. Reduction of circulating PCSK9 and LDL-C levels by liver-specific knockdown of HNF1alpha in normolipidemic mice.
    Authors: Shende V, Wu M, Singh A, Dong B, Kan C, Liu J
    J Lipid Res, 2015-02-04;56(4):801-9.
    Species: Mouse
    Sample Types: Serum
  29. Inhibition of PCSK9 transcription by berberine involves down-regulation of hepatic HNF1alpha protein expression through the ubiquitin-proteasome degradation pathway.
    Authors: Dong B, Li H, Singh A, Cao A, Liu J
    J Biol Chem, 2014-12-24;290(7):4047-58.
    Species: Hamster, Mouse
    Sample Types: Serum
  30. Permanent alteration of PCSK9 with in vivo CRISPR-Cas9 genome editing.
    Authors: Ding Q, Strong A, Patel K, Ng S, Gosis B, Regan S, Cowan C, Rader D, Musunuru K
    Circ Res, 2014-06-10;115(5):488-92.
    Species: Mouse
    Sample Types: Plasma

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Mouse Proprotein Convertase 9/PCSK9 Quantikine ELISA Kit
By Anonymous on 10/05/2023
Sample Tested: mouse hepatocyte
Mouse Proprotein Convertase 9/PCSK9 Quantikine ELISA Kit MPC900
Mouse Proprotein Convertase 9/PCSK9 Quantikine ELISA Kit
By Anonymous on 05/19/2023
Sample Tested: EDTA Plasma,Serum
Mouse Proprotein Convertase 9/PCSK9 Quantikine ELISA Kit MPC900
Mouse Proprotein Convertase 9/PCSK9 Quantikine ELISA Kit
By Anonymous on 03/15/2023
Sample Tested: Serum
Mouse Proprotein Convertase 9/PCSK9 Quantikine ELISA Kit MPC900
Mouse Proprotein Convertase 9/PCSK9 Quantikine ELISA Kit
By Anonymous on 02/08/2022
Sample Tested: Serum

stable kit

Mouse Proprotein Convertase 9/PCSK9 Quantikine ELISA Kit MPC900
Mouse Proprotein Convertase 9/PCSK9 Quantikine ELISA Kit
By Anonymous on 03/22/2020
Sample Tested: 2120Ep human embryonal carcinoma cell line,3T3-L1 mouse embryonic fibroblast adipose-like cell line
Mouse Proprotein Convertase 9/PCSK9 Quantikine ELISA Kit
By Anonymous on 10/15/2018
Sample Tested: Heparin Plasma
Mouse Proprotein Convertase 9/PCSK9 Quantikine ELISA Kit MPC900
Mouse Proprotein Convertase 9/PCSK9 Quantikine ELISA Kit
By Anonymous on 08/29/2018
Sample Tested: Serum
Mouse Proprotein Convertase 9/PCSK9 Quantikine ELISA Kit
By Adrianne Pittman on 04/26/2017
Sample Tested: Mouse spleenocytes