NanoPro 1000

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Important Note:

BCF stock:

* Bicine/Chaps Lysis Buffer (20mM Bicine, pH7.6, 0.6% CHAPS)

* RIPA Lysis Buffer (20mM HEPES, pH7.5, 150mM NaCl, 1% NP40 alternative, 0.25% Sodium Deoxycholate, 10% Glycerol)

* Aqueous Inhibitor Mix

* DMSO Inhibitor Mix

* HRP-secondary antibody (anti-mouse, anti-rabbit, anti-goat or anti-human
)


 

 

title Introduction

NanoProThe NanoPro 1000 system uses capillary isoelectric focusing (cIEF) separation to resolve various modification states of signaling proteins, especially phosphorylation states. Samples as few as 400nl is injected, separated, immobilized, detected and quantitated in capillaries automatically. Because of its sensitivity, characterization of signaling proteins may be invetigated directly in scarce samples, such as fine-needle tumor aspirates or primary cells.

For more detail, please visit NanoPro Technology website.

title Applications

The capillary-based immunoassay format is extremely sensitive, enabling the accurate detection and measurement of even protein isoforms.

NanoPro 1000 is capable for the following studies:

  • Oncoprotein Activation Study
  • Quantification of individual post-translational modifications, ¬†including deamidation, C-terminal lysines, glutathionylation, sialylation, oxidation, glycoslyation glycation and any other changes that will result in a pI shift.
  • Detailed phospho-isoform profiling
  • Discovery of kinase inhibitor drug effects
  • Established assays are 4EBP1, 4EBP2, ABCA1, Acetylated Lysine, AKT, AKT(p), AKT1, AKT2, AKT3, ALAS1 (Control), Anti-goat-IgG-HRP, Anti-human-IgG-HRP, ASNS, Beta actin, Beta amyloid, Beta-2-microglobulin (Control), Bim, Caspase 3, cIAP, cIAP1, cMyc, c-Myc epitope tag, COX IV, cRAF(p), CRKL, Crk-L, CRKL(p), eEF2, eEF2(p), EGFR, EGFR(p), ERK 1/2(p), ERK1, ERK1/2, ERK1/2, ERK2, ERp44, FGFR1, FGFR1(p), FLAG, FLAG epitope tag, FOXO3a, FOXO3a(p), GAPDH (Control), GFP, GluR1(p), GRASP55, GSK3-alpha (basic protein), GSK3-alpha (basic protein)(p), GSK3-alpha/beta, GSK3-beta (basic protein), GSK3-beta (basic protein)(p), GSK3¬£\, GSK3¬£], HA epitope tag, HA tag, HGF, Histone, Histone H2A, Histone H2A, K5-Acetyl, Histone H2AX, S139-Phospho, Histone H3, HMGB1, Hsc70 (Hsp73), HSP27, HSP70, HSP70 (Control), HSP90, Human Growth Hormone (hGH), IkB, IkB alpha, IkB(p), IKKalpha, IKKalpha/beta(p), IKKbeta, IKKepsilon, IKKepsilon(p), IKKgamma, JNK, JNK(p), Jun(p), LDH, LDHA, LDHB, LMP2, LRP6, Mcl-1, MEK 1/2, MEK1, MEK1(p), MEK1/2(p), MEK2, MEK2(p), MET, MET(p), Methylated Arginine, Mfn 1, Mfn 2, MFN1, MFN2, MK2, MK3, MYC, NDRG1, NDRG1(p), NF-kB, NF-kB(p), p27, p27(p), p38(p), p-4E-BP1, p53, p53(p), p70S6 kinase, PAK1, PAK1(p), PAK1,2,3, PAK1/2, PAK2, PAK3, pEGFR, Phospho-p27/KIP1, Phospho-p38, Phospho-Tyrosine (not specific to EGFR), p-JNK, PKC alpha, PKC delta, PKC delta(p), PLC gamma 1, PLC gamma 1(p), PLC-gamma 1, PRAS40, PSA, p-Stat3, p-Stat5, PTEN, PTEN(p), PTPRC/CD45, RPA2, RPA2(p), S6 Ribosomal Protein, S6 Ribosomal Protein(p), Src, Src(p), STAT1, STAT1(p), STAT3, STAT3(p), STAT5, STAT5(p), Survivin, Thioredoxin 1 (Control), Tubulin alpha, Ubiquitin and VEGF.
  • title Suggested Reading:

    NanoPro 1000 System User Guide
    NanoPro Assay Development Guide
    O'Neill RA, 2006, Isoelectric focusing technology quantifies protein signaling in 25 cells, PNAS, 103, 16153-16158.
    Fan AC, 2009, Nanofluidic proteomic assay for serial analysis of oncoprotein activation in clinical specimens, Nature Medicine, 15, 566-571.
    There are more citation may be found in ProteinSimple webpages.