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Human Natural Antibodies Recognizing
Human Natural Antibodies Recognizing Glycan Galβ1-3GlcNAc (Le C)
The level of human natural antibodies of immunoglobulin M isotype against LeC in patients with breast cancer is lower than in healthy women. The epitope specificity of these antibodies has been characterized using a printed glycan array and enzyme-linked immunosorbent assay (ELISA), the antibodies being isolated from donors’ blood using LeC-Sepharose (LeC is Galβ1-3GlcNAcβ).
The isolated antibodies recognize the disaccharide but do not bind to glycans terminated with LeC, which implies the impossibility of binding to regular glycoproteins of non-malignant cells. The avidity (as dissociation constant value) of antibodies probed with a multivalent disaccharide is 10-9 M; the nanomolar level indicates that the concentration is sufficient for physiological binding to the cognate antigen.
Testing of several breast cancer cell lines showed the strongest binding to ZR 75-1. Interestingly, only 7% of the cells were positive in a monolayer with a low density, increasing up to 96% at highest density.
The enhanced interaction (instead of the expected inhibition) of antibodies with ZR 75-1 cells in the presence of Galβ1-3GlcNAcβ disaccharide, indicates that the target epitope of anti-LeC antibodies is a molecular pattern with a carbohydrate constituent rather than a glycan.
Description: Quantitativesandwich ELISA kit for measuring Human Resistin in samples from serum, plasma, tissue homogenates. A new trial version of the kit, which allows you to test the kit in your application at a reasonable price.
Description: Quantitativesandwich ELISA kit for measuring Human Resistin in samples from serum, plasma, tissue homogenates. Now available in a cost efficient pack of 5 plates of 96 wells each, conveniently packed along with the other reagents in 5 separate kits.
Description: A competitive ELISA for quantitative measurement of Human Resistin in samples from blood, plasma, serum, cell culture supernatant and other biological fluids. This is a high quality ELISA kit developped for optimal performance with samples from the particular species.
Description: A competitive ELISA for quantitative measurement of Human Resistin in samples from blood, plasma, serum, cell culture supernatant and other biological fluids. This is a high quality ELISA kit developped for optimal performance with samples from the particular species.
Description: A competitive ELISA for quantitative measurement of Human Resistin in samples from blood, plasma, serum, cell culture supernatant and other biological fluids. This is a high quality ELISA kit developped for optimal performance with samples from the particular species.
Description: Resistin, also known as adipose tissue-specific secretory factor(ADSF) or C/EBP-epsilon-regulated myeloid-specific secreted cysteine-rich protein(XCP1) is a cysteine-rich protein that in humans is encoded by the RETN gene. The resistin gene comprises 4 exons, the first of which is untranslated, and spans approximately 1,750 bp. The human resistin gene is localized to a cloned fragment of human chromosome 19. In primates, pigs and dogs, resistin is secreted by immune and epithelial cells while in rodents, it is secreted by adipose tissue. Resistin is a cytokine whose physiologic role has been the subject of much controversy regarding its involvement with obesity and type II diabetes mellitus.
Description: Resistin, also known as adipose tissue-specific secretory factor (ADSF) or C/EBP-epsilon-regulated myeloid-specific secreted cysteine-rich protein (XCP1) is a cysteine-rich protein that in humans is encoded by the RETN gene. The resistin gene comprises 4 exons, the first of which is untranslated, and spans approximately 1,750 bp. The human resistin gene is localized to a cloned fragment of human chromosome 19. In primates, pigs and dogs, resistin is secreted by immune and epithelial cells while in rodents, it is secreted by adipose tissue. Resistin is a cytokine whose physiologic role has been the subject of much controversy regarding its involvement with obesity and type II diabetes mellitus.
RETN gene belongs to the family defined by the mouse resistin-like genes. The characteristic feature of this family is the C-terminal stretch of 10 cys residues with identical spacing. The mouse homolog of this protein is secreted by adipocytes, and
Description: Quantitative sandwich ELISA for measuring Human Resistin in samples from cell culture supernatants, serum, whole blood, plasma and other biological fluids.
RETN gene belongs to the family defined by the mouse resistin-like genes. The characteristic feature of this family is the C-terminal stretch of 10 cys residues with identical spacing. The mouse homolog of this protein is secreted by adipocytes, and
Description: Quantitative sandwich ELISA for measuring Human Resistin in samples from cell culture supernatants, serum, whole blood, plasma and other biological fluids.
RETN gene belongs to the family defined by the mouse resistin-like genes. The characteristic feature of this family is the C-terminal stretch of 10 cys residues with identical spacing. The mouse homolog of this protein is secreted by adipocytes, and
Description: Quantitative sandwich ELISA for measuring Human Resistin in samples from cell culture supernatants, serum, whole blood, plasma and other biological fluids.
Description: Enzyme-linked immunosorbent assay kit for quantification of Human Resistin in samples from serum, plasma, tissue homogenates and other biological fluids.
Description: Human RETN ELISA development kit contains the key components required for the quantitative measurement of natural and/or recombinant RETN in a sandwich ELISA format within the range of 16-2,000 pg/mL. Using the ELISA protocol described below, this kit provides sufficient reagents to assay RETN in approximately 1,500 ELISA plate wells.
Description: This assay is a sandwich Enzyme Linked-Immuno-Sorbent Assay (ELISA). It is developed for quantitative measurement of Human RETN in serum, plasma and other biological fluids.
Should the Human Resistin (RETN) ELISA Kit is proven to show malperformance, you will receive a refund or a free replacement.
Description: A sandwich quantitative ELISA assay kit for detection of Human Resistin (RETN) in samples from serum, plasma, tissue homogenates, cell lysates, cell culture supernates or other biological fluids.
Should the Human Resistin (RETN) ELISA Kit is proven to show malperformance, you will receive a refund or a free replacement.
Description: A sandwich quantitative ELISA assay kit for detection of Human Resistin (RETN) in samples from serum, plasma, tissue homogenates, cell lysates, cell culture supernates or other biological fluids.
Targeted bottom-up characterization of recombinant monoclonal antibodies by multi-dimensional LC/MS
On-line bottom-up approaches have recently emerged as promising alternatives to standard off-line processes for char-acterizing post-translational modifications (PTMs) of therapeutic monoclonal antibodies (mAbs).
The benefits of on-line processing include reductions in required sample amount and sample handling, as well as reducing the overall turnaround time. However, shortening digestion time for the on-line approach of an intact mAb can cause incomplete peptide cleavages, leading to low sequence coverage and poor repeatability of analyses.
For the first time, we describe a novel, automated targeted bottom-up strategy consisting of reducing the complexity of intact mAb by digesting the product into small ~25 kDa fragments, followed by an on-line peptide mapping analysis of each fragment. For this pur-pose, a 4D-LC/MS method was developed using a immobilized IdeS-HPLC column as a first dimension (1D) for on-line digestion, followed by a (2D) on-column reversed-phase liquid chromatography (RPLC) for reduction and fragments separation.
Then, only one fragment was selected for digestion using a (3D) immobilized trypsin cartridge and finally, the obtained peptides were analyzed by (4D) RPLC-MS. This strategy considerably improved the on-line digestion effi-ciency with higher sequence coverages (LC and HC > 97%), thus allowing various PTMs including oxidation, deami-dation, and isomerization located in the complementarity-determining regions (CDRs), as well as N-glycans present on the Fc/2 fragment, to be monitored with similar sensitivity to those obtained with standard off-line approaches.
Additional investigations at a middle-up level were also performed via a 3D-LC/MS approach within the same system, demonstrating the feasibility to achieve a multi-level comprehensive characterization of mAbs.
Monophosphoryl lipid A-induced activation of plasmacytoid dendritic cells enhances the anti-cancer effects of anti-PD-L1 antibodies
Monophosphoryl lipid A (MPLA) is a toll-like receptor 4 ligand that promotes immune activation in mice and humans, without undesired inflammation. Immunotherapy by the combining immune checkpoint blockade and MPLA has shown promising anti-cancer effects in both mice and humans.
In this study, we explored how MPLA enhanced the anti-cancer effects of anti-PD-L1 antibodies (Abs). Anti-cancer immunity induced by the combination of anti-PD-L1 Abs and MPLA failed in CD4 and CD8 cell-depleted mice. Moreover, the combination treatment of anti-PD-L1 Abs and MPLA synergistically enhanced the activation of plasmacytoid dendritic cells (pDCs) in the mouse in vivo, while conventional DCs were not. In addition, mice treated with anti-PD-L1 Abs and MPLA were not protected from B16 melanoma by blockade of interferon-alpha receptor (IFNAR).
The combination of anti-PD-L1 Abs and MPLA also promoted human peripheral blood pDC activation and induced IFN-α-dependent T cell activation. Therefore, these results demonstrate that MPLA enhances anti-PD-L1 Ab-mediated anti-cancer immunity through the activation and IFN-α production of pDCs.