Related Categories: Application rate, Biochemicals and reagents, Enzymes, Inhibitors and substrates, Isolation of proteins and nucleic acids

Quality level: 100

Form: lyophilized

Specific activity: ≥25 units / mg protein

Storage condition:

OK to freeze
Dried

Storage conditions: -20C

General description

Elastase is native to the porcine pancreas. Serine protease that catalyzes the hydrolysis of proteins and peptides (especially at bonds adjacent to neutral amino acid residues), including albumin, casein, denatured collagen, elastin, fibrin, and hemoglobin, and of various synthetic substrates containing aspartic acid, phenylalanine, or tyrosine. Inhibited by DFP, elastin, and α2-macroglobulin.

Elastase is native to the porcine pancreas. Catalyzes the hydrolysis of proteins and peptides (especially at bonds adjacent to neutral amino acid residues), including albumin, casein, denatured collagen, elastin, fibrin, and hemoglobin, and of various synthetic substrates containing aspartic acid, glutamic acid, phenylalanine, or tyrosine. Preferably it cleaves peptide bonds at the carbonyl end of amino acid residues with small hydrophobic side chains, such as glycine, valine, leucine, isoleucine, and particularly alanine. Inhibited by DFP, elastin, and α2-macroglobulin. It has an optimal pH of 7.8-8.5; pI = 9.5.

Packaging

1000, 250 u in plastic ampoule

Warning

Toxicity: Standard handling (A)

Definition of unit

One unit is defined as the amount of enzyme that will hydrolyze 1.0 mole of Suc-Ala-Ala-Pro-Abu-PNA (catalog # 324699) per minute at 25 ° C, pH 8.0.

Physical form

Lyophilized from 50 mM trehalose, 1 mM acetic acid.

Reconstitution

After reconstitution, elastase can be stored at 4 ° C at pH 6.0 for long-term use. If incubated at room temperature at or near its optimum pH, elastase rapidly autolyzes to a mixture of peptides. Stock solutions are stable for up to 2 months at 4 ° C, pH 6.0.

General description

The product binds human IgG and not other human Ig. Immunoglobulins are composed of antigen-binding domains of two fragments (Fab) and one crystallizable fragment (Fc). The gene encoding the IgG gene cluster is found on human chromosome 14. Anti-human IgG antiserum (Fc specific) is produced in goat using purified human IgG, Fc fragment, as an immunogen. Affinity isolated antibody is obtained from goat anti-human IgG antiserum by immunospecific purification that removes essentially all goat serum proteins, including immunoglobulins, that do not specifically bind to the Fc fragment of human IgG.

Specificity

Specificity for the human IgG Fc fragment is determined by ELISA and immunoelectrophoresis (IEP). The antibody preparation is specific for human IgG, Fc fragment when tested against purified human IgA, IgG (Fc and Fab fragments), IgM, Bence Jones kappa, and Bence Jones lambda myeloma proteins. No reactivity is observed with the Fab fragment of human IgG, light chains, IgA or IgM. The affinity-purified anti-human IgG (Fc-specific) reagent offers the advantage of increased sensitivity for human IgG without cross-reactivity with other substances present on the cell membrane or surface.

The lack of cross-species cross-reactivity with mouse or rat serum proteins makes this product excellent for screening human monoclonal antibodies produced by hybridoma cells grown in vivo in mouse or rat ascites fluids. This product has the ability to detect all subclasses of human IgG in human biological fluids or tissues from normal or pathological situations such as cancer or autoimmune diseases. It is effective as a second antibody reagent in immunoassay procedures and can be used as a starting material for conjugates using enzymes or fluorescent dyes.

Immunogen

Anti-human IgG antiserum (Fc specific) is produced in goats using purified human IgG, Fc fragment, as an immunogen

Request

The anti-human IgG antibody (specific for Fc) produced in goats has been used:

• in double capture ELISA to measure antiglobulin responses in the serum of transplant patients treated with monoclonal antibodies CD52 (CAMPATH-1G)
• in the detection of IgG levels in patients with rheumatoid arthritis
• in small bowel biopsies
• in patients with acute myocardial infarction by immunoblotting

Physical form

The solution in 0.01 M phosphate-buffered saline, pH 7.4, containing 15 mM sodium azide

Storage and stability

For continuous use, store at 2-8 ° C for up to one month. For long-term storage, the solution can be frozen in working aliquots. Repeated freezing and thawing, or storing in “frost-free” freezers is not recommended. If slight cloudiness occurs after prolonged storage, rinse the solution by centrifugation before use.

Disclaimer

Unless otherwise stated in our catalog or other company documentation accompanying the product (s), our products are designed for research use only and are not to be used for any other purpose, including but not limited to commercial uses. unauthorized, in vitro diagnostic uses, ex vivo or in vivo therapeutic uses or any type of consumption or application to humans or animals.

Biochemical / Physiological Actions

The IgG antibody subtype is the most abundant of the serum immunoglobulins of the immune system. It is secreted by B cells and is found in blood and extracellular fluids and provides protection against infections caused by bacteria, fungi, and viruses. Maternal IgG is transferred to the fetus through the placenta, which is vital for the newborn’s immune defense against infection. Mutations in the Fc region of IgG are implicated in autoimmune diseases such as rheumatoid arthritis. Modified Fc proteins are of therapeutic importance for the treatment of autoimmune diseases.

Immunogen: Recombinant fragment containing amino acids 511-580 of human Glypican 3 (1G12); Full-length recombinant human GPC3 protein (GPC3 / 863)

Location: Cytoplasmic

Marker: Hepatocellular carcinoma marker

Specificity

Glypican-3 (GPC3) is an integral membrane protein that is mutated in Simpson-Golabi-Behmel syndrome (GBSS). SGBS is characterized by pre-and postnatal overgrowth and is an X-linked recessive condition. GPC3 can also be found in secreted form. Anti-GPC3 has been identified as a useful tumor marker for the diagnosis of hepatocellular carcinoma (HCC), hepatoblastoma, melanoma, testicular germ cell tumors, and Wilms tumor.

In HCC patients, GPC3 is overexpressed in neoplastic liver tissue and elevated in serum, but is undetectable in normal liver, benign liver, and serum from healthy donors. GPC3 expression is also found to be higher in HCC liver tissue than in cirrhotic liver or liver with focal lesions such as dysplastic nodules and areas of liver adenoma (HA) with malignant transformation. In the context of testicular germ cell tumors, GPC3 expression is up-regulated in certain histological subtypes, specifically yolk sac tumors and choriocarcinoma.

A high level of GPC3 expression has also been found in some types of embryonal tumors, such as Wilm’s tumor and hepatoblastoma, with low or undetectable expression in adjacent normal tissue. In thyroid cancer patients, GPC3 expression is dramatically improved in certain types of cancers: 100% in follicular carcinoma and 70% in papillary carcinoma. The expression of GPC3 in follicular carcinoma was significantly higher than that of follicular adenoma. In contrast, GPC 3 is not expressed in anaplastic carcinoma.

Isotype: IgG

Clonality: Monoclonal

Host: Mouse

Gene: GPC3

Purity: Purified Protein A or G

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Applications / Dilutions

Dilutions

• Flow cytometry 0.5 – 1 ug / million cells in 0.1 ml
• Immunocytochemistry / Immunofluorescence 1-2 ug / ml
• Immunohistochemistry
• Immunohistochemistry-Paraffin 0.5 – 1.0 ug / ml
• Immunofluorescence 0.5 – 1.0 ug / ml

Application notes

• Immunohistochemistry (formalin-fixed): 1-2 ug/ml for 30 minutes at RT. Staining of formalin-fixed tissues requires heating tissue sections in 1 mM EDTA buffer, pH 7.5-8.5, for 45 min at 95 ° C followed by cooling at RT for 20 min.
• The optimal dilution for a specific application must be determined.

Theoretical MW

• 67 kDa.
• Disclaimer Note: The observed molecular weight of the protein may vary from the listed predicted molecular weight due to post-translational modifications, post-translation cleavages, relative charges, and other experimental factors.

Packaging, storage, and formulations

Storage: Store at 4C.

Buffer: 10 mM PBS with 0.05% BSA

Preservative: 0.05% sodium azide

Concentration: 0.2mg / ml

Purity: Purified Protein A or G

Alternative names for the Glypican 3 antibody (1G12 + GPC3 / 863)

• DGSX
• Glypican 3
• glypican 3 proteoglycan
• glypican-3
• GPC3
• GTR2-2
• proteoglycan heparan sulfate
• OCI-5 intestinal protein
• MXR7
• OCI5
• OCI-5
• glypican-3 secreted
• SGB
• SGBS
• SGBS1SDYS

Limitations

This product is for research use only and is not approved for human use or clinical diagnosis. Primary Antibodies are guaranteed for 1 year from the date of receipt.

Resume

Anti-glycan antibodies are an abundant subpopulation of serum antibodies with critical functions in many immune processes. Changes in the levels of these antibodies can occur with the onset of disease, exposure to pathogens, or vaccination. As a result, there has been significant interest in exploiting anti-glycan antibodies as biomarkers for many diseases. The serum contains a mixture of anti-glycan antibodies that can recognize the same antigen, and competition for binding can potentially influence the detection of antibody subpopulations that are more relevant to disease processes.

The most abundant antibody isotypes in serum are IgG, IgM, and IgA, but little is known about how these different isotypes compete for the same glucan antigen. In this study, we developed a multiplexed glucan microarray assay and applied it to assess how different isotypes of anti-glucan antibodies (IgA, IgG, and IgM) compete for the imprinted glycan antigens. While IgG and IgA antibodies generally outperform IgM for peptide or protein antigens, we found that IgM outperformed IgG and IgA on many glucan antigens.

To illustrate the importance of this effect, we provide evidence that IgM competition may explain the unexpected observation that IgG of certain antigenic specificities appears to be preferentially transported from mothers to fetuses. We show that IgM in maternal serum competes with IgG resulting in lower than expected IgG signals. Since cord blood contains very low levels of IgM, competition only affects maternal IgG signals, making it appear that certain IgG antibodies are higher in cord blood than matched maternal blood. Taken together, the results highlight the importance of competition for studies involving anti-glycan antibodies.

Introduction

Human serum contains a wide variety of carbohydrate-binding antibodies that play a critical role in human health and provide a rich pool of potential biomarkers for many biomedical applications and diseases. For example, the detection of anti-glycan antibodies against blood group A and B antigens provides a simple and reliable strategy to predict which individuals are suitable for transfusion and transplantation. Anti-glycan antibodies are also crucial in other areas of immunology, such as tumor surveillance, autoimmunity, defense against pathogens, and response to vaccines.

These broader immune functions have stimulated interest in exploring the potential use of circulating anti-glycan antibodies as biomarkers for a wide variety of diseases. Detection of antiglycan antibodies in serum is typically carried out by immobilizing a carbohydrate of interest, capturing specific antibodies, and then measuring the amounts of bound antibodies.

This process is complicated by the fact that serum often contains a mixture of antibodies that recognize the same antigen, and certain antibodies within the mixture may be more relevant to immune protection or disease processes than others. Antibodies against a particular glycan can vary in terms of affinity, specificity, concentration, and/or isotype, but they can all compete to bind to the same antigen. As a result, the binding of one can influence the detection of the others, and it can be difficult to reliably measure a subpopulation of target antibodies of interest.

Results

Our approach to assessing competence involved the use of purified IgG, IgA, and IgM antibodies from pooled human serum. Each polyclonal antibody sample would be profiled on our glycan microarray individually and in the presence of other isotypes. In addition, changes in IgG and IgM anti-glycan antibody signals would be assessed in whole serum after the addition of IgG, IgA, and IgM. Although IgD and IgE are also present in serum at low concentrations and capable of competing, this study focused on the most abundant antibodies in serum.