Antibody News

Hypoxia-inducible factor (HIF) is a DNA-binding protein that regulates homeostasis via transcription of a wide range of genes. It is inactive in oxygenated cells, but becomes active in hypoxic, i.e. low oxygen conditions. HIF proteins consist of a heterodimeric complex of identical alpha and beta sub-units. The α is degraded in normal oxygen conditions, while the β remains independent of O2 concentration.

HIF is implicated in a wide range of tumours. Recently, we at Novus Biologicals cultured the H1 alpha67 hybridoma cell line to produce the monoclonal anti-HIF 1 alpha antibody. This is one of a number of products in our hypoxia antibody catalogue routinely used in cancer research.

The involvement of HIF-1 in tumour development is obvious when you consider the complexity of the hypoxia pathway. Under normoxic conditions, the α-subunit undergoes prolyl hydroxylation and is tagged by E3 ubiquitin ligase for degradation by the cell...

All cancer genomes carry somatic mutations. These include base substitutions, rearrangements, insertions and deletions (indels). Antibody suppliers have seen tremendous growth in the field of cancer research recently, with antibodies to at least 400 cancer genomes.

Driver mutations are direct somatic changes which confer an advantage for clonal growth and can lead to oncogenesis. Passengers or “silent mutations” do not lead to cancer development, but give important information on pre-cancerous cell changes. Epigenetic mutations do not alter the genetic code, but can still be passed on to at least one successive generation. All are of interest.

DNA damage, such as strand breaks, is common in normal cells, and most do not cause mutation. The changes are recognised by cell enzymes, and various DNA repair mechanisms, such as NER (nucleotide excision repair) correct the damage and prevent driver...

Antibody suppliers have seen an enormous growth in the area of DNA repair in recent years. DNA damage causes structural changes within the cell that can cause genome mutations to occur, a common cause of tumour development and an important area of research for antibody suppliers like us at Novus Biologicals.

NER (nucleotide excision repair) is a global genome repair process, activated by DNA damage caused by (among other things) UV light. This is a common cause of malignant melanomas. NER repairs distortions of the DNA helix by excising the oligonucleotide (short strand section) containing the lesion. Reproductive DNA polymerases fill the resultant gap using the undamaged strand as a template. There is also a transcription-coupled form of NER, which prevents modification of the transcription process by damaged DNA. This appears to be more efficient at preventing mutations, which usually occur at...

Research facilities often require a supply of antibodies of all classes, in both their whole and fragmented forms. There are both pros and cons to using fragments in place of entire proteins. As IgG is the most prevalent class on our antibody database, we’ll cover that one first.

IgG consists of two heavy chains and two light chains, held together by hydrophobic interactions and disulphide bonds. Secondary (anti-IgG) antibodies are derived by injecting an animal of one species (e.g. a sheep) with whole Ig from another species (e.g. mouse) that produced the 1° Ab to the antigen. This results in polyclonal secondary Ab which reacts to both the H and L chains. It reacts to every possible primary epitope, resulting in the largest possible signal.

However, the same light chains are common in all the antibody classes,...

FANCD2 is one of a number of proteins in the FANC group. It undergoes modification in both its normal and disease state; therefore our FANCD2 antibodies are modified and conjugated in various ways for research.

FANCD2 (Fanconi Anaemia Complementation Group D2) is one of several FANC proteins involved with development of Fanconi Anaemia, a genetic disease that increases cell susceptibility to tumour development. In its normal pathway it interacts with several other genes known to cause cancer, for example BRCA1.

FANCD2 is routinely modified by interaction with other proteins, to enable it to carry out normal tasks. The protein occurs in all cells and plays a role in DNA-repair, inhibiting DNA synthesis following exposure to ionizing radiation, as well as resisting cross-linking. It also activates S-phase checkpoints following phosphorylation by ATM. It has been shown FANCD2 may also be part of the DNA repair mechanism, repairing...

Not everyone knows how antibodies influence animal life, so this article is, in part, a very brief introduction. Novus Biologicals offers a printed version of their Apoptosis Catalog that contains additional information on apoptotic mechanisms; email promotions@novusbio.com to get a copy.

Apoptosis is triggered by a variety of signals and takes place by one of several mechanisms. These involve cell surface receptors, cytokines, tumour necrosis factors, proteins such as Bcl-2, Bax and Apaf-1, the caspase cascade, and apoptosis-inducing factors.

Specific antibodies can assist in pinpointing exactly which factors and signals are involved in any particular situation.

Apoptosis is vital in the breakdown of early embryonic structures, so as to permit development and maturation. Without apoptosis, the caterpillar cannot metamorphose into the...

Autism is a condition which demands more research - even if some autistic people do not want their condition to be ‘cured’, most would like some of their symptoms to be alleviated.

Fatemi and colleagues (J Autism Dev Disord. 2009, 39(2) pp. 223–230) subjected brain tissue from autistic and control subjects to SDS-PAGE followed by Western blotting and incubation with antibodies to four GABA-A receptor subunits.

Levels of GABRB3, which was identified using an antibody from the Novus Biologicals antibody catalog, were decreased in the cerebellum and parietal cortex of autistic subjects. The cerebellum also showed decreased levels of GABRA1. The parietal cortex showed decreased levels of GABRA1, GABRA2 and GABRA3. Only GABRA1 was decreased in the frontal cortex of autistic subjects.

Researchers can manipulate antibodies to yield very impressive research outcomes, which may prove highly relevant to the medical treatment of the future.

Severson et al (MBC 2009, 20(7) pp. 1916-1925) used antibodies, including monoclonal rabbit anti-JAM-A and monoclonal rabbit anti-β1 integrin from Novus Biologicals to investigate epithelial cell migration.

In a previous study, this team demonstrated that cell migration is regulated by junctional adhesion molecule A. JAM-A is a transmembrane protein in tight junctions, namely areas where membranes of adjacent cells join closely together. In epithelial cells, JAM-A is thought to decrease paracellular permeability and favour epithelial cell migration. These functions maintain the barrier function of intestinal epithelium. Barrier function is compromised in some intestinal diseases. Stress can cause further dysfunction. The net result is...

Antibodies to beclin 1 may help elucidate the significance of this protein and its role in autophagy.

Autophagy is the lysosomal degradation of cell constituents. It occurs in malnutrition, enabling the body to recycle cell components to provide metabolic precursors. Autophagy is also involved in the response to pathogens and in other processes. Some aspects are summarised here.

Western Blot: Beclin 1 Antibody

Homozygous beclin 1-/- embryos from chimeric beclin 1+/- mice all died in early embryogenesis. This was considered to be due to defective visceral endoderm, in which beclin 1 is normally strongly expressed. Furthermore,...

Anti-AGE antibodies are not the elixir of youth as their name might imply! They have nevertheless provided new information about Alzheimer’s disease, the major cause of dementia in old age.

Advanced glycation end products (AGE) are produced by glycation and oxidation reactions between sugars and free amino groups on proteins and amino acids. They are implicated in a wide range of pathologies. AGE can form in cooked foods. In addition, glycation reactions within the blood form endogenous AGE. High blood sugar levels favour endogenous AGE production.

Diabetes is a risk factor for Alzheimer’s. This motivated a study by Burdo et al (Neurobiol. Aging 2009, 30(12), pp. 1910-1917). The authors used anti-AGE antibodies from Novus Biologicals to detect AGE...

Immunohistochemical testing using antibody panels has a valuable role in cancer diagnosis. Some tumors, especially more malignant ones, tend to lose the appearance of the tissue of origin. However, knowing the tissue of origin assists the physician in selecting the best treatment thus a differential diagnosis is required.

One such example is a tumor containing large cells that somewhat resembles epithelium cells. This could be an undifferentiated melanoma, lymphoma or carcinoma. In order to distinguish between these possibilities, it is recommended to test the tumor tissue using a panel of antibodies against the following substances: keratin, epithelial membrane antigen (EMA), vimentin, leukocyte common antigen (CD45), S100 protein and HMB-45. The pattern of positive versus negative reactions will enable the type of the tumor to be determined.

The vast antibody catalog...

Western blot is a widely used immunoassay technique, used to identify proteins. Many people enter Western blot research without having a clear idea of how the technique relates to antibody usage, so we at Novus Biologicals thought it would be interesting to give a general review of this subject.

Western blotting, also called protein immunoblotting, uses gel electrophoresis to separate proteins of tissue extracts and homogenates. Native and denatured proteins are identified by their isoelectric point, electric charge, molecular weight or molecular structure (often, a combination of these).

Tissue for Western Blot analysis is blended or homogenized to break down cells and release the proteins. Bacterial and viral proteins are also identified using Western blot, and samples are not restricted to cellular tissues...