Antibody News

What are the major differences between Apoptosis, Necroptosis & Autophagy?

Friday, March 24, 2017 - 13:41

Apoptosis is a form of programmed cell death which is mediated by cysteine proteases called caspases. It is an essential phenomenon in the maintenance of homeostasis and growth of tissues, and it also plays a critical role in immune response. The cytomorphological alterations and the key features of apoptosis are listed below:

apoptosis

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Process

Active, physiological or pathophysiological

Induction stimuli

Oxidative stress, death receptor ligands, chemotherapy

Ultrasensitive IHC Detection with HRP-Polymer Conjugates

Thursday, March 23, 2017 - 14:54

Signal amplification methods are widely used in immunohistochemistry (IHC) for detection of rare epitopes and low abundance antigens. While many of these techniques such as the avidin-biotin complex (ABC) method improve staining, they frequently require additional steps and result in higher background staining.  Blocking endogenous biotin, a requirement of using ABC reagents, may not sufficiently remove residual activity in frozen tissue sections and tissues high in biotin including the liver and kidney.

FABP1/L-FABP antibody

Figure 1. FABP1/L-FABP was detected in paraffin-embedded sections of human kidney using Mouse Anti-Human FABP1/L-FABP Monoclonal Antibody (Catalog # MAB2964) at 1 μg/mL for 1 hour at room temperature followed by incubation for 30 minutes at room temperature with Anti-Mouse/Rabbit IgG VisUCyte HRP Polymer Antibody (Catalog...

The use of apoptosis antibodies and controls in cell death research

Monday, March 20, 2017 - 15:06

Apoptosis is a method of programmed cell death that is notably characterized by a morphological change in cellular nuclei and membrane appearance.  Not to be confused with necrosis, apoptosis is a pathway that is induced by a variety of factors that activate cysteine proteases known as caspases to lead the cell to its ultimate death versus natural death of a cell. While excessive apoptosis can be detrimental, it can also be part of our immune response or in protection of cells that have been damaged by disease or toxic stimuli.  Using antibodies against apoptotic pathway proteins is an effective way to investigate the role of apoptosis in a number of experimental models.  However, it is important to use antibodies against well-defined key apoptotic players and to integrate controls when possible. 

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Frontiers in immunohistochemical (IHC) analysis

Thursday, March 16, 2017 - 09:06

There are a variety of experimental methods to choose from when using antibodies as a probe to highlight a target of interest. Western blot will reveal protein abundance and behavior, immunocytochemistry allows a look at protein behavior on the cellular level, and flow cytometry has the ability to label and sort hundreds of thousands of cells in no time. However, immunohistochemistry (IHC) is the only method where researchers have the ability to view the spatial localization of a target within a specific tissue. A lot has changed with IHC since its introduction in the mid 1900s, including automated processing machines, multiplexing techniques, advanced image analysis, robustly

Epidermal growth factor receptor (EGFR) in head and neck squamous cell carcinoma (HNSCC)

Tuesday, March 14, 2017 - 09:24

Head and neck squamous cell carcinoma is quite common in the U.S., covering more than 4% of all cancers each year, and is most susceptible to individuals between 50 and 60 years of age.  Squamous cells are a type of epithelial cell that are located all over the body with concentrations in the mouth, throat, neck and cervix.  EGFR, or epidermal growth factor receptor, is a trans-membrane glycoprotein that oversees cellular proliferation through its intrinsic tyrosine kinase activity.  When EGFR is bound to its ligand, it is phosphorylated by inner tyrosine kinase activity, where downstream pathways are activated and tumors caused by over stimulated cellular proliferation may occur.  Because of EGFR’s role in regulating multiple signaling cascades, and for the potential treatment therapies targeted at kinase inhibitor activity, EGFR is of high interest in understanding head and neck squamous cell carcinoma.  The following articles used an...

Topics in CD11b: The innate immune response

Thursday, March 9, 2017 - 11:58

Integrins are transmembrane receptors composed of alpha and beta chains, where beta-integrins are mainly expressed in leukocytes. Leukocytes are white blood cells that act in the immune system to defend our body against foreign pathogens.  Integrins are known to interact with extracellular matrix molecules to initiate our inflammatory immune response, in addition to regulating cell adhesion, migration and proliferation.  Our innate immune response is composed of a number of cell types that work in a coordinated effort to identify and attack foreign particles through antigen production.  CD11b is expressed in macrophages, monocytes, dendritic cells and granulocytes – the main players of our immune response cell network.  While CD11b’s exact role in immunity is still debated, it has been established that CD11b both negative and positively regulates our immune response, often times via a TLR ligand.  The following research articles use a...

The role of Parkin and autophagy in retinal pigment epithelial cell (RPE) degradation

Tuesday, March 7, 2017 - 13:49

The root of Parkinson’s disease (PD) points to a poorly regulated electron transport chain leading to mitochondrial damage, where many proteins need to work cohesively to ensure proper function.  The two key players of this pathway are PINK1, also known as PTEN or PARK6, and Parkin, also known as PARK2 - where PINK1 acts as an upstream effector of Parkin to regulate mitochondrial dynamics.  Mitochondria must maintain a healthy equilibrium and do so by undergoing a series of fission and fusion events.  The proteins Drp-1 and OPA-1, respectively, govern these events. While PINK1 and Parkin are directly involved in the progression of PD, their role in maintaining mitochondrial health has associated them with autophagy and mitophagy events in other models such as the loss of retinal pigment epithelial cell (RPE)...

Article Review: Ly6E/K Signaling to TGF-beta Promotes Breast Cancer Progression, Immune Escape, and Drug Resistance

Friday, March 3, 2017 - 14:52

Today in breast cancer research, scientists are focused on determining the cause, risk, diagnostic testing options and treatment of this devastating disease.  Of particular interest is identification of potential therapeutic targets that are known to contribute to the progression of breast cancer in order to develop treatments against these specific genes or proteins.  This article review summarizes research completed by AlHossiny et al regarding the role of Ly6E/K signaling and TGF-beta is the progression of cancer, immune escape and subsequent drug resistance.  Initial work by this group shed light on the role of mouse Ly6A in the regulation of TGF-beta, PTEN and the ERK/AKT signaling pathways to develop resistance to radiation and promote metastatic behavior of mammary tumors.  Furthermore, heightened levels...

The use of the autophagy marker LC3 in western blot, immunocytochemistry and flow cytometry research applications as an indicator of autophagy

Thursday, March 2, 2017 - 08:44

The process of autophagy, or lysosome-mediated degradation of damaged proteins and organelles in the cytosol, is a vital cellular process that acts as a quality control mechanism for proteins and organelles. The misregulation of autophagy can lead to an imbalance of cellular homeostasis and the subsequent development of disease.  Therefore, the study of autophagy is at the forefront of neuroscience and cancer research, among others.  In order to measure autophagic flux, many assays use the autophagy marker protein LC3.  LC3, the mammalian homolog of yeast ATG8, is a ubiquitin like protein that is associated with the autophagosome during the autophagic process. More directly related to LC3 is the process of selective autophagy, where receptors such as p62, NBR1 and NDP52 possess an LC3-interacting region (LIR) to directly bind LC3. Using a

4EBP1 and skeletal muscle protein synthesis

Wednesday, February 22, 2017 - 15:58

Eukaryotic translation initiation factor 4E binding protein 1, or 4EBP1, is an mRNA translational repressor protein that negatively regulates eukaryotic translation initiation factor 4E, or EIF4E.  EIF4E is a protein that forms a complex necessary to block the 5’ ends of mRNA with a 7-methyl-guanosine five-prime cap structure, which is important for normal translation of mRNA.  Specifically, the EIF4E complex recruits 40s ribosome subunits to scan mRNA in order to regulate protein synthesis.  When EIF4E is bound to 4EBP1, it is held in an inactive state, however phosphorylation of 4EBP1 will cause its release.  While 4EBP1 has broad implications in translational research, the following articles hone in on how a 4EBP1 antibody plays a role in the regulation of skeletal muscle protein synthesis.

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The use of Beta Actin (AC-15) as a loading control across multiple species

Friday, February 17, 2017 - 14:46

Actin is a fundamental component of the cytoskeleton, where it has the ability to create and break down actin filament formation in response to various cell needs.  Actin has six highly conserved isoforms, however beta and gamma actin are the two isoforms that are highly and ubiquitously expressed in the cell.  For this reason, measuring beta-actin levels has served as a useful control in research experiments in order to have a baseline of protein expression to compare cell manipulations to.  However, beta actin has other implications in scientific research aside from acting as a housekeeping gene, and plays a role in cell motility, development, wound healing, cell division and more.  The following articles used the Beta Actin (AC-15) antibody as a loading control in chicken, human and rat samples in order to illustrate its conservation and reliability when used across species. 

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The role of HIF-2 alpha in the progression and therapy of clear cell renal cell carcinoma

Monday, February 13, 2017 - 14:51

HIF-2 alpha, also known as hypoxia-inducible factor 2, endothelial PAS domain protein-1, and member of PAS superfamily 2 is part of the HIF family of proteins.  The HIF family is composed of HIF-1, HIF-2 and HIF-3, where HIF-2 is a dimeric protein that consists of an alpha and beta subunit.HIF-2 alphais activated in the presence of oxygen deprivation, or hypoxia, via prolyl hydroxylase-domain enzymes (PHDs) that facilitate its degradation.  HIF-2 alphais expressed in endothelial and intestinal cells, as well as in tumor tissue of a variety of prominent cancers, and is not typically detected in normoxic conditions.  Given the fact that loss of HIF-1 alpha was a notable hallmark of renal cell carcinoma, HIF-2 alpha and the hypoxia mediated gene therapy has been an area of interest in...

Dual applications of a c-Myc antibody in mitochondrial research

Thursday, February 9, 2017 - 08:10

c-Myc, a proto-oncogene, has documented involvement in cellular differentiation, cell growth, cell death and tumor formation.  Target genes of the Myc family include those that participate in cell survival, translation, transcription, metabolism and more.  On a more specific level, c-Myc is a transcription factor that can both activate and repress its target protein by way of DNA modifications.  This allows for the use of a c-Myc antibody in two manners; it can be used to monitor the actual c-Myc protein expression levels, or, it can be used as a probe against a c-Myc protein tagged to another protein of interest. These interactions can be exploited when trying to detect proteins that c-myc interacts with if antibodies against the protein are not available. The structure of c-Myc is what allows for its duality, where its basic-helix-loop-helix-leucine zipper c-term domain can mediate DNA specific binding.  The...

CiteAb 2017 Antibody Awards: The Lifetime Achievement Award Recipient Karen Padgett

Monday, January 30, 2017 - 12:53

The CiteAb Lifetime Achievement Award recognizes and promotes an individual who has made an outstanding contribution to the research antibody industry over a sustained period of time.

We're delighted to announce that Karen Padgett, founder of Novus Biologicals, has been recognized for her lifetime achievement in the research antibody industry. Karen is currently the VP of the Antibody Business Unit (representing both Novus and R&D Systems) and Digital Marketing at Bio-Techne. Karen has earned her place as a global leader in the research antibody industry, through her unfailing commitment, drive and positivity over the last two decades, and without ever compromising the integrity that defines her company as an industry leader.

Citeab award

Over 20 years of...

IHC vs ICC vs IF – Do you know the difference?

Monday, January 30, 2017 - 08:22

Immunohistochemistry (IHC), Immunocytochemistry (ICC) and Immunofluorescence (IF) all utilize antibodies to provide visual details about protein abundance, distribution and localization. These terms are often confusing and are sometimes mistakenly used interchangeably.  Thus, it is important to understand the fundamental differences between these various techniques. 


IHC-Fr (on left): Cadherin‑17 was detected in perfusion fixed frozen sections of mouse intestine using rabbit anti-mouse Cadherin‑17 monoclonal antibody (Catalog # MAB8524...

The importance of using controls in immunohistochemistry

Monday, January 23, 2017 - 08:52

Immunohistochemistry (IHC) is a widely applied experimental method used to examine tissue antigen expression and behavior with the use of an antibody conjugated to a secondary tag for visualization.  IHC consists of a tissue preparation phase, an antibody-staining phase, and a result analysis phase - all of which may lead to skewed results if not properly performed.  One way to ensure that IHC staining results are in fact demonstrating protein behavior and not a side effect the experimental process is to use IHC controls within each experiment.  By doing so, you can feel confident in your protein integrity, experimental methods and primary and secondary antibody efficacy. 

To begin, the manner in which tissue is harvested, dissected and mounted to a microscope slide is very important.  It is vital that the tissue is collected rapidly in order for the proteins to...

The role of p53 in UV radiation DNA damage and subsequent tumorogenesis

Friday, January 20, 2017 - 13:19

p53, the protein product of the tp53 gene, is one of the most widely studied tumor suppressor proteins in cancer research.  p53 is unique in that it demonstrates both tumor suppressive and tumor progressive properties depending on whether it is functional or mutated.  The most common mutation in the p53 protein that leads to lack of tumor suppression activity is a missense mutation, however frameshift or nonsense mutations are also common.  In fact, mutant p53 has exhibited dominant negative inhibition of the wild type version of the protein, demonstrating the fact that the p53 pathway has both positive and negative feedback loops.  The p53 pathway is a large signal transduction pathway that affects DNA replication and division, apoptosis, cellular senescence and is composed of at least seven negative feedback loops and three positive feedback loops.  Although p53's major influence is in cancer, the following articles describe how a...

The application of CD31/Pecam-1 (MEC 7.46) in breast cancer research

Monday, January 16, 2017 - 15:27

CD31/PECAM-1, or platelet endothelial cell adhesion molecule 1, is a 130-kDa glycoprotein expressed on vascular and hematopoietic cells.  Depending on the cell type, CD31/PECAM-1 expression can be largely localized to cell junctions, playing a role in cell adhesion.  Aside from its role in cellular adhesion, CD31/PECAM-1 is also a large player in a variety of signaling pathways, such as angiogenesis, cell migration, leukocyte transmigration and more.  Specifically, the association and indication of angiogenesis in breast cancer is of interest, given that extensive research shows angiogenesis is involved in breast cancer development, invasion and metastasis.  The use of a CD31/PECAM-1 primary antibody in experimental studies to quantify tumor neovascularization is a popular approach to measure microvessel density.

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The use of actin as a loading control in research on fruiting-body development and vegetative growth in Sordaria macrospora research

Monday, January 9, 2017 - 14:46

Sordaria macrospora is a filamentous fungus that serves as very useful system for scientific research due to a short life cycle and easy manipulation.  Just like any other model organism, it is important to have an effective loading control to validate experiments in the Sordaria macrospora.  In addition, the growth and morphogenesis of filamentous fungi is dependent on actin organization.  Actin is a very abundant protein across biological species and can transition between monomeric and filamentous states.  Actin has an alpha and beta isoform, both of which share nearly 98% percent sequence homology with total actin.  Actin recruits a variety of actin regulators in order to carry out many cellular functions, including cell division, cell motility, cell growth and differentiation, muscle contraction and more.  The following articles use an actin antibody as a loading control in the study of...

The C99 fragment of amyloid precursor protein (APP)

Friday, January 6, 2017 - 08:11

Alzheimer’s Disease (AD) is a neurodegenerative disorder that is characterized by an abundance of the beta-amyloid peptide in the brain.  When AD was first discovered, it was determined that beta-amyloid was produced as a result of the proteolysis of the amyloid precursor protein (APP).  Aside from its role in AD, the single-pass transmembrane APP has a high expression level in the brain and tends to concentrate at the synapses of neurons.  Because of this localization, it has been suggested that APP plays a role in synapse formation and potentially plasticity.  However, the exact function of APP is not known.  While there is currently no answer as to why APP accumulates in the brain of elderly individuals, current therapeutic approaches tend to focus on treatments that affect APP processing.  Over the past few years, the structure of APP has been closely studied.  From this research came the discovery that amyloid-beta production requires two cleavage steps...

Pyruvate Dehydrogenase E1-alpha subunit: Understanding the Catalysts in Glycolysis

Thursday, December 29, 2016 - 14:01

Pyruvate Dehydrogenase E1-alpha subunit (PDHA1) is one of multiple enzymes in a mitochondrial complex, called the Pyruvate Dehydrogenase (PDH) complex, involved in the transition between glycolysis and the tricarboxylic acid (TCA) cycle during cellular respiration. The PDHA1 enzyme catalyzes the reaction that produces acetyl-CoA and CO2 from pyruvate (1). PDHA1 also regulates the PDH complex through reversible phosphorylation.

PDHA1 antibody

Pyruvate Dehydrogenase E1-alpha subunit [p Ser293] Antibody [NB110-93479] - Pyruvate Dehydrogenase E1-alpha subunit [p Ser293] antibody (1:250) was tested in HeLa cells with Dylight 488 (green). Nuclei and alpha-tubulin were counterstained with DAPI (blue) and Dylight 550 (red)....

The dynamic use of a PCNA antibody in fish, porcine and primate species

Tuesday, December 27, 2016 - 14:39

Proliferating cell nuclear antigen (PCNA) plays a crucial role in nucleic acid metabolism as it pertains to DNA replication and repair.  Most noted for its activation of subunits of DNA polymerase, it has also been found to interact with cell-cycle progression proteins.  Modifications of PCNA as a result of cellular response put PCNA in a pivotal position with DNA replication, DNA damage, and chromatin structure and function.  In response to DNA damage, PCNA is ubiquitinated and becomes part of the RAD-6 dependent DNA repair pathway, where it acts as a substrate with a variety of proteins. In DNA replication, PCNA acts as a sliding clamp that localizes proteins to DNA strands.  Additionally, the presence of PCNA at DNA maintenance sites where delayed replication forks are found further points to its role in DNA damage and repair. The PCNA mouse monoclonal antibody (PC10) from Novus Biologicals has been effective to...

The use of a GFP antibody for research applications in transgenic C. elegans, GFP tagged yeast and porcine model

Monday, December 19, 2016 - 15:32

GFP, or green fluorescent protein, is a chemiluminescent protein derived from Aequorea jellyfish that was first discovered by Osamu Shimomura.  It was soon after established that the emission spectra of GFP was right around 509nm, or the ultraviolet color range.  The GFP gene is often used to form expression constructs in order to closely follow protein behavior, cellular differentiation, protein localization and more.  The following articles employed a GFP antibody in conjunction with various other GFP construct techniques to strengthen their research findings. 

GFP antibody

Immunocytochemistry/Immunofluorescence: GFP Antibody [NB600-308] - Analysis of GFP in transgenic mouse pancreas in OCT. Verified customer review from...

The Key Benefits of Indirect Detection

Tuesday, December 13, 2016 - 11:50

Even though the direct detection method is becoming more popular for immunofluorescence (IF) and flow cytometry experiments, the indirect detection method still remains the preferred choice for many other applications. In direct detection, the labeled primary antibody is responsible for both binding and detection of the antigen of interest. In indirect detection, this process is broken down into at least two distinct steps – (i) an unconjugated primary antibody forms a complex with the antigen, (ii) a labeled secondary antibody, interacting with the constant region of the primary antibody, facilitates detection.

HAl-1 antibody

HAI-1 was detected in paraffin-embedded sections of human lung cancer using goat anti-human HAI-1 ectodomain antigen affinity-purified polyclonal antibody (Catalog # ...

Top 4 reasons: why use CRISPR-Cas9 antibodies and how?

Friday, December 9, 2016 - 14:37
1. Verification of the success of transfection

Why- If the CRISPR-Cas9 transfection is not successful, it would not be relevant to relate the observations from transfected cells to the expected outcome of gene editing experiment.

How- CRISPR-Cas9’s successful transfection can be verified through its detection at the protein level by employing Western blot or confocal staining analysis of cells that were subjected to CRISPR-Cas9 transfection.

2. Localization of CRISPR-Cas9 at subcellular level

Why- CRISPR-Cas9 must translocate to the nuclei of the transfected cells for executing its nuclease activity on the genomic DNA. Therefore, one must check that the Cas9 protein is actually being delivered into the nucleus.

How - The...

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