Tools

To characterize the iPSC differentiation into pancreatic progenitors we are using the following antibodies:

Mouse monoclonal antibody [PC10] to PCNA (ab29)

Proliferating cell nuclear antigen (PCNA) is an auxiliary protein of DNA polymerase delta and is involved in the control of eukaryotic DNA replication by increasing the polymerase’s processibility during elongation of the leading strand. PCNA is a homotrimer and achieves its processivity by encircling the DNA, where it acts as a scaffold to recruit proteins involved in DNA replication, DNA repair, chromatin remodeling and epigenetics.

Rabbit monoclonal antibody [UMB2] to CXCR4 (ab124824)

C-X-C chemokine receptor type 4 (CXCR-4) also known as fusin or CD184 (cluster of differentiation 184) is an alpha-chemokine receptor specific for stromal-derived-factor-1 (SDF-1 also called CXCL12), that transduces a signal by increasing intracellular calcium ions levels and enhancing MAPK1/MAPK3 activation. Acts as a receptor for extracellular ubiquitin; leading to enhance intracellular calcium ions and reduce cellular cAMP levels.

Rabbit monoclonal antibody [EPR14638] to – N-terminal Nkx2.2 (ab191077)

Nkx2.2 is a homeodomain transcription factor that is essential for the differentiation of islet α- and β-cells. Nkx2.2 is expressed early in pancreatic progenitors beginning at embryonic day 9.5 and is ultimately restricted to β-cells and a subset of α-cells and PP cells. In Nkx2.2 null mice, β-cells and the majority of α-cells fail to differentiate and are replaced by ghrelin-producing cells.

Rabbit polyclonal antibody to NeuroD1 (ab16508)

Neurogenic differentiation 1 (NeuroD1), also called β2, is a member of the NeuroD family of basic helix-loop-helix (bHLH) transcription factors. The protein forms heterodimers with other bHLH proteins and activates transcription of genes that contain a specific DNA sequence known as the E-box. It regulates expression of the insulin gene, and mutations in this gene result in type II diabetes mellitus.

Rabbit polyclonal antibody to MAFA (ab98859)

MafA, also known as RIPE3b1,ia a transcription factor that promotes pancreatic development. Also specifically binds the insulin enhancer element RIPE3b and activates insulin gene expression. Cooperates synergistic with NEUROD1 and PDX1.

Mouse monoclonal antibody [3B5] to Neurogenin3 (ab87108)

Sheep polyclonal antibody to Neurogenin3 (AF3444)

Neurogenin 3 (Ngn3) is another member of the bHLH family of transcription factors. Ngn3 is a crucial component in pancreatic development and plays a supporting role in intestinal as well as neuronal cell development. Studies have demonstrated that knockout of Ngn3 in mice leads to death shortly after birth possibly due to after effects of severe diabetes.

Guinea pig polyclonal antibody to PDX1 (ab47308)

Goat polyclonal antibody to PDX1 (AF2419)

Pancreatic and duodenal homeobox 1, also known as insulin promoter factor 1, is a transcription factor necessary for pancreatic development and β-cell maturation. It activates insulin, somatostatin, glucokinase, islet amyloid polypeptide and glucose transporter type 2 gene transcription. Is particularly involved in glucose-dependent regulation of insulin gene transcription. Binds preferentially the DNA motif 5′-[CT]TAAT[TG]-3′. At adult stage, Pdx1 is required for maintaining the hormone-producing phenotype of the pancreatic β-cell.

Rabbit polyclonal antibody to Nkx6.1 (NBP1-82553)

In the pancreas, NKX6.1 is required for the development of beta cells and is a potent bifunctional transcription regulator that binds to AT-rich sequences within the promoter region of target genes.

Rabbit polyclonal antibody to FOXA2 (ab23630)

Forkhead box protein A2 (FOXA2), also known as hepatocyte nuclear factor 3-beta (HNF-3B) or transcription factor 3B (TCF-3B) is a member of the forkhead class of DNA-binding proteins. This transcription factor is involved in embryonic development, establishment of tissue-specific gene expression and regulation of gene expression in differentiated tissues. Is thought to act as a ‘pioneer’ factor opening the compacted chromatin for other proteins through interactions with nucleosomal core histones and thereby replacing linker histones at target enhancer and/or promoter sites. FOXA2 is involved in glucose homeostasis; regulates the expression of genes important for glucose sensing in pancreatic beta-cells and glucose homeostasis. Involved in regulation of fat metabolism.

Mouse monoclonal antibody [RCK108] to CK19 (M0888)

Cytokeratins are proteins of keratin-containing intermediate filaments found in the intracytoplasmic cytoskeleton of epithelial tissue.

Rabbit monoclonal antibody [EP1098Y] to Synaptophysin (NB110-57606)

Synaptophysin, also known as the major synaptic vesicle protein p38, is a  synaptic vesicle glycoprotein with four transmembrane domains weighing 38kDa. It is present in neuroendocrine cells and in virtually all neurons in the brain and spinal cord that participate in synaptic transmission. It acts as a marker for neuroendocrine tumors, and its ubiquity at the synapse has led to the use of synaptophysin immunostaining for quantification of synapses.

Goat polyclonal antibody to Sox9 (AF3075)

Transcription factor SOX-9 recognizes the sequence CCTTGAG along with other members of the HMG-box class DNA-binding proteins. In the pancreas, Sox9 is expressed in all epithelial cells at early developmental stages; its expression is confined to the duct cells as development proceeds. Differentiated acinar and endocrine cells do not express Sox9.

Rabbit polyclonal antibody to C-peptide (ab82696)

The connecting peptide, or C-peptide, is a short 31-amino-acid protein that connects insulin’s alpha-chain to its beta-chain in the proinsulin molecule. C peptide is released into the blood stream in equal amounts to insulin. A test of C peptide levels will show how much insulin the body is making.

Guinea pig polyclonal antibody to Insulin (A0564)

Insulin is a peptide hormone produced by beta cells in the pancreas. It regulates the metabolism of carbohydrates and fats by promoting the absorption of glucose from the blood to skeletal muscles and fat tissue and by causing fat to be stored rather than used for energy. Insulin also inhibits the production of glucose by the liver.

Goat polyclonal antibody to Somatostatin (sc-7819)

Somatostatin, also known as growth hormone-inhibiting hormone (GHIH) or somatotropin release-inhibiting factor (SRIF) is a peptide hormone that regulates the endocrine system and affects neurotransmission and cell proliferation via interaction with G protein-coupled somatostatin receptors and inhibition of the release of numerous secondary hormones. Somatostatin inhibits insulin and glucagon secretion.

Mouse monoclonal antibody to Glucagon [K79bB10]

Glucagon is a peptide hormone, produced by alpha cells of the pancreas, that raises the concentration of glucose in the bloodstream. Its effect is opposite that of insulin, which lowers the glucose concentration. The pancreas releases glucagon when the concentration of glucose in the bloodstream falls too low. Glucagon causes the liver to convert stored glycogen into glucose, which is released into the bloodstream.