Immunohistochemical analysis of paraffin-embedded human ductal breast carcinoma using VHL (E3X9K) Rabbit mAb.
Immunohistochemical analysis of paraffin-embedded Renca syngeneic tumor using VHL (E3X9K) Rabbit mAb.
| Cat. # | Size | Qty. | Price |
|---|---|---|---|
| 43065T | 1 Kit (6 x 20 microliters) |
|
| Product Includes | Quantity | Applications | Reactivity | MW(kDa) | Isotype |
|---|---|---|---|---|---|
| HIF-1α (E1V6A) Rabbit mAb 48085 | 20 µl |
|
H M | 120 | Rabbit IgG |
| HIF-1β/ARNT (D28F3) XP® Rabbit mAb 5537 | 20 µl |
|
H M R Mk | 87 | Rabbit IgG |
| VHL (E3X9K) Rabbit mAb 81292 | 20 µl |
|
H M | Rabbit IgG | |
| p300 (D8Z4E) Rabbit mAb 86377 | 20 µl |
|
H Mk | 300 | Rabbit IgG |
| SirT1 (1F3) Mouse mAb 8469 | 20 µl |
|
H M R Mk | 120 | Mouse IgG1 |
| GSK-3β (D5C5Z) XP® Rabbit mAb 12456 | 20 µl |
|
H M R Mk | 46 | Rabbit IgG |
| PKM2 (D78A4) XP® Rabbit mAb 4053 | 20 µl |
|
H M R Mk | 60 | Rabbit IgG |
| LDHA (C4B5) Rabbit mAb 3582 | 20 µl |
|
H Mk | 37 | Rabbit IgG |
| Glut1 (E4S6I) Rabbit mAb 73015 | 20 µl |
|
H M R Mk | 45-60 | Rabbit IgG |
Product Information
Monoclonal antibodies are produced by immunizing animals with synthetic peptides corresponding to residues surrounding Ala475 of human HIF-1α protein, Ile479 of human HIF-1β/ARNT protein, Ser406 of human PKM2 protein, near the carboxy terminus of human p300 protein and human Glut1 protein, and corresponding to the sequence of human LDHA protein.
Monoclonal antibodies are also produced by immunizing animals with recombinant proteins specific to human VHL protein, representing the central region of human SirT1 protein, and specific to the carboxy terminus of human GSK-3β protein.
Hypoxia-inducible factor 1 (HIF1) is a heterodimeric transcription factor that plays a critical role in the cellular response to hypoxia (1). The HIF1 complex consists of two subunits, HIF-1α and HIF-1β, which are basic helix-loop-helix proteins of the PAS (Per, ARNT, Sim) family (2). HIF1 regulates the transcription of a broad range of genes that facilitate responses to the hypoxic environment, including genes regulating angiogenesis, erythropoiesis, cell cycle, metabolism, and apoptosis. The widely expressed HIF-1α is typically degraded rapidly in normoxic cells by the ubiquitin/proteasomal pathway. Under normoxic conditions, HIF-1α is proline hydroxylated leading to a conformational change that promotes binding to the von Hippel-Lindau protein (VHL) E3 ligase complex; ubiquitination and proteasomal degradation follows (3,4). Both hypoxic conditions and chemical hydroxylase inhibitors (such as desferrioxamine and cobalt) inhibit HIF-1α degradation and lead to its stabilization. In addition, HIF-1α can be induced in an oxygen-independent manner by various cytokines through the PI3K-AKT-mTOR pathway (5-7). HIF-1β is also known as AhR nuclear translocator (ARNT) due to its ability to partner with the aryl hydrocarbon receptor (AhR) to form a heterodimeric transcription factor complex (8). Together with AhR, HIF-1β plays an important role in xenobiotics metabolism (8). In addition, a chromosomal translocation leading to a TEL-ARNT fusion protein is associated with acute myeloblastic leukemia (9). Studies also found that ARNT/HIF-1β expression levels decrease significantly in pancreatic islets from patients with type 2 diabetes, suggesting that HIF-1β plays an important role in pancreatic β-cell function (10). CBP (CREB-binding protein) and p300 are highly conserved and functionally related transcriptional co-activators that associate with transcriptional regulators and signaling molecules, integrating multiple signal transduction pathways with the transcriptional machinery (11,12). CBP/p300 also contain histone acetyltransferase (HAT) activity, allowing them to acetylate histones and other proteins (12). The Silent Information Regulator (SIR2) family of genes is a highly conserved group of genes that encode nicotinamide adenine dinucleotide (NAD)-dependent protein deacetylases, also known as class III histone deacetylases. The first discovered and best characterized of these genes is Saccharomyces cerevisiae SIR2, which is involved in silencing of mating type loci, telomere maintenance, DNA damage response, and cell aging (13). SirT1, the mammalian ortholog of Sir2, is a nuclear protein implicated in the regulation of many cellular processes, including apoptosis, cellular senescence, endocrine signaling, glucose homeostasis, aging, and longevity. Targets of SirT1 include acetylated p53 (14,15), p300 (16), Ku70 (17), forkhead (FoxO) transcription factors (17,18), PPARγ (19), and the PPARγ coactivator-1α (PGC-1α) protein (20). Glycogen synthase kinase-3 (GSK-3) was initially identified as an enzyme that regulates glycogen synthesis in response to insulin (21). GSK-3 is a ubiquitously expressed serine/threonine protein kinase that phosphorylates and inactivates glycogen synthase. GSK-3 is a critical downstream element of the PI3K/Akt cell survival pathway whose activity can be inhibited by Akt-mediated phosphorylation at Ser21 of GSK-3α and Ser9 of GSK-3β (22,23). Pyruvate kinase is a glycolytic enzyme that catalyzes the conversion of phosphoenolpyruvate to pyruvate. In mammals, the M2 isoform (PKM2) is expressed during embryonic development (24). Lactate dehydrogenase (LDH) catalyzes the interconversion of pyruvate and NADH to lactate and NAD+. The major form of LDH found in muscle cells is the A (LDHA) isozyme (25). Glucose transporter 1 (Glut1, SLC2A1) is a widely expressed transport protein that transports a number of different aldose sugars into cells (26,27).
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