fluorescent-conjugates

Iizuka K, Yokomizo T, Watanabe N, Tanaka Y, Osato M, Takaku T, Komatsu N (2016) Lack of phenotypical and morphological evidences of endothelial to hematopoietic transition in the murine embryonic head during hematopoietic stem cell emergence. PLoS One 11:e0156427. doi: 10.1371/journal.pone.0156427 PMID: 27227884

Summary: Hemogenic endothelial cells have been observed in several embryonic tissues, such as the dorsal aorta, the placenta and the yolk sac. Recent work also suggest that the mouse embryonic head also produces hematopoietic stem cells (HSCs)/progenitors. However, a histological basis for HSC generation in the head hasn’t been determined because the hematopoietic cluster and hemogenic endothelium have not been well characterized. The authors in this study used whole-mount immunostaining and 3D confocal reconstruction techniques to analyze both c-Kit hematopoietic cluster and Runx1 hemogenic endothelium in the whole-head vasculature. Alexa488 labeled anti-NGFr (Cat. #FL-N01AP) was used in flow cytometry. The number of c-Kit hematopoietic cells was 20-fold less in the head arteries than in the dorsal aorta. In addition, nascent hematopoietic cells, observed by a budding structure and a Runx1 hemogenic endothelium, were not observed in the head. These results suggest that head HSCs may not be or are rarely generated from the endothelium in the same manner as aortic HSCs.

Related Products: NGFr (mu p75) Rabbit Polyclonal, affinity-purified Alexa488-labeled (Cat. #AB-N01APFLA)

Ovsepian SV, Antyborzec I, O’Leary VB, Zaborszky L, Herms J, Oliver Dolly J (2013) Neurotrophin receptor p75 mediates the uptake of the amyloid beta (Abeta) peptide, guiding it to lysosomes for degradation in basal forebrain cholinergic neurons. Brain Struct Funct 219(5):1527-1541. doi: 10.1007/s00429-013-0583-x PMID: 23716278

Summary: Accumulation of β-amyloid in the brain is considered one of the main causes of Alzheimer’s disease. The increase in β-amyloid is accompanied by a reduction in levels of the high affinity nerve growth factor receptor (trkA) and cognitive impairment. The authors looked at levels of the low affinity nerve growth factor receptor (p75) that do not decline. Using a 0.8-μg injection of 192-Cy3 (Cat. #FL-01) into the medial prefrontal cortex of rats the authors assessed the transport of p75 and β-amyloid by microscopy. The results indicate that the primary destinations of both p75 and β-amyloid were the late endosome and lysosome.

Related Products: 192-IgG Mouse Monoclonal, Cy3-labeled (Cat. #AB-N43FL3)

Li Q, Syrovets T, Simmet T, Ding J, Xu J, Chen W, Zhu D, Gao P (2013) Plasmin induces intercellular adhesion molecule 1 expression in human endothelial cells via nuclear factor-κB/mitogen-activated protein kinases-dependent pathways. Exp Biol Med (Maywood) 238(2):176-186. doi: 10.1177/1535370212473700

Summary: Intracellular adhesion molecule 1 (ICAM-1) mediates inflammatory cell migration – an early step in atherosclerosis. The authors investigated an inflammatory cascade activated by plasmin using a variety of methods, including flow cytometry with anti-mouse IgG-FITC (Cat. #FL-07) and anti-rabbit IgG-FITC (Cat. #FL-04).

Related Products: Goat Anti-Rabbit IgG, FITC-labeled (Cat. #FL-04), Goat Anti-Mouse IgG, FITC-labeled (Cat. #FL-07)

Ovsepian SV, Dolly JO, Zaborszky L (2012) Intrinsic voltage dynamics govern the diversity of spontaneous firing profiles in basal forebrain noncholinergic neurons. J Neurophysiol 108(2):406-418. doi: 10.1152/jn.00642.2011 PMID: 22496531

Summary: The voltage modulation functions of the basal forebrain have commonly been associated with cholinergic cells. More recent work has suggested that noncholinergic cells have an influence on this type of neuronal activity. The authors labeled cholinergic neurons by injecting 0.8-1.6 μg of Cy3-192-IgG (Cat. #FL-01) into the lateral ventricles of rats. Patch-clamp recordings were taken from brain slices of these animals under various blockade conditions. The results demonstrate that neuropeptide Y receptors as well as ions such as Ca2+ and K+ are important for regenerative firing in basal forebrain cholinergic neurons.

Related Products: 192-IgG Mouse Monoclonal, Cy3-labeled (Cat. #AB-N43FL3)

Hawryluk JM, Ferrari LL, Keating SA, Arrigoni E (2012) Adenosine inhibits glutamatergic input to basal forebrain cholinergic neurons. J Neurophysiol 107(10):2769-2781. doi: 10.1152/jn.00528.2011 PMID: 22357797

Summary: Using patch-clamp recordings from mouse brain slices the authors demonstrate that adenosine not only directly inhibits cholinergic neurons in the basal forebrain, it also reduces excitatory inputs to these neurons as well. Cy3-anti-mu p75 (Cat. #FL-05, 40-60 ng) was injected into the lateral cerebroventricle.

Related Products: NGFr (mu p75) Rabbit Polyclonal, affinity-purified Cy3-labeled (Cat. #AB-N01APFL3)

Q: I plan to use your Secondary Antibody Conjugates, Rab-ZAP (Cat. #IT-05), and Fab-ZAP Rabbit (Cat. #IT-57) with my primary antibody and would like to observe eliminated cells using a fluorescence microscope. The idea is to co-culture cancer cells and fibroblast cells, and kill fibroblast cells only with a specific primary antibody. Then I want to observe the eliminated fibroblast cells and take pictures with a fluorescence microscope. Can you recommend a protocol?

A: In order to stain and visualize the cells that are being eliminated, it would be best to stain for Saporin using a fluorescently-tagged antibody such as the FITC-labeled Saporin antibody (Cat. #AB-15AP-FL). By washing off the media after a day, and then staining for saporin, one would illuminate only the cells that have internalized the saporin (marking them for death). The cells that do not stain for saporin will live.

Related Product: FITC-labeled Saporin antibody (Cat. #AB-15AP-FL)

Hess PR, Buntzman AS, Murray SL, Young EF, Frelinger JA (2009) Featured Article: Selective deletion of CD8+ T cells by saporin-coupled MHC class I tetramers. Targeting Trends 10(1)

Related Products: Streptavidin-ZAP (Cat. #IT-27), Saporin Goat Polyclonal, affinity-purified FITC-labeled (Cat. #AB-15APFL)

Read the featured article in Targeting Trends.

See Also:

• Hess PR et al. Selective deletion of antigen-specific CD8+ T cells by MHC class I tetramers coupled to the type I ribosome-inactivating protein saporin. Blood 109:3300-3307, 2007.

Yu S, Xie H, Datta A, Naidu N, Gu XX (2008) Galactose residues on the lipooligosaccharide (LOS) of moraxella catarrhalis 26404 (serotype c) form the epitope recognized by the bactericidal antiserum from conjugate vaccination. Infect Immun 76(9):4251-4258. doi: 10.1128/IAI.01570-07

Related Products: Goat Anti-Rabbit IgG, FITC-labeled (Cat. #FL-04)

Huh CY, Danik M, Manseau F, Trudeau LE, Williams S (2008) Chronic exposure to nerve growth factor increases acetylcholine and glutamate release from cholinergic neurons of the rat medial septum and diagonal band of Broca via mechanisms mediated by p75NTR. J Neurosci 28(6):1404-1409. doi: 10.1523/JNEUROSCI.4851-07.2008 PMID: 18256260

Related Products: 192-IgG Mouse Monoclonal, Cy3-labeled (Cat. #AB-N43FL3)

Chin JH, Ma L, MacTavish D, Jhamandas JH (2007) Amyloid beta protein modulates glutamate-mediated neurotransmission in the rat basal forebrain: involvement of presynaptic neuronal nicotinic acetylcholine and metabotropic glutamate receptors. J Neurosci 27:9262-9269. doi: 10.1523/JNEUROSCI.1843-07.2007 PMID: 17728440

Summary: This work focused on the effect of amyloid beta on glutamate-mediated neurotransmission in the diagonal band of Broca. Using neurons identified by staining with Cy3-labeled 192-IgG (Cat. #FL-01, 5 µl of 1:1 diluted antibody injected into the left and right ventricle) the authors monitored the response to amyloid beta by measuring excitatory postsynaptic currents via whole-cell patch-clamp recordings. The results suggest that glutamate neurotransmission might be vulnerable to Alzheimer’s disease, and may also be a therapeutic target.

Related Products: 192-IgG Mouse Monoclonal, Cy3-labeled (Cat. #AB-N43FL3)