References

Toledo C, Andrade DC, Del Rio R (2019) Brainstem pre-sympathetic neurons contribute to irregular breathing patterns in volume overload heart failure. FASEB J 33(1):lb630. Experimental Biology 2019 Meeting Abstracts doi: 10.1096/fasebj.2019.33.1_supplement.lb630

Objective: To investigate the contribution of RVLM-C1 neurons on breathing disorders in chronic heart failure (CHF).

Summary: RVLM-C1 neurons play a critical role in the maintenance of altered breathing patterns in CHF rats and highlighted their contribution to the worsening of cardiac function during central chemoreflex activation. DBH-SAP treatment decreased active expiration in CHF rats and deleterious effects of central chemoreflex activation on diastolic cardiac function and cardiac autonomic control were blunted.

Usage: Stereotaxic bilateral injections of Anti-DBH-SAP (5 ng/150 nl).

Related Products: Anti-DBH-SAP (Cat. #IT-03)

Sajjadi E, Seven YB, Simon AK, Zwick A, Satriotomo I, Mitchell GS (2019) Adenosine 2A receptor inhibition promotes neuroprotection following toxic insult to phrenic motor neurons. FASEB J 33(1):844.3. Experimental Biology 2019 Meeting Abstracts doi: 10.1096/fasebj.2019.33.1_supplement.844.3

Objective: The authors explored the role of A2A receptors in phrenic motor neuron cell death in vivo.

Summary: A2A receptors, which contribute to motor neuron death during toxic insults, are upregulated in spared phrenic motor neurons of CTB-SAP treated rats. This is an important finding since A2A receptor upregulation may accelerate motor neuron death in neurodegenerative diseases like ALS.

Usage: CTB-SAP selectively killed nearly all phrenic motor neurons within a week and caused diaphragm paralysis (p<0.01).

Related Products: CTB-SAP (Cat. #IT-14)

Borkowski LF, Nichols NL (2019) A2A and 5‐HT receptors are differentially required for respiratory plasticity over the course of motor neuron loss in intrapleurally CTB-SAP treated rats. FASEB J 33(1):843.3. Experimental Biology 2019 Meeting Abstracts doi: 10.1096/fasebj.2019.33.1_supplement.843.3

Objective: To investigate the role of serotonin (5-HT) and adenosine 2A (A2A) receptors in respiratory plasticity.

Summary: A2A receptors are necessary for respiratory plasticity early (7d), but 5-HT receptors are required late (28d).

Usage: Bilateral, intrapleural injections of: 1) CTB-SAP (25 μg), or 2) un-conjugated CTB and SAP (control) in rats.

Related Products: CTB-SAP (Cat. #IT-14)

Krajewski-Hall SJ, Miranda Dos Santos F, McMullen NT, Blackmore EM, Rance NE (2019) Glutamatergic neurokinin 3 receptor neurons in the median preoptic nucleus modulate heat-defense pathways in female mice. Endocrinology 160(4):803-816. doi: 10.1210/en.2018-00934

Objective: To characterize the thermoregulatory role of MnPO NK3R neurons in female mice.

Summary: Study suggests that KNDy neurons modulate thermosensory pathways for heat defense indirectly via a subpopulation of glutamatergic MnPO neurons that express NK3R.

Usage: Mice were bilaterally injected with 10 ng NK3-SAP in 100 nL PBS (n = 14) or blank-SAP (n = 8) in the preoptic area adjacent to the MnPO.

Related Products: NKB-SAP (Cat. #IT-63), Blank-SAP (Cat. #IT-21)

Shramm PA, Ancheta L (2019) Streptavidin-pHast: A readily conjugatable, pH-sensitive dye to screen for internalization. Cancer Res 79(13 Suppl):2177. Proceedings of the American Association for Cancer Research Annual Meeting, Atlanta GA. PMID: 909090

Summary: Quick and efficient screening of targeting agents that internalize effectively is vital for determining their suitability as potential therapeutics. Some of the most recent successes in the treatment of cancers have been from antibodies to cell surface proteins that are responsible for tumor cell proliferation. Examples are Cetuximab (target: EGFR) approved for colorectal cancer, and Trastuzumab (target: HER2) for breast cancer. These antibodies have more than one effect on the cancer cell, but one of the most important is that, upon binding to the cell surface antigen, the complex is internalized. As such, the down-regulated cell surface protein no longer plays a role in cancer cell division. Here we describe a method for determining internalization of cell surface molecules by targeting agents using a pH-dependent fluorescent reporter cross-linked to streptavidin. Streptavidin is a tetrameric protein (molecular weight 53 kDa in its recombinant form), with each subunit able to bind a single biotin molecule. The bond between streptavidin and biotin is rapid and essentially non-reversible, unaffected by most extremes of pH, organic solvents, and denaturing reagents. It is the strongest known noncovalent biological interaction (Ka = 1015 M-1) between protein and ligand. A variety of molecules, including lectins, proteins, and antibodies, can be biotinylated and reacted with streptavidin-labeled probes or other detection reagents for use in biological assays. The fluorescence from this reporter increases intensity as the pH of its surroundings becomes more acidic, as demonstrated when exposed to the environment inside a cell (thereby providing evidence of internalization). Here we describe methods that can be used to explore candidates as cancer therapeutics in a quick, reliable and reproducible manner.

Related Products: Streptavidin-pHast (Cat. #PH-03)

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See Also:

• Zangardi ML et al. Sacituzumab for the treatment of triple-negative breast cancer: the poster child of future therapy?. Expert Opin Investig Drugs 28(2):107-112, 2019.
• Lamb YN Inotuzumab ozogamicin: First global approval. Drugs 77(14):1603-1610, 2017.
• Kinneer K et al. SLC46A3 as a potential predictive biomarker for antibody-drug conjugates bearing noncleavable linked maytansinoid and pyrrolobenzodiazepine warheads. Clin Cancer Res 24(24):6570-6582, 2018.
• Facciabene A et al. Tumour hypoxia promotes tolerance and angiogenesis via CCL28 and Treg cells. Nature 475:226-230, 2011.
• Kohls M Evaluate Potential Targeting Molecules. Nature Methods , 2006.

Im E-J, Lee C-H, Moon P-G, Rangaswamy GG, Lee B, Lee JM, Lee J-C, Jee J-G, Bae J-S, Kwon T-K, Kang K-W, Jeong M-S, Lee J-E, Jung H-S, Ro H-J, Jun S, Kang W, Seo S-Y, Cho Y-E, Song B-J, Baek M-C (2019) Sulfisoxazole inhibits the secretion of small extracellular vesicles by targeting the endothelin receptor A. Nature Commun 10(1):1387. doi: 10.1038/s41467-019-09387-4 PMID: 30918259

Objective: To identify a direct protein target of SFX in cancer cells.

Summary: Using data from the BindingDB, SEA identified four proteins as target candidates, which are: endothelin receptor type A, kynurenine 3-monooxygenase, carbonic anhydrase 13, and angiotensin II type 1a receptor.

Usage: Western blot (1:1000).

Related Products: Angiotensin II receptor (AT-1R) Rabbit Polyclonal, affinity-purified (Cat. #AB-N27AP)

Gulino R, Vicario N, Giunta MAS, Spoto G, Calabrese G, Vecchio M, Gulisano M, Leanza G, Parenti R (2019) Neuromuscular plasticity in a mouse neurotoxic model of spinal motoneuronal loss. Int J Mol Sci 20(6):1500. doi: 10.3390/ijms20061500

Objective: To use a neurotoxic model of spinal motoneuron depletion, induced by injection of CTB-SAP, to investigate the possible occurrence of compensatory changes in both the muscle and spinal cord.

Summary: Plastic changes in surviving motoneurons produce a functional restoration probably similar to the compensatory changes occurring in disease. These changes could be driven by glutamatergic signaling; astrocytes contacting surviving motoneurons may support this process.

Usage: Mice received 2 injections of CTB-SAP (3 mcg CTB-Sap in 2 mcL PBS) into the medial and lateral left gastrocnemius muscle.

Related Products: CTB-SAP (Cat. #IT-14)

Mouawad J, Azar JA, Bassil M (2019) Waterpipe smoke effect on oxidative stress levels in buccal cells using MAWI i-SWAB tubes. Biomark J 5(1):1-6. doi: 10.36648/2472-1646.5.1.57 PMID: 909090

Objective: To evaluate the effect of water pipe smoke on oxidative stress levels in buccal cells, using MAWI i-SWAB tubes for the detection of different protein oxidation biomarkers (S-glutathionylation, S-nitrosylation and tyrosine nitration). Furthermore, these tubes were used for the revelation of iNOS and NOX2.

Summary: Results confirmed the findings about the ability of these tubes to detect S-nitrosylation, nitrotyrosine, as well as S-glutathionylation. Results showed also the presence of immature NOX2 in human buccal cells equally.

Usage: Western blot

Related Products: NO-L-Cysteine Mouse Monoclonal, Conjugated (Cat. #AB-T125)

Eidson LN, Murphy AZ (2019) Inflammatory mediators of opioid tolerance: Implications for dependency and addiction. Peptides 115:51-58. doi: 10.1016/j.peptides.2019.01.003

Objective: To determine what mediates opioid tolerance and alterations in glutamate homeostasis.

Summary: Site-specific lesions of PAG MOR-containing neurons using Dermorphin-SAP significantly reduce the antinociceptive effects of systemic morphine suggesting that PAG MOR is critical for morphine action.

Usage: Rats were injected with 3 pmol of Dermorphin-SAP (Cat. #IT-12) into the PAG. Blank-SAP (Cat. #IT-21) was used as a control.

See: Loyd DR et al. Sex differences in micro-opioid receptor expression in the rat midbrain periaqueductal gray are essential for eliciting sex differences in morphine analgesia. J Neurosci 28:14007-14017, 2008.

Related Products: Dermorphin-SAP / MOR-SAP (Cat. #IT-12), Blank-SAP (Cat. #IT-21)

Ji J, Tao P, He L (2019) Kangxianling decoction prevents renal fibrosis in rats with 5/6 nephrectomy and inhibits Ang II-induced ECM production in glomerular mesangial cells. J Pharmacol Sci 139(4):367-372. doi: 10.1016/j.jphs.2019.03.003 PMID: 30929858

Objective: To investigate effects of Kangxianling decoction on renal fibrosis in a rat model of 5/6 Nephrectomy (5/6 N) in vivo and in angiotensin II (Ang II)-treated rat glomerular mesangial cells (HBZY-1) in vitro.

Summary: Kangxianling decoction could reduce renal fibrosis, accompanied by inhibiting the production of ECM proteins and JNK, along with downregulation of TGF-β, Ang II.

Usage: Western blot analysis (1:100).

Related Products: Angiotensin II receptor (AT-1R) Rabbit Polyclonal, affinity-purified (Cat. #AB-N27AP)