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Lps from salmonella typhosa

Manufactured by Merck Group
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LPS from Salmonella typhosa is a laboratory product available from Merck Group. It is a lipopolysaccharide extracted from the cell wall of Salmonella typhosa bacteria. The core function of this product is to serve as a reagent for research and scientific investigations.

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Lab products found in correlation

Platinum pcr supermix, by Thermo Fisher Scientific (1 mentions) Superscript 2 reverse transcriptase, by Thermo Fisher Scientific (1 mentions) Trizol, by Thermo Fisher Scientific (1 mentions) Dimethyl sulfoxide (dmso), by Merck Group (1 mentions) Anti mouse igg hrp conjugate, by Promega (1 mentions) Ab61392, by Abcam (1 mentions) Ab33915, by Abcam (1 mentions) Antibodies against nrf2, by Cell Signaling Technology (1 mentions) Sc 32233, by Santa Cruz Biotechnology (1 mentions) Gapdh, by Santa Cruz Biotechnology (1 mentions)

5 protocols using lps from salmonella typhosa

1

Quantifying B Cell Precursors

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Varying numbers of splenocytes from 4-month-old NZM or FVB mice were cultured in the presence or absence of 15 µg/ml LPS (from Salmonella typhosa; Sigma) for 7 days. Supernatants were assayed for the presence of antibodies against Hb and dsDNA (used as a positive control) by ELISA. Optical densities greater than the average of un-stimulated controls +3 SDs (for respective cell inputs) were considered indicative of the presence of antibody. Data were analyzed by Poisson analysis. Briefly, for each cell input, percentages of wells negative for the measured reactivity over the total number of wells were calculated; cell inputs resulting in a value of 37.5% were adjudged the index of the respective B cell precursor frequency.
Jain S., Bose A., Bastia B., Sharma H., Sachdeva R., Jain A.K, & Pal R. (2017). Oxidized Hemoglobin Is Antigenic and Immunogenic in Lupus. Frontiers in Immunology, 8, 732.
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2

Salmonella typhosa LPS and DMSO Assay

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LPS from Salmonella typhosa and dimethyl sulfoxide (DMSO) were purchased from Sigma-Aldrich (St. Louis, MO). SAHA was purchased from Biomol International (Plymouth Meeting, PA). Trizol, SuperScript™ II Reverse Transcriptase, and Platinum PCR SuperMix were purchased from Life Technologies (Grand Island, NY).
Zhao Y., Zhou P., Liu B., Bambakidis T., Mazitschek R., Alam H.B, & Li Y. (2014). Protective Effect of SAHA against LPS-induced Liver Damage in Rodents. The Journal of surgical research, 194(2), 544-550.
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3

Complement Pathway Modulation by RaCI2

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The effect of RaCI2 on all three pathways was tested using a Wieslab complement system screen (Euro Diagnostica, Sweden) following manufacturer’s instructions. Measurements were subtracted with buffer only and normalised for serum only samples (100% activity). Other classical and alternative pathway assays were carried out as described by A. Roos et al. 42 (link), with the following modifications. 2 μg/ml IgM (Bio-Rad) and 1.8 mg/ml LPS from Salmonella typhosa (Sigma-Aldrich) were used to coat ELISA plates. Human serum was diluted 1/100 (classical pathway) or 1/10 (alternative pathway). MAC deposition was detected with primary antibodies against C5b-9 (aE11, Abcam), and secondary anti-mouse IgG HRP conjugate (Promega). HRP activity was detected with 2,2'-Azino-bis(3-ethylbenzothiazoline-6-sulfonic acid) liquid substrate (Sigma-Aldrich) and the reaction was stopped by adding an equal volume of 1% SDS. The absorption was measured at 405nm. Measurements were subtracted with buffer only samples and normalized for serum only samples (100% activity). The competition assays were performed using size-exclusion purified inhibited C5 complexes (see purification of C5 complexes above) added to the reaction at the concentrations specified.
Jore M.M., Johnson S., Sheppard D., Barber N.M., Li Y.I., Nunn M.A., Elmlund H, & Lea S.M. (2016). Structural basis for therapeutic inhibition of complement C5. Nature structural & molecular biology, 23(5), 378-386.
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4

Quantifying Complement Activation

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Plates were coated overnight with 40 μg/ml lipopolysaccharide (LPS) from Salmonella typhosa (Sigma) at 4 °C and then blocked with 1% BSA. Plasma samples were serially diluted in GVB-MgEGTA or GVB-EDTA, added to the plate, and incubated at room temperature for 1 h. Detection of deposited C3b and plotting of the data were performed as described above.
Where indicated, CP- and AP-dependent complement activity were determined in the patient’s or a C3-depleted plasma in the absence or presence of additional normal human C3 purified as described above.
Sfyroera G., Ricklin D., Reis E.S., Chen H., Wu E.L., Kaznessis Y.N., Ekdahl K.N., Nilsson B, & Lambris J.D. (2015). Rare loss-of-function mutation in complement component C3 provides insight into molecular and pathophysiological determinants of complement activity. Journal of immunology (Baltimore, Md. : 1950), 194(7), 3305-3316.
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5

LPS-Induced Inflammatory Signaling Pathways

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LPS (from Salmonella typhosa) was purchased from Sigma (St. Louis, MO). Antibodies against TRAF6 (ab33915,1: 1000 dilution) and 3-nitrotyrosine (3-NT; ab61392,1: 300 dilution) were obtained from Abcam (Cambridge, USA). Antibodies against transforming growth factor-β (TGF-β; sc-146, 1: 300 dilution), collagen (col) IV (sc-29010,1: 300 dilution), Bax (sc-7480,1: 300 dilution), Bcl-2 (sc-7382,1: 300 dilution), IκB-α(sc-373893,1: 300 dilution), heme oxygenase-1 (HO-1; sc-136960,1: 300 dilution), GAPDH (sc-32233,1: 300 dilution) and the secondary horseradish peroxidase-conjugated antibody (7074, 1: 5000 dilution) were obtained from Santa Cruz Biotechnology. Antibodies against Nrf2 was obtained from Cell Signaling Technology (CST, CA, USA).
Chen X., Zhao Y., Xu J., Bao J., Zhao J., Chen J., Chen G, & Han J. (2020). The Nephroprotective Effect of TNF Receptor-Associated Factor 6 (TRAF6) Blockade on LPS-Induced Acute Renal Injury Through the Inhibition if Inflammation and Oxidative Stress. Medical Science Monitor : International Medical Journal of Experimental and Clinical Research, 26, e919698-1-e919698-12.
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