Active Complement Component 5a (C5a) Homo sapiens (Human) Active protein

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Overview
Properties
  • Buffer FormulationPBS, pH7.4, containing 5% Trehalose.
  • Traits Freeze-dried powder, Purity > 95%
  • Isoelectric Point8.9
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  • Active Complement Component 5a (C5a) Packages (Simulation)
  • Active Complement Component 5a (C5a) Packages (Simulation)
  • APA388Hu61.png Figure. SDS-PAGE
  • Active Complement Component 5a (C5a) WB Image
  • Certificate ISO9001: 2008, ISO13485: 2003 Registered

Activity test

Complement Component 5a (C5a) is a component of the complement system which plays a key role in promoting migration and adherence of neutrophils and monocytes to vessel walls. C5a has been proven to be able to induce chemotactic migration of THP-1 cells. Therefore, chemotaxis assay used 24-well microchemotaxis system was undertaken to detect the chemotactic effect of C5a on the human monocytic cell line THP-1. Briefly, THP-1 cells were seeded into the upper chambers (100µL cell suspension, 106 cells/mL in RPMI 1640 with 0.5% FBS) and C5a (50ng/mL and 100ng/mL diluted separately in serum free RPMI 1640) was added in lower chamber with a polycarbonate filter (8µm pore size) used to separate the two compartments. After incubation at 37oC with 5% CO2 for 2h, the filter was removed, then cells in low chamber were observed by inverted microscope at low magnification (×100) and the number of migrated cells were counted at high magnification (×400) randomly (five fields for each filter). Result: C5a is able to induce migration of THP-1 cells. The migrated THP-1 cells in low chamber at low magnification (×100) were shown in Figure 1. Five fields of each chamber were randomly chosen to count the migrated cells at high magnification (×400) and the statistical data was shown in Figure 2.
(A) THP-1 cells were seeded into the upper chambers and 50ng/mL C5a was added in lower chamber, then cells in lower chamber were observed at low magnification (×100) after incubation for 3h;
(B) THP-1 cells were seeded into the upper chambers and 100ng/mL C5a was added in lower chamber, then cells in lower chamber were observed at low magnification (×100) after incubation for 3h;
(C) THP-1 cells were seeded into the upper chambers and serum free RPMI 1640 without C5a was added in lower chamber, then cells in lower chamber were observed at low magnification (×100) after incubation for 3h.
Figure. The chemotactic effect of C5a on THP-1 cells.

Figure. The chemotactic effect of C5a on THP-1 cells

The activity of recombinant human C5a was also measured by its ability to induce N-acetyl-β-D-glucosaminidase release from differentiated U937 human histiocytic lymphoma cells. 3.2*106 differentiated U937 cells were added to the 24-well plate and different concentrations of rhC5a was added to the 24-well plate and incubated at 37 ℃ for 3min. The cells were centrifuged at 400 g for 3min, and the supernatant contained N-acetyl-β-D-glucosaminidase. The enzyme activity of N-acetyl-β-D-glucosaminidase was measured by the substrate of 4-Nitrophenyl 2-acetamido-2-deoxy-β-D-glucopyranoside. The result was shown in figure 1, It was obvious that rhC5a can significantly induce N-acetyl-β-D-glucosaminidase release.

Usage

Reconstitute in 10mM PBS (pH7.4) to a concentration of 0.1-1.0 mg/mL. Do not vortex.

Storage

Avoid repeated freeze/thaw cycles. Store at 2-8°C for one month. Aliquot and store at -80°C for 12 months.

Stability

The thermal stability is described by the loss rate. The loss rate was determined by accelerated thermal degradation test, that is, incubate the protein at 37°C for 48h, and no obvious degradation and precipitation were observed. The loss rate is less than 5% within the expiration date under appropriate storage condition.

Citations

  • Cathepsin D is released after severe tissue trauma in vivo and is capable of generating C5a in vitroScienceDirect: S0161589011008297
  • Elevated complement factor C5a in maternal and umbilical cord plasma in preeclampsiaPubMed: 23415845
  • Complement component 5 contributes to poor disease outcome in humans and mice with pneumococcal meningitisPubMed: PMC3195471
  • Efficacy of a synthetic antimicrobial peptidomimetic versus vancomycin in a Staphylococcus epidermidis device-related murine peritonitis modelPubmed: 23645587
  • Conversion from external fixator to intramedullary nail causes a second hit and impairs fracture healing in a severe trauma modelPubmed: 23070742
  • Therapeutic effects of sesame oil on monosodium urate crystal-induced acute inflammatory response in ratsPubmed: 24353977
  • Evaluation of the Markers of Inflammation in the Umbilical Cord Blood of Newborns of Mothers with ThrombophiliaPubmed:25209155
  • Complement Split Products C3a/C5a and Receptors: Are They Regulated by Circulating Angiotensin II Type 1 Receptor Autoantibody in Severe Preeclampsia?PubMed: 26485247
  • Acute and prolonged complement activation in the central nervous system during herpes simplex encephalitisPubmed:27235358
  • Consistency and pathophysiological characterization of a rat polymicrobial sepsis model via the improved cecal ligation and puncture surgeryPubmed:26802602
  • Dosagem de frações ativadas do sistema complemento em empiema induzido em ratos10183
  • Cadmium-induced immune abnormality is a key pathogenic event in human and rat models of preeclampsiapubmed:27511439
  • Complement C5a/C5aR pathway potentiates the pathogenesis Q5 of gastric cancer by down-regulating p21 expressionpubmed:29031586
  • Serum complement factor 5a levels are associated with nonalcoholic fatty liver disease in obese childrenDOI: 10.1111/apa.14106
  • Splenectomy modulates early immuno-inflammatory responses to trauma-hemorrhage and protects mice against secondary sepsisPubmed: 30291296
  • The Beneficial Effects of Lactobacillus plantarum PS128 on High-Intensity, Exercise-Induced Oxidative Stress, Inflammation, and Performance in TriathletesPubmed: 30736479
  • Intrathecal complement activation by the classical pathway in tick-borne encephalitisPubmed: 30850976
  • Sepsis causes right ventricular myocardial inflammation independent of pulmonary hypertension in a porcine sepsis modelPubmed: 31247004
  • C5a/C5aR Pathway is Critical for the Pathogenesis of PsoriasisPubmed: 31447855
  • Novel evidence that an alternative complement cascade pathway is involved in optimal mobilization of hematopoietic stem/progenitor cells in Nlrp3 …
  • C5aR deficiency attenuates the breast cancer development via the p38/p21 axisPubmed: 32669478
  • A Novel Evidence That Mannan Binding Lectin (MBL) Pathway of Complement Cascade Activation is Involved in Homing and Engraftment of Hematopoietic Stem …Pubmed: 32406006
  • Modulation of C5a-C5aR interactions against murine mammary cancer cell line
  • C‐reactive protein inhibits C3a/C3aR‐dependent podocyte autophagy in favor of diabetic kidney diseasePubmed:35503088

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