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Product Info Summary
| SKU: | EK0987 |
|---|---|
| Size: | 96 wells/kit, with removable strips. |
| Reactive Species: | Mouse |
| Application: | ELISA |
| Sample Types: | cell culture supernatants, cell lysates, serum and plasma (heparin, EDTA). |
Product info
Product Name
Mouse Complement C5a ELISA Kit PicoKine®
SKU/Catalog Number
EK0987
Size
96 wells/kit, with removable strips.
*Question: How many samples can I assay/run in this kit?
Description
Mouse Complement C5a ELISA Kit PicoKine® (96 Tests). Quantitate Mouse C5 in cell culture supernatants, cell lysates, serum and plasma (heparin, EDTA). Sensitivity: 10pg/ml. The brand Picokine indicates this is a premium quality ELISA kit. Each Picokine kit delivers precise quantification, high sensitivity, and excellent reproducibility. Only our most reliable and effective kits qualify as Picokine, guaranteeing top-tier results for your assays.
Storage & Handling
Store at 4°C for 6 months, at -20°C for 12 months. Avoid multiple freeze-thaw cycles (Ships with gel ice, can store for up to 3 days in room temperature. Freeze upon receiving.)
Cite This Product
Mouse Complement C5a ELISA Kit PicoKine® (Boster Biological Technology, Pleasanton CA, USA, Catalog # EK0987)
Clonality of Antibodies
See Datasheet for details
Standard Protein
Expression system for standard: E.coli; Immunogen sequence: N679-R755
Sensitivity
<10 pg/ml
Assay Range
15.6 pg/ml - 1,000 pg/ml
Standard Dilution Instructions
See datasheet of EK0987 for more details
Cross-reactivity
There is no detectable cross-reactivity with other relevant proteins.
Reactive Species
EK0987 is reactive to C5 in Mouse samples
Validated Sample Types
cell culture supernatants, cell lysates, serum and plasma (heparin, EDTA).
Application Guarantee
EK0987 is guaranteed for ELISA in Mouse by Boster Guarantee
See how Boster Bio validate our ELISA kits: ELISA Validation Information
Background of C5
C5a is a protein fragment released from complement component C5. This gene is mapped to 9q33.2. The protein encoded by this gene is the fifth component of complement, which plays an important role in inflammatory and cell killing processes. This protein is comprised of alpha and beta polypeptide chains that are linked by a disulfide bridge. An activation peptide, C5a, which is an anaphylatoxin that possesses potent spasmogenic and chemotactic activity, is derived from the alpha polypeptide via cleavage with a convertase. The C5b macromolecular cleavage product can form a complex with the C6 complement component, and this complex is the basis for formation of the membrane attack complex, which includes additional complement components. Mutations in this gene cause complement component 5 deficiency, a disease where patients show a propensity for severe recurrent infections. Defects in this gene have also been linked to susceptibility to liver fibrosis and to rheumatoid arthritis.
Kit Components
| Catalog Number | Description | Quantity |
|---|---|---|
| EK0987-CAP | Anti-Mouse C5 Pre-coated 96-well strip microplate | 1 |
| EK0987-ST | Mouse C5 Standard | 2 vials, 10 ng/tube |
| EK0987-DA | Mouse C5 Biotinylated antibody (100x) | 100ul |
| AR1103 | Avidin-Biotin-Peroxidase Complex (100x) | 100ul |
| AR1106-1 | Sample Diluent | 30ml |
| AR1106-2 | Antibody Diluent | 12ml |
| AR1106-3 | Avidin-Biotin-Peroxidase Diluent | 12ml |
| AR1104 | Color Developing Reagent (TMB) | 10ml |
| AR1105 | Stop Solution | 10ml |
| AR1106-5 | Wash Buffer (25x) | 20ml |
| PLA-SEA | Adhesive plate sealers | 4 |
*The kit components are not available for individual purchase.
Materials Required But Not Included In Kit
- Microplate Reader capable of reading absorbance at 450nm.
- Incubator.
- Automated plate washer (optional).
- Pipettes and pipette tips capable of precisely dispensing 0.5 µl through 1 ml volumes of aqueous solutions.
- Multichannel pipettes are recommended for large amount of samples.
- Deionized or distilled water.
- 500ml graduated cylinders.
- Test tubes for dilution.
Data Examples, Quality Control Data & Sample Dilution
Validation Standard Curve O.D. At 450nm
| Concentration (pg/ml) | 0 | 15.6 | 31.2 | 62.5 | 125 | 250 | 500 | 1000 |
| O.D. | 0.018 | 0.103 | 0.178 | 0.244 | 0.577 | 1.045 | 1.587 | 2.057 |
Data Example Images
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Mouse Complement C5a PicoKine ELISA Kit standard curve
Recommended Sample Dilution Ratios
According to our internal validation assays using this ELISA kit, to detect C5, Dilution ratio of 1:100, concentration in serum and plasma is around 10 ng/ml..
Intra Assay Consistency & Inter Assay Consistency
We measured random samples of Mouse Complement C5a ELISA Kit PicoKine® within the same batch/lot to ensure the consistency of the kits' performances. ELISA intra assay consistency is measured using wells from the same plate/assay kit. ELISA inter assay consistency is measured using wells from different plates from the same batch production/lot.
| Intra-Assay Precision | Inter-Assay Precision | |||||
|---|---|---|---|---|---|---|
| Sample | 1 | 2 | 3 | 1 | 2 | 3 |
| n | 16 | 16 | 16 | 24 | 24 | 24 |
| Mean (pg/ml) | 38 | 229 | 499 | 35 | 224 | 532 |
| Standard deviation | 1.78 | 16.94 | 38.92 | 2.1 | 18.81 | 49.47 |
| CV (%) | 4.7% | 7.4% | 7.8% | 6% | 8.4% | 9.3% |
Reproducibility
We ensure reproducibility by testing three samples with differing concentrations of C5 in ELISA kits from four different production batches/lots.
| Lots | Lot 1 (pg/ml) | Lot 2 (pg/ml) | Lot 3 (pg/ml) | Lot 4 (pg/ml) | Mean (pg/ml) | Standard Deviation | CV (%) |
|---|---|---|---|---|---|---|---|
| Sample 1 | 38 | 40 | 36 | 37 | 37 | 1.47 | 3.9% |
| Sample 2 | 229 | 230 | 191 | 207 | 214 | 16.26 | 7.5% |
| Sample 3 | 499 | 528 | 477 | 461 | 491 | 25.14 | 5.1% |
Protein Target Info & Infographic
Gene/Protein Information For C5 (Source: Uniprot.Org, NCBI)
Gene Name
C5
Full Name
Complement C5
Weight
188878 MW
Alternative Names
Complement C5;Hemolytic complement;Complement C5 beta chain;Complement C5 alpha chain;C5a anaphylatoxin;Complement C5 alpha' chain;C5;Hc; C5 C5Da, C5b, CPAMD4, ECLZB, C5 complement C5 complement C5|C3 and PZP-like alpha-2-macroglobulin domain-containing protein 4|C5a anaphylatoxin|anaphylatoxin C5a analog|complement component 5|prepro-C5
*if product is indicated to react with multiple species, protein info is based on the gene entry specified above in "species".For more info on C5, check out the C5 Infographic
We have 30,000+ of these available, one for each gene! check them out.
In this infographic you will see the following information for C5: database IDs, super-family, protein function, synonyms, molecular weight, chromosomal locations, tissues of expression, subcellular locations, post translational modifications, and related diseases, research areas & pathways. If you want to see more information included, or would like to contribute to it and be acknowledged, please contact us [email protected].
Specific Publications For Mouse Complement C5a ELISA Kit PicoKine® (EK0987)
Hello CJ!
EK0987 has been cited in 2 publications:
*The publications in this section are manually curated by our staff scientists. They may differ from Bioz's machine gathered results. Both are accurate. If you find a publication citing this product but is missing from this list, please let us know we will issue you a thank-you coupon.
Leukotriene B4-Mediated Neutrophil Recruitment Causes Pulmonary Capillaritis during Lethal Fungal Sepsis
Species: Mouse
The involvement of C5a in the progression of experimental arthritis with Porphyromonas gingivalis infection in SKG mice
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Customer Reviews
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Customer Q&As
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11 Customer Q&As for Mouse Complement C5a ELISA Kit PicoKine®
Question
Q: can I use citrate plasma as samples in Mouse C5 Picokine® ELISA Kit (Catalog # EK0987)?
Verified Customer
Verified customer
Asked: 2020-10-19
Answer
A: Chelating agents such as EDTA, Heparin and Citrate can bind metal ions from the functional domain of C5 causing disruption of its protein structure. C5 may be denatured as a result and may compromise the assay's measurements. The chilating sites could also be too close to the epitopes a must for detection and block the antigen antibody reaction. We have tested the C5 ELISA, treating samples with a number of anticoagulants and decided that heparin or EDTA can be used for treatment of blood/plasma samples. Do not use other anticoagulents when collecting samples.
Boster Scientific Support
Answered: 2020-10-19
Question
Q: What is the optimal O.D. value for C5 ELISA kit? I performed your C5 ELISA on serum samples. For my positive control, I received an O.D. value of 0.826, while my negative control received a value of 0.136. I obtained both of these controls from the ELISA kit, where your kit's typical data shows O.D. values much higher than my positive control and your background is lower. My samples O.D. values are around 0.225 and the highest is only 0.357. is it safe to say these samples contain C5 even though the O.D. values are not very high?
B. Roberts
Verified customer
Asked: 2020-05-06
Answer
A: The absolute O.D. values may change according to incubation time. The more you incubate the higher the O.D. values are going to be. a point of focus should be is whether your sample O.D. values are statistically significantly higher than your blank values. in the above example, you could extend your development time in the substrate incubation step to obtain higher O.D. values, as long as your negative controls' O.D. values are not increasing faster in relation to your positive controls. normally, a sample with O.D. value 2 standard deviations higher than your negative controls can be considered positive. We calculate the sensitivity of this ELISA kit by changeing cutoff O.D. value, calculated as the average of 20 negative controls plus 2 standard deviations of the 20 negative controls, into a concentration. in other words, when we claim this C5 ELISA kit to have sensitivity of 10pg/ml, that means the minimum amount of C5 that can be declared/interpreted as positive by the above standard is 10pg/ml.
Boster Scientific Support
Answered: 2020-05-06
Question
Q: if the enzyme conjugated C5 antibodies are mixed with the substrate, will that convert the substrate into the enzymatic reaction product? Or the enzyme function is only activated when the antibody is attached with the C5 antigen?
Verified Customer
Verified customer
Asked: 2020-04-18
Answer
A: The enzyme is always active. Avoid contaminating the substrate with enzyme prior to the incubation otherwise it compromises the assay with false positive signal.
Boster Scientific Support
Answered: 2020-04-18
Question
Does the antibody in this kit react with C5?
Verified Customer
Verified customer
Asked: 2020-01-08
Answer
C5 precursor is first processed by the removal of four basic residues, forming two chains (alpha and beta), linked by a disulfide bond. C5 convertase activates C5 by cleaving the alpha chain, releasing C5a anaphylatoxin. C5a is derived from proteolytic degradation of complement C5. Our C5a ELISA kit measures the natural and recombinant C5a (C5a) by itself or in the context of C5.
Boster Scientific Support
Answered: 2020-01-08
Question
What is the suggested dilution ratio for EK1400 and EK0987 in plasma samples collected from heparin tubes?
Verified Customer
Verified customer
Asked: 2019-12-09
Answer
Our suggested dilution for mouse serum when using EK1400 and EK0987 is 1:100 and the concentration range for normal serum is 6-20ng/mL.
Boster Scientific Support
Answered: 2019-12-09
Question
Q: Can C5 ELISA Kits be used with tissue homogenates (or other non-validated sample types)?
H. Green
Verified customer
Asked: 2019-10-28
Answer
A: Unfortunately, Boster Bio has not routinely validated tissue homogenates as a sample type for ELISA kits. This does not mean that ELISA kits are not suitable for other sample types than we have tested: it means further investigation is required. One will need to perform a spike and recovery study to determine if an unvalidated sample type will work with a particular kit. To perform a spike and recovery experiment, one should divide a sample into two aliquots. In one of the aliquots, the user should spike in a known amount of the kit standard. a dilution series is performed comparing the spiked versus the unspiked sample. Generally, samples with expected recovery and linearity between 80-120% are considered acceptable. This method can be used to validate any sample type that has not been assessd by Boster Bio. for a more detailed spike and recovery protocol, please contact technical support.
Note: acceptable ranges should be determined individually by each laboratory. Additionally, technical support can help determine if a buffer component is not compatible with a given ELISA kit. please view the Citations tab on the product webpage for peer-reviewed papers utilizing a wide range of sample types. We also have an innovator's reward program where if the user validates our ELISA kits in applications or samples previously not validated by Boster Bio or other users, and share such information with us by submit a review, we will reward the user's efforts with a free antibody or ELISA kit from our catalog. Biocompare.com will also give $20 Amazon giftcard as an additional reward, if the review is submitted there as well.
Boster Scientific Support
Answered: 2019-10-28
Question
Does this kit recognize uncleaved C5?
Verified Customer
Verified customer
Asked: 2019-07-25
Answer
EK0987 is able to recognize the uncleaved C5.
Boster Scientific Support
Answered: 2019-07-25
Question
Q: how do I analyze ELISA data? I have performed C5 level in serum.
Verified Customer
Verified customer
Asked: 2018-11-22
Answer
A: please read this article on ELISA data analysis. bosterbio.com/ELISA-data-analysis-instructions. we also provide a convenient online tool that you can use to analyze ELISA data. bosterbio.com/biology-research-tools/ELISA-data-analysis-online
Boster Scientific Support
Answered: 2018-11-22
Question
The product information states, “There is no detectable cross-reactivity with other relevant proteins.” Which relevant proteins were tested to have no cross-reactivity for EK0987?
Verified Customer
Verified customer
Asked: 2018-10-08
Answer
Our lab tested the following proteins and found no significant cross-reactivity or interference: <br> Recombinant mouse: Complement C3d <br> Recombinant human: Complement C3a Complement C5a
Boster Scientific Support
Answered: 2018-10-08
Question
Q: Are Boster Bio recombinant proteins and antibodies sterile?
Verified Customer
Verified customer
Asked: 2018-03-25
Answer
A: although the vials are bottled using aseptic techniques, heat-treated vials, and sterile stock solutions, they are not considered or guaranteed to be sterile. If sterile material is required for an experiment, the material can be filtered through a 0.2 micron filter designed for use with biological fluids.
Boster Scientific Support
Answered: 2018-03-25
Question
Q: how much samples can be assayed in a Picokine® ELISA Kit?
Verified Customer
Verified customer
Asked: 2014-11-22
Answer
A: The Picokine® ELISA Kits will generally run a 7-point standard curve, non-specific binding wells, and 39 samples in duplicate. this may might differ by kit so please refer to each datasheet for details.
Boster Scientific Support
Answered: 2014-11-22
