Human complement C5a ELISA Kit PicoKine®

C5 ELISA kit for Human

Human complement C5a ELISA Kit PicoKine™ (96 Tests). Quantitate Human C5 in cell culture supernatants, cell lysates, serum and plasma (heparin, EDTA). Sensitivity: 10pg/ml. Cited in 1 publication(s).

Product Info Summary

SKU: EK0966
Size: 96 wells/kit, with removable strips.
Reactive Species: Human
Application: ELISA
Sample Types: cell culture supernatants, cell lysates, serum and plasma (heparin, EDTA).

Product Name

Human complement C5a ELISA Kit PicoKine®

View all C5 ELISA kits

SKU/Catalog Number

EK0966

Size

96 wells/kit, with removable strips.

*Question: How many samples can I assay/run in this kit?

Description

Human complement C5a ELISA Kit PicoKine™ (96 Tests). Quantitate Human C5 in cell culture supernatants, cell lysates, serum and plasma (heparin, EDTA). Sensitivity: 10pg/ml.

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

Human complement C5a ELISA Kit PicoKine® (Boster Biological Technology, Pleasanton CA, USA, Catalog # EK0966)

Clonality of Antibodies

See Datasheet for details

Immunogen

Expression system for standard: E.coli; Immunogen sequence: T678-R751

Sensitivity

<10 pg/ml

Assay Range

31.2 pg/ml - 2,000 pg/ml

Standard Dilution Instructions

serial dilution instructions image

Add 100ul of sample diluent in well #2-#8. Add 200ul standard stock solution to well #1, and serial dilute for well #2-#7 to make a standard curve row. Leave well #8 as blank See datasheet of EK0966 for more details

Cross-reactivity

There is no detectable cross-reactivity with other relevant proteins.

Reactive Species

EK0966 is reactive to C5 in Human samples

Validated Sample Types

cell culture supernatants, cell lysates, serum and plasma (heparin, EDTA).

Application Guarantee

EK0966 is guaranteed for ELISA in Human 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
EK0966-CAP Anti-Human C5 Pre-coated 96-well strip microplate 1
EK0966-ST Human C5 Standard 2 vials, 10 ng/tube
EK0966-DA Human 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.

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Validation Standard Curve O.D. At 450nm

Concentration (pg/ml)031.262.512525050010002000
O.D.0.0740.1870.2680.4770.6900.9401.3921.930

Data Example Images

Recommended Sample Dilution Ratios

According to our internal validation assays using this ELISA kit, to detect C5, Dilution ratio of 1:200, concentration in serum and plasma is around 80 ng/ml..

Intra Assay Consistency & Inter Assay Consistency

We measured random samples of Human 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 PrecisionInter-Assay Precision
Sample123123
n161616242424
Mean (pg/ml)892851037842851015
Standard deviation4.0917.167.466.8221.0969.02
CV (%)4.6%6%6.4%6.1%7.4%6.8%

Reproducibility

We ensure reproducibility by testing three samples with differing concentrations of C5 in ELISA kits from four different production batches/lots.

LotsLot 1 (pg/ml)Lot 2 (pg/ml)Lot 3 (pg/ml)Lot 4 (pg/ml)Mean (pg/ml)Standard DeviationCV (%)
Sample 189979884925.786.2%
Sample 228526125625826511.684.4%
Sample 31037990926101099040.934.1%
*number of samples for each test n=16.

Gene/Protein Information For C5 (Source: Uniprot.Org, NCBI)

Gene Name

C5

Full Name

Complement C5

Weight

188.305kDa

Alternative Names

C5; Complement Component C5/C5a 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

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].

Hello CJ!

EK0966 has been cited in 1 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.

Effect of preoperative inhaled budesonide on pulmonary injury after cardiopulmonary bypass: A randomized pilot study

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11 Customer Q&As for Human complement C5a ELISA Kit PicoKine®

Question

Q: is it okay to use citrate plasma as samples in Human C5 Picokine® ELISA Kit (Catalog # EK0966)?

Verified Customer

Verified customer

Asked: 2020-09-23

Answer

A: Chelating agents such as EDTA, Heparin and Citrate can sequester 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 required for detection and block the antigen antibody reaction. We have tested the C5 ELISA, treating samples with various 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: 2020-09-23

Question

Q: how long can samples (cell cultures, serum, and plasma) be stored and still be stable for measuring C5 using the EK0966 Human C5 Picokine® ELISA Kit?

Verified Customer

Verified customer

Asked: 2020-02-04

Answer

A: Boster Bio does not assess sample stability. Variations in sample collection, processing, and storage may change the stabilityof samples. It is recommend to assay sample on-stat collection when possible, or aliquot into single use volumes and store samples frozen. limit repetitive freeze-thaw cycles with the stored samples to prevent protein degradation.

Boster Scientific Support

Answered: 2020-02-04

Question

Q: if the enzyme conjugated C5 antibodies are mixed with the substrate, will that change the substrate into the enzymatic reaction product? Or the enzyme function is only activated when the antibody is attached with the C5 antigen?

E. Puri

Verified customer

Asked: 2019-12-31

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: 2019-12-31

Question

Q: how do I analyze ELISA data? I have performed C5 level in serum.

K. Babu

Verified customer

Asked: 2019-11-10

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: 2019-11-10

Question

A customer is asking if EK0966 wiil measure C5a or C5a-desArg, or both?

Verified Customer

Verified customer

Asked: 2019-10-25

Answer

EK0966 measures C5a only.

Boster Scientific Support

Answered: 2019-10-25

Question

Q: what is the protocol regarding preparation of cell lysates prepared for use in Picokine® ELISA kits?

Verified Customer

Verified customer

Asked: 2019-09-25

Answer

A: in those Picokine® ELISAs where cell or tissue lysate is a validated sample type, sample preparation instructions for lysate can be found in the product insert. Components in lysate and lysis buffer can effect immunoreactivity, so if lysate is not a validated sample type, care must be taken in sample preparation and validation.

Boster Scientific Support

Answered: 2019-09-25

Question

Q: Are Boster Bio recombinant proteins and antibodies sterile?

Verified Customer

Verified customer

Asked: 2019-07-06

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: 2019-07-06

Question

I wanted to know if EK0966 measures C5a or C5a-desArg, or both?

Verified Customer

Verified customer

Asked: 2018-02-28

Answer

EK0966 only measures C5a.

Boster Scientific Support

Answered: 2018-02-28

Question

Q: is there any online tool I can use to streamline the data analysis for my ELISA results?

J. Miller

Verified customer

Asked: 2016-04-15

Answer

A: We have a web based ELISA curve fitting (4pl) and data analysis tool. Please give it a try: bosterbio.com/biology-research-tools/ELISA-data-analysis-online. You can also consult our article on ELISA data analysis: bosterbio.com/ELISA-data-analysis-instructions

Boster Scientific Support

Answered: 2016-04-15

Question

Q: can you suggest the dilution ratio of serum samples for detection of C5 in Human serum? I am trying to measure a a number of analytes and it requires 100ul of diluted samples for each well. We have limited sample volumes so we like to dilute as much as possible.

Verified Customer

Verified customer

Asked: 2016-03-11

Answer

A: without knowing the physiological or pathological context of your samples we cannot recommend a dilution ratio without performing a pilot test with your samples. Here is how you can perform a pilot study on your own: perform a serial dilution of your samples on the C5 ELISA kit to make sure you have a linear ascending curve followed by a plateau, which signifies the samples saturating the detection limit of the kit. Then you can pick the dilution ratios from samples in the linear part of the curve as your experimental dilution ratio.
If you are interested in using our ELISA service, you can also send us your sample and we will take care of everything for you. You can check our service details here: bosterbio.com/services/assay-services/ELISA-testing-service
Since you mentioned you have limited samples, our cost effective multiplex ELISA service would fit perfectly for your needs, where we can generate dozens of data points using as little as 25ul sample volume. Information on this service is also in the above link.

Boster Scientific Support

Answered: 2016-03-11

Question

Q: how much samples can be assayed in a Picokine® ELISA Kit?

Verified Customer

Verified customer

Asked: 2015-08-22

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: 2015-08-22

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