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Human Calponin-2 / CNN2 ELISA Kit

SKU:
HUFI01273
Product Type:
ELISA Kit
Size:
96 Assays
Uniprot:
Q99439
Sensitivity:
0.469ng/ml
Range:
0.781-50ng/ml
ELISA Type:
Sandwich
Synonyms:
CNN2, Calponin-2, Calponin H2, smooth muscle, Neutral calponin
Reactivity:
Human
Research Area:
Cell Biology
€599
Frequently bought together:

Description

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Human Calponin-2/CNN2 ELISA Kit

The Human Calponin-2 (CNN2) ELISA Kit is specifically designed for the quantitative detection of calponin-2 levels in human serum, plasma, and cell culture supernatants. This kit offers exceptional sensitivity and specificity, allowing for accurate and reproducible results that are crucial for various research applications.Calponin-2, also known as CNN2, is a key protein involved in smooth muscle contraction and cytoskeletal organization. Its expression has been linked to conditions such as vascular diseases, smooth muscle-related disorders, and cancer progression.

Therefore, accurate measurement of calponin-2 levels is essential for studying these conditions and identifying potential therapeutic targets.Overall, the Human Calponin-2 (CNN2) ELISA Kit is a valuable tool for researchers looking to investigate the role of calponin-2 in various disease states and develop novel treatment strategies. Its high sensitivity and reliability make it a suitable choice for a wide range of research projects in the field of biology and medicine.

Product Name:Human Calponin-2 / CNN2 ELISA Kit
Product Code:HUFI01273
Size:96 Assays
Alias:CNN2, Calponin-2, Calponin H2, smooth muscle, Neutral calponin
Detection method:Sandwich ELISA, Double Antibody
Application:This immunoassay kit allows for the in vitro quantitative determination of Human CNN2 concentrations in serum plasma and other biological fluids.
Sensitivity:0.469ng/ml
Range:0.781-50ng/ml
Storage:4°C for 6 months
Note:For Research Use Only
Recovery:Matrices listed below were spiked with certain level of Human CNN2 and the recovery rates were calculated by comparing the measured value to the expected amount of Human CNN2 in samples.
MatrixRecovery range(%)Average(%)
serum(n=5)87-10498
EDTA plasma(n=5)86-9791
UFH plasma(n=5)87-10498
Linearity:The linearity of the kit was assayed by testing samples spiked with appropriate concentration of Human CNN2 and their serial dilutions. The results were demonstrated by the percentage of calculated concentration to the expected.
Sample1:21:41:8
serum(n=5)89-105%88-103%87-98%
EDTA plasma(n=5)88-101%92-99%83-100%
UFH plasma(n=5)84-99%87-99%81-94%
CV(%):Intra-Assay: CV<8%
Inter-Assay: CV<10%
ComponentQuantityStorage
ELISA Microplate (Dismountable)8×12 strips4°C for 6 months
Lyophilized Standard24°C/-20°C
Sample/Standard Dilution Buffer20ml4°C
Biotin-labeled Antibody(Concentrated)120ul4°C (Protect from light)
Antibody Dilution Buffer10ml4°C
HRP-Streptavidin Conjugate(SABC)120ul4°C (Protect from light)
SABC Dilution Buffer10ml4°C
TMB Substrate10ml4°C (Protect from light)
Stop Solution10ml4°C
Wash Buffer(25X)30ml4°C
Plate Sealer5-

Other materials and equipment required:

  • Microplate reader with 450 nm wavelength filter
  • Multichannel Pipette, Pipette, microcentrifuge tubes and disposable pipette tips
  • Incubator
  • Deionized or distilled water
  • Absorbent paper
  • Buffer resevoir
UniprotQ99439
UniProt Protein Function:calponin 2: Thin filament-associated protein that is implicated in the regulation and modulation of smooth muscle contraction. It is capable of binding to actin, calmodulin, troponin C and tropomyosin. The interaction of calponin with actin inhibits the actomyosin Mg-ATPase activity. Belongs to the calponin family. 2 isoforms of the human protein are produced by alternative splicing.
UniProt Protein Details:Protein type:Actin-binding; Motility/polarity/chemotaxis

Chromosomal Location of Human Ortholog: 19p13.3

Cellular Component: cytoskeleton; focal adhesion; intercellular junction; membrane; stress fiber

Biological Process: cytoskeleton organization and biogenesis; regulation of actin filament-based process

NCBI Summary:The protein encoded by this gene, which can bind actin, calmodulin, troponin C, and tropomyosin, may function in the structural organization of actin filaments. The encoded protein could play a role in smooth muscle contraction and cell adhesion. Several pseudogenes of this gene have been identified, and are present on chromosomes 1, 2, 3, 6, 9, 11, 13, 15, 16, 21 and 22. Alternative splicing results in multiple transcript variants encoding different isoforms. [provided by RefSeq, Jan 2015]
UniProt Code:Q99439
NCBI GenInfo Identifier:6226844
NCBI Gene ID:1265
NCBI Accession:Q99439.4
UniProt Secondary Accession:Q99439,Q92578, A5D8U8, A6NFI4, D6W5X9,
UniProt Related Accession:Q99439
Molecular Weight:29,500 Da
NCBI Full Name:Calponin-2
NCBI Synonym Full Names:calponin 2
NCBI Official Symbol:CNN2
NCBI Protein Information:calponin-2
UniProt Protein Name:Calponin-2
UniProt Synonym Protein Names:Calponin H2, smooth muscle; Neutral calponin
Protein Family:Calponin
UniProt Gene Name:CNN2
UniProt Entry Name:CNN2_HUMAN

*Note: Protocols are specific to each batch/lot. For the correct instructions please follow the protocol included in your kit.

Before adding to wells, equilibrate the SABC working solution and TMB substrate for at least 30 min at 37°C. When diluting samples and reagents, they must be mixed completely and evenly. It is recommended to plot a standard curve for each test.

StepProtocol
1.Set standard, test sample and control (zero) wells on the pre-coated plate respectively, and then, record their positions. It is recommended to measure each standard and sample in duplicate. Wash plate 2 times before adding standard, sample and control (zero) wells!
2.Aliquot 0.1ml standard solutions into the standard wells.
3.Add 0.1 ml of Sample / Standard dilution buffer into the control (zero) well.
4.Add 0.1 ml of properly diluted sample ( Human serum, plasma, tissue homogenates and other biological fluids.) into test sample wells.
5.Seal the plate with a cover and incubate at 37 °C for 90 min.
6.Remove the cover and discard the plate content, clap the plate on the absorbent filter papers or other absorbent material. Do NOT let the wells completely dry at any time. Wash plate X2.
7.Add 0.1 ml of Biotin- detection antibody working solution into the above wells (standard, test sample & zero wells). Add the solution at the bottom of each well without touching the side wall.
8.Seal the plate with a cover and incubate at 37°C for 60 min.
9.Remove the cover, and wash plate 3 times with Wash buffer. Let wash buffer rest in wells for 1 min between each wash.
10.Add 0.1 ml of SABC working solution into each well, cover the plate and incubate at 37°C for 30 min.
11.Remove the cover and wash plate 5 times with Wash buffer, and each time let the wash buffer stay in the wells for 1-2 min.
12.Add 90 µl of TMB substrate into each well, cover the plate and incubate at 37°C in dark within 10-20 min. (Note: This incubation time is for reference use only, the optimal time should be determined by end user.) And the shades of blue can be seen in the first 3-4 wells (with most concentrated standard solutions), the other wells show no obvious color.
13.Add 50 µl of Stop solution into each well and mix thoroughly. The color changes into yellow immediately.
14. Read the O.D. absorbance at 450 nm in a microplate reader immediately after adding the stop solution.

When carrying out an ELISA assay it is important to prepare your samples in order to achieve the best possible results. Below we have a list of procedures for the preparation of samples for different sample types.

Sample TypeProtocol
SerumIf using serum separator tubes, allow samples to clot for 30 minutes at room temperature. Centrifuge for 10 minutes at 1,000x g. Collect the serum fraction and assay promptly or aliquot and store the samples at -80°C. Avoid multiple freeze-thaw cycles.

If serum separator tubes are not being used, allow samples to clot overnight at 2-8°C. Centrifuge for 10 minutes at 1,000x g. Remove serum and assay promptly or aliquot and store the samples at -80°C. Avoid multiple freeze-thaw cycles.

PlasmaCollect plasma using EDTA or heparin as an anticoagulant. Centrifuge samples at 4°C for 15 mins at 1000 × g within 30 mins of collection. Collect the plasma fraction and assay promptly or aliquot and store the samples at -80°C. Avoid multiple freeze-thaw cycles. Note: Over haemolysed samples are not suitable for use with this kit.
Urine & Cerebrospinal FluidCollect the urine (mid-stream) in a sterile container, centrifuge for 20 mins at 2000-3000 rpm. Remove supernatant and assay immediately. If any precipitation is detected, repeat the centrifugation step. A similar protocol can be used for cerebrospinal fluid.
Cell culture supernatantCollect the cell culture media by pipette, followed by centrifugation at 4°C for 20 mins at 1500 rpm. Collect the clear supernatant and assay immediately.
Cell lysatesSolubilize cells in lysis buffer and allow to sit on ice for 30 minutes. Centrifuge tubes at 14,000 x g for 5 minutes to remove insoluble material. Aliquot the supernatant into a new tube and discard the remaining whole cell extract. Quantify total protein concentration using a total protein assay. Assay immediately or aliquot and store at ≤ -20 °C.
Tissue homogenatesThe preparation of tissue homogenates will vary depending upon tissue type. Rinse tissue with 1X PBS to remove excess blood & homogenize in 20ml of 1X PBS (including protease inhibitors) and store overnight at ≤ -20°C. Two freeze-thaw cycles are required to break the cell membranes. To further disrupt the cell membranes you can sonicate the samples. Centrifuge homogenates for 5 mins at 5000xg. Remove the supernatant and assay immediately or aliquot and store at -20°C or -80°C.
Tissue lysatesRinse tissue with PBS, cut into 1-2 mm pieces, and homogenize with a tissue homogenizer in PBS. Add an equal volume of RIPA buffer containing protease inhibitors and lyse tissues at room temperature for 30 minutes with gentle agitation. Centrifuge to remove debris. Quantify total protein concentration using a total protein assay. Assay immediately or aliquot and store at ≤ -20 °C.
Breast MilkCollect milk samples and centrifuge at 10,000 x g for 60 min at 4°C. Aliquot the supernatant and assay. For long term use, store samples at -80°C. Minimize freeze/thaw cycles.

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