Human S100B / S100 beta ELISA Kit
- SKU:
- HUFI00390
- Product Type:
- ELISA Kit
- Size:
- 96 Assays
- Uniprot:
- P04271
- Sensitivity:
- 18.75pg/ml
- Range:
- 31.25-2000pg/ml
- ELISA Type:
- Sandwich
- Synonyms:
- S100B, Protein S100-B, S100, S100 beta, S100 calcium binding protein B, S100 calcium-binding protein B, S100 calcium-binding protein, beta, neural, S-100 calcium-binding protein, beta chain, 10protein S100-B, S-100 protein beta chain, S-100 protein s
- Reactivity:
- Human
- Research Area:
- Cell Biology
Description
Human S100B / S100 beta ELISA Kit
S100B (S100 Calcium Binding Protein B) encodes an EF-hand type Ca(2+)-binding protein with two calcium-binding motifs at very similar locations in the sequence, suggesting that each may bind a single calcium ion. Studies using human recombinant calreticulin suggest that it can function as both an intracellular chaperone for nascent polypeptides and as a cytosolic receptor for bacterial lipoproteins, such as displaced apolipoprotein J (apoJ) from Corynebacterium diphtheriae. Pathways related with S100B include Transport to the Golgi and subsequent modification. An important paralog of S100Bis S100A8.
Key Features
| Save Time | Pre-coated 96 well plate | |
| Quick Start | Kit includes all necessary reagents | |
| Publication Ready | Reproducible and reliable results |
Overview
|
Product Name: |
Human S100B / S100 beta ELISA Kit |
|
Product Code: |
HUFI00390 |
|
Size: |
96 Assays |
|
Alias: |
S100B, Protein S100-B, S100, S100 beta, S100 calcium binding protein B, S100 calcium-binding protein B, S100 calcium-binding protein, beta, neural, S-100 calcium-binding protein, beta chain, 10protein S100-B, S-100 protein beta chain, S-100 protein subunit beta, S100beta |
|
Detection Method: |
Sandwich ELISA, Double Antibody |
|
Application: |
This immunoassay kit allows for the in vitro quantitative determination of Human S100B concentrations in serum plasma and other biological fluids. |
|
Sensitivity: |
18.75pg/ml |
|
Range: |
31.25-2000pg/ml |
|
Storage: |
4°C for 6 months |
|
Note: |
For Research Use Only |
Additional Information
|
Recovery |
Matrices listed below were spiked with certain level of Human S100B and the recovery rates were calculated by comparing the measured value to the expected amount of Human S100B in samples.
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Linearity |
The linearity of the kit was assayed by testing samples spiked with appropriate concentration of [ANALYTE] and their serial dilutions. The results were demonstrated by the percentage of calculated concentration to the expected.
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CV(%) |
Intra-Assay: CV<8% |
Kit Components
| Component | Quantity | Storage |
|
ELISA Microplate (Dismountable) |
8x12 strips |
4°C for 6 months |
|
Lyophilized Standard |
2 |
4°C/ -20°C |
|
Sample/Standard Dlution Buffer |
20ml |
4°C |
|
Biotin-labeled Antibody (Concentrated) |
120ul |
4°C (Protection from light) |
|
Antibody Dilution Buffer |
10ml |
4°C |
|
HRP-Streptavidin Conjugate (SABC) |
120ul |
4°C (Protect from light) |
|
SABC Dilution Buffer |
10ml |
4°C |
|
TMB Substrate |
10ml |
4°C (Protection from light) |
|
Stop Solution |
10ml |
4°C |
|
Wash Buffer (25X) |
30ml |
4°C |
|
Plate Sealer |
5 |
- |
Other materials 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
Protein Information
Protocol
*Note: Protocols are specific to each batch/lot. For the exact 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.
| Step | Procedure |
|
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. |
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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. |
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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. |
Sample Type
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 Type | Protocol |
|
Serum |
If 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. |
|
Plasma |
Collect 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 Fluid |
Collect 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 supernatant |
Collect 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 lysates |
Solubilize 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 homogenates |
The 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 lysates |
Rinse 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 Milk |
Collect 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. |
S100B Background
S100B Background
S100B is a protein that plays a crucial role in various biological processes within the human body. It belongs to the S100 protein family, which encompasses a group of calcium-binding proteins found primarily in the central nervous system (CNS) and peripheral tissues. S100B is predominantly expressed by astrocytes, a type of glial cell in the CNS, and it serves as an important marker for astrocytic activity. Additionally, S100B is involved in regulating intracellular calcium levels, cell proliferation, differentiation, and cell survival. Its elevated levels in the bloodstream have been associated with several neurological conditions, including brain injuries, neurodegenerative diseases, and psychiatric disorders.
S100B as a Marker
S100B is a biomarker primarily generated by astrocytes in the central nervous system (CNS), indicating astrocytic activation. Research employing immunohistochemical techniques has revealed that S100B-positive cells are predominantly astrocytes in gray matter and oligodendrocytes in white matter. In numerous instances of brain injury, both intra- and extracellular S100B mRNA and protein levels have been utilized to gauge astrocyte activation and/or death. Increased concentrations of S100B in serum or cerebrospinal fluid (CSF) have been associated with various CNS disorders. Although S100B release may often be an effect rather than the cause of the condition, it strongly suggests that S100B has significant potential as a marker for CNS injury.
S100B Gene
S100B is a protein characterized by its homodimeric structure, with each beta monomer weighing approximately 10.5 kDa. Similar to other S100 proteins, S100B undergoes a significant conformational change upon binding calcium, allowing hydrophobic residues to become exposed and interact with other proteins, thereby facilitating its biological activity. This protein is primarily found in the cytoplasm and nucleus of astrocytes, along with other members of the S100 family. Its presence in these cells contributes to the regulation of cytoskeletal structure and cell proliferation. Although S100B is predominantly associated with astroglial and Schwann cells, it has also been identified in various other cell types such as adipocytes, chondrocytes, lymphocytes, bone marrow cells, and melanocytes. Elimination of S100B primarily occurs through renal excretion.
S100B ELISA Kit FAQs
What is the S100B ELISA Kit used for?
The S100B ELISA kit specifically targets the S100B protein, which is primarily expressed by astrocytes in the central nervous system (CNS). Elevated levels of S100B have been associated with various neurological conditions, including brain injuries, neurodegenerative diseases, and psychiatric disorders.
What are the advantages of using the S100B ELISA Kit?
The S100B ELISA Kit offers several advantages, including high sensitivity, accuracy, and reproducibility. It provides a user-friendly and reliable method to quantify S100B levels in biological specimens, allowing for precise measurements and robust data analysis.
What sample types are compatible with S100B ELISA Kit?
The S100B ELISA Kit is compatible with various sample types, including serum, plasma, cell lysates, and tissue homogenates. It provides flexibility in sample selection, allowing researchers to analyze S100B levels in different biological matrices.
What are the storage requirements with S100B ELISA Kit?
The S100B ELISA Kit components should be stored according to the instructions provided in the kit manual. Generally, it is recommended to store the kit components at the recommended temperature to ensure their stability and optimal performance.
What should I do if my assay results are not optimal?
If you encounter any issues or have suboptimal assay results, we recommend contacting our dedicated support team for assistance. They will be available to provide troubleshooting guidance, answer your questions, and ensure you achieve the best possible results with the S100B ELISA Kit.
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