Hamster IL-1b (Interleukin-1b) ELISA Kit
- SKU:
- HMFI0011
- Product Type:
- ELISA Kit
- Size:
- 96 Assays
- Uniprot:
- Q9WVQ9
- Sensitivity:
- 9.375pg/ml
- Range:
- 15.625-1000pg/ml
- ELISA Type:
- Sandwich
- Synonyms:
- IL-1b ELISA Kit, Interleukin-1b ELISA Kit
- Reactivity:
- Hamster
Description
Hamster IL-10 ELISA
IL-1 beta is a cytokine that is mainly produced by macrophages and epithelial cells. IL-1 beta has been shown to play a role in the pathogenesis of IL-1 mediated diseases such as ankylosing spondylitis, osteoarthritis and gouty arthritis. IL-1 beta also plays a role in IL-1 mediated inflammatory diseases such as systemic IL-1 induced erythematosus (SIILE) and IL-1 induced psoriasis. The Assay Genie Hamster IL-1 beta ELISA allows researchers to accurately measure IL-1 beta in hamster serum, blood, plasma, cell culture supernatant and tissue samples.
Key Features
| Save Time | Pre-coated 96 well plate | |
| Quick Start | Kit includes all necessary reagents | |
| Publication Ready | Reproducible and reliable results |
Overview
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Product Name: |
Hamster IL-1 beta ELISA |
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Product Code: |
HMFI0011 |
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Size: |
96 Assays |
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Target: |
Hamster IL-1 beta |
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Alias: |
IL-1b, IL-1 beta |
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Reactivity: |
Hamster |
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Detection Method: |
Sandwich ELISA |
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Sensitivity: |
<9.375pg/ml |
|
Range: |
15.625-1000pg/ml |
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Storage: |
4°C for 6 months |
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Note: |
For Research Use Only |
Additional Information
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Recovery |
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Linearity: |
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Intra-Assay: |
CV <8% |
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Inter-Assay: |
CV <10% |
Protocol
*Note: Protocols are specific to each batch/lot. For the correct instructions please follow the protocol included in your kit.
| 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. |
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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. |
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5. |
Seal the plate with a cover and incubate at 37 °C for 90 min. |
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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. |
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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. |
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9. |
Remove the cover, and wash plate 3 times with Wash buffer. Let wash buffer rest in wells for 1 min between each wash. |
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10. |
Add 0.1 ml of SABC working solution into each well, cover the plate and incubate at 37°C for 30 min. |
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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. |
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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. |
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13. |
Add 50 µl of Stop solution into each well and mix thoroughly. The color changes into yellow immediately. |
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14. |
Read the O.D. absorbance at 450 nm in a microplate reader immediately after adding the stop solution. |
Sample Preparation
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. |
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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. |
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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. |
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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. |
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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 |
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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. |
Hamster IL-1b Background
IL-1b Function
IL-1B, or Interleukin-1 beta, is a vital cytokine involved in immune system functions. It activates immune cells, promotes their proliferation and differentiation, and mediates inflammation. Released by immune cells in response to infections or tissue damage, IL-1B binds to its receptor, triggering signaling pathways that activate immune cells and initiate the production of other inflammatory molecules. It also influences the fever response, stimulates acute-phase protein production, and contributes to tissue repair and remodeling.
IL-1B and Proinflammatory Effects
IL-1B is known for its prominent proinflammatory effects within the immune system. As a potent cytokine, IL-1B plays a central role in initiating and amplifying the inflammatory response. It promotes the activation of immune cells, such as macrophages and neutrophils, leading to the release of additional inflammatory mediators, including cytokines, chemokines, and prostaglandins. IL-1B also enhances the expression of adhesion molecules on endothelial cells, facilitating the migration of immune cells to the site of inflammation. Furthermore, IL-1B contributes to the development of fever, a systemic inflammatory response, by acting on the hypothalamus to increase body temperature. Overall, IL-1B's proinflammatory effects are critical for orchestrating immune responses and combating infections or tissue damage.
IL-1b Signaling Pathway
The IL-1B signaling pathway is a complex cascade of events triggered upon the binding of IL-1B to its receptor, IL-1R. Upon activation, IL-1B initiates the recruitment of an adaptor protein called MyD88, which then leads to the activation of several downstream signaling molecules. This activation ultimately results in the activation of transcription factors, such as NF-κB and AP-1, which regulate the expression of various genes involved in inflammation and immune responses. Additionally, IL-1B signaling can activate inflammasomes, multi-protein complexes that play a role in the processing and release of IL-1B itself, further amplifying the inflammatory response.
Hamster IL-1b FAQs
What is the Hamster IL-1b ELISA Kit?
The Hamster IL-1b ELISA Kit is a specialized assay kit designed to detect and quantify IL-1b levels in biological samples.
What are the advantages of using the Hamster IL-1b ELISA Kit?
The Hamster IL-1b ELISA Kit offers several advantages for researchers. It provides accurate and quantitative measurements of IL-1b levels in hamster samples, enabling precise analysis of immune responses. The kit is user-friendly, offering a straightforward procedure and providing reliable results within a short time frame. With its specificity for hamster IL-1b, researchers can confidently study IL-1b-related processes in hamsters using this kit.
Where can I find more information about the Hamster IL-1b ELISA Kit?
For more detailed information about the Hamster IL-1b, including technical specifications, performance characteristics, and ordering details, please refer to the product brochure or contact our customer support team. We are here to assist you with any inquiries you may have.
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