Human HBXIP / Hepatitis B virus X-interacting protein ELISA Kit
- SKU:
- HUFI01203
- Product Type:
- ELISA Kit
- Size:
- 96 Assays
- Uniprot:
- O43504
- Sensitivity:
- 0.938ng/ml
- Range:
- 1.563-100ng/ml
- ELISA Type:
- Sandwich
- Synonyms:
- HBXIP, Hepatitis B virus X-interacting protein, HBV X-interacting protein, HBX-interacting protein, MGC71071, XIP
- Reactivity:
- Human
- Research Area:
- Cell Biology
Description
Human HBXIP / Hepatitis B virus X-interacting protein ELISA Kit
HBXIP is important in viral replication, as it binds to the C-terminus of hepatitis B virus X protein (HBx). This protein's work is to suppress HBx activity and thus change the virus' replication cycle. HBXIP is associated with several diseases, including viral hepatitis B and C. RET signaling and signaling by GPCR are among HBXIP related pathways. The Assay Genie HBXIP ELISA is a highly sensitive assay for the quantitative measurement of HBXIP in serum, 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
|
Product Name: |
Human HBXIP / Hepatitis B virus X-interacting protein ELISA Kit |
|
Product Code: |
HUFI01203 |
|
Size: |
96 Assays |
|
Alias: |
HBXIP, Hepatitis B virus X-interacting protein, HBV X-interacting protein, HBX-interacting protein, MGC71071, XIP |
|
Detection Method: |
Sandwich ELISA, Double Antibody |
|
Application: |
This immunoassay kit allows for the in vitro quantitative determination of Human HBXIP concentrations in serum plasma and other biological fluids. |
|
Sensitivity: |
0.938ng/ml |
|
Range: |
1.56-100ng/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 HBXIP and the recovery rates were calculated by comparing the measured value to the expected amount of Human HBXIP in samples.
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Linearity |
The linearity of the kit was assayed by testing samples spiked with appropriate concentration of Human HBXIP 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. |
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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. |
|
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. |
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. |
Hepatitis B virus X-interacting Protein Background
Hepatitis B
Hepatitis B is a viral infection that primarily affects the liver. It is caused by the hepatitis B virus (HBV) and is a major global health concern. HBV is transmitted through contact with the blood or other body fluids of an infected person, such as through unprotected sexual intercourse, sharing needles, or from an infected mother to her newborn during childbirth. The infection can be acute, meaning it lasts for a short period, or it can become chronic, lasting for more than six months. Chronic hepatitis B can lead to serious complications, including liver cirrhosis, liver failure, and an increased risk of liver cancer. Timely diagnosis and monitoring of hepatitis B are crucial for effective management and treatment strategies.
Hepatits B Virus
The hepatitis B virus (HBV) belongs to the Hepadnaviridae family and shares unique characteristics resembling retroviruses. It is a small DNA virus and serves as the prototype for this viral family. HBV is not limited to humans; related viruses can also be found in various animal species such as woodchucks, ground squirrels, tree squirrels, Peking ducks, and herons. Classifying HBV based on sequence comparison reveals eight distinct genotypes, labeled A to H, each with its own geographic distribution.
When examining infectious serum under an electron microscope, three types of viral particles become visible. Among them, two smaller spherical structures with variable-length filaments can be observed. These structures consist of hepatitis B surface antigen (HBsAg) and host-derived lipids but lack viral nucleic acids, rendering them noninfectious. On the other hand, the infectious HBV virion, known as the Dane particle, displays a double-shelled, spherical structure with a diameter of 42 nm. This virion comprises a lipid envelope containing HBsAg, encasing an inner nucleocapsid. The nucleocapsid consists of hepatitis B core antigen (HBcAg) complexed with virally encoded polymerase and the viral DNA genome.
Hepatits B Virus Life Cycle
The HBV life cycle involves several stages. Upon entering the bloodstream, the virus attaches to specific receptors on hepatocytes, the liver cells. It then enters the cells and releases its viral DNA into the nucleus, where it is converted into a covalently closed circular DNA (cccDNA) form. The cccDNA serves as a template for the transcription of viral RNA, which is then utilized for the synthesis of viral proteins and replication of the viral genome. New viral particles are assembled and released from infected hepatocytes, leading to the spread of the infection. The HBV life cycle is complex and tightly regulated, involving various viral and host factors, and understanding its intricacies is crucial for the development of effective antiviral therapies.
Hepatits B Virus X-Interacting Protein
HBV X-Interacting protein (HBXIP) is a crucial component involved in the pathogenesis of Hepatitis B virus (HBV) infection. HBx is a 154-amino acid (aa) protein that facilitates the efficient replication of HBV by stimulating HBV gene expression from the cccDNA template. However, the mechanism by which HBx interacts with host proteins and facilitates HBV replication is not precisely known. Additionally, HBXIP has been implicated in promoting cellular transformation, tumorigenesis, and progression of liver cancer associated with HBV infection. Studies on HBXIP are crucial for understanding the molecular mechanisms underlying HBV pathogenesis and may provide valuable insights into the development of novel therapeutic strategies targeting this interaction to combat HBV-related liver diseases
HBXIP ELISA Kit FAQs
What is the HBXIP ELISA kit used for?
The HBXIP ELISA kit is specifically designed for the detection and quantification of Hepatitis B virus X-interacting protein (HBXIP) in biological samples. HBXIP is a protein that interacts with the Hepatitis B virus X protein (HBx), a regulatory protein produced by the hepatitis B virus. The HBXIP ELISA kit enables researchers and healthcare professionals to measure the levels of HBXIP, which can provide valuable insights into various biological processes and disease conditions.
What are the advantages of using the HBXIP ELISA Kit?
The HBXIP ELISA Kit offers several advantages, including high sensitivity, accuracy, and reproducibility. It provides a user-friendly and reliable method to quantify HbsAg levels in biological specimens, allowing for precise measurements and robust data analysis.
What sample types are compatible with the HBXIP ELISA Kit?
The HBXIP 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 HBXIP levels in different biological matrices.
What are the storage requirements for the HBXIP ELISA Kit?
The HBXIP 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 HBXIP ELISA Kit.
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