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Human CD163 ELISA Kit

SKU:
HUFI00068
Product Type:
ELISA Kit
Size:
96 Assays
Uniprot:
Q86VB7
Sensitivity:
0.938ng/ml
Range:
1.563-100ng/ml
ELISA Type:
Sandwich
Synonyms:
sCD163, Soluble Cluster of Differentiation 163, M130, MM130
Reactivity:
Human
€499
Frequently bought together:

Description

Human CD163 ELISA Kit

Cluster of Differentiation 163 (CD163) is a protein found on the surface of natural killer cells, which play an important role in fighting cancer cells and infected cells. CD163 also functions in the activation and proliferation of T cells. CD163 is used as a marker for the differentiation of Th1 cells. The Assay Genie Human CD163 ELISA kit is a highly sensitive assay for the quantitative measurement of CD163 in serum, blood, plasma, cell culture supernatant, and tissue samples.

system_update_alt Datasheet system_update_alt MSDS

Key Features

Save Time Pre-coated 96 well plates
Quick Start Kit includes all necessary reagents
Publication Ready Reproducible and Reliable results

Overview

Product Name:

Human CD163 ELISA Kit

Product Code:

HUFI00068

Size:

96 Assays

Alias:

sCD163, Soluble Cluster of Differentiation 163, M130, MM130

Detection Method:

Sandwich ELISA, Double Antibody

Application:

This immunoassay kit allows for the in vitro quantitative determination of Human sCD163 concentrations in serum plasma and other biological fluids.

Sensitivity:

0.938ng/ml

Range:

1.563-100ng/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 sCD163 and the recovery rates were calculated by comparing the measured value to the expected amount of Human sCD163 in samples.

Matrix

Recovery Range (%)

Average (%)

serum(n=5)

85-103

96

EDTA plasma(n=5)

85-105

96

UFH plasma(n=5)

88-104

96

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.

Sample

1:2

1:4

1:8

serum(n=5)

88-104%

85-97%

89-100%

EDTA plasma(n=5)

82-96%

90-97%

86-99%

UFH plasma(n=5)

86-98%

86-100%

82-100%

CV(%)

Intra-Assay: CV<8%
Inter-Assay: CV<10%

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

Heading 1 Heading 2

UniProt

NCBI GenInfo Identifier

NCBI Gene ID

NCBI Official Symbol

CD163

Protein Family

Scavenger receptor cysteine-rich type 1 protein

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.

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.

CD163 Background

CD163 Background

CD163 is a cell surface receptor protein primarily expressed on monocytes and macrophages, particularly those associated with tissues involved in immune responses and inflammation. This receptor is a transmembrane glycoprotein that plays a key role in modulating the immune system's response to various stimuli. The extracellular portion of CD163 can also be shed from the cell surface, generating a soluble form known as sCD163. This soluble form has been implicated in several physiological and pathological conditions, making it a valuable biomarker for certain diseases.

CD163 Function

CD163 serves as a multifunctional receptor involved in several essential processes within the immune system. Its primary role is to facilitate the clearance of hemoglobin-haptoglobin complexes from circulation. When red blood cells break down, hemoglobin is released and can be toxic if not appropriately managed. CD163 binds to the haptoglobin-hemoglobin complex, leading to its internalization and degradation within macrophages. This mechanism helps prevent the accumulation of free hemoglobin and its potential harm to tissues. Moreover, CD163 has been associated with anti-inflammatory actions; its expression can be upregulated in response to anti-inflammatory stimuli, indicating its involvement in dampening inflammatory responses. The release of sCD163 into the bloodstream might also have immunomodulatory effects. Due to its diverse roles, CD163 and sCD163 have garnered interest as potential targets and biomarkers in various inflammatory and hematological disorders.

CD163 ELISA Kit FAQs

What is the Human CD163 ELISA Kit used for?

The Human CD163 ELISA kit is a diagnostic tool commonly used in research and clinical settings to measure the levels of CD163, a cell surface receptor primarily found on macrophages and monocytes. CD163 is known to play a crucial role in modulating the immune response and has been implicated in various inflammatory and immune-related conditions. By quantifying CD163 levels in biological samples like blood serum or plasma, the ELISA kit helps researchers and clinicians gain insights into the activation status of macrophages, monitor immune system activity, and potentially assess the severity of certain diseases such as autoimmune disorders, infections, and inflammatory diseases.

What are the advantages of using the Human CD163 ELISA Kit?

The Human CD163 ELISA Kit offers several advantages, including high sensitivity, accuracy, and reproducibility. It provides a user-friendly and reliable method to quantify CD163 levels in biological specimens, allowing for precise measurements and robust data analysis.

What sample types are compatible with Human CD163 ELISA Kit?

The Human CD163 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 CD163 levels in different biological matrices.

What are the storage requirements with Human CD163 ELISA Kit?

The Human CD163 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 Human CD163 ELISA Kit.