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Human Syndecan-1 ELISA Kit

SKU:
HUFI00239
Product Type:
ELISA Kit
Size:
96 Assays
Uniprot:
P18827
Sensitivity:
0.094ng/ml
Range:
0.156-10ng/ml
ELISA Type:
Sandwich
Synonyms:
Syndecan?1, SDC1, CD138, SDC, syndecan, syndecan 1, syndecan proteoglycan 1
Reactivity:
Human
€599
Frequently bought together:

Description

Human Syndecan-1 ELISA Kit

Syndecan-1, also known as CD138, is a cell surface proteoglycan with a single transmembrane domain. It is widely expressed on epithelial cells and plasma cells. Syndecan-1 functions as a pivotal regulator of cell-matrix interactions, mediating adhesion, signaling, and extracellular matrix organization through its heparan sulfate chains and interactions with various ligands.

system_update_alt Datasheet system_update_alt MSDS

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 Syndecan-1 ELISA Kit

Product Code:

HUFI00239

Size:

96 Assays

Alias:

SDC1, CD138, SDC, syndecan, syndecan 1, syndecan proteoglycan 1

Detection Method:

Sandwich ELISA, Double Antibody

Application:

This immunoassay kit allows for the in vitro quantitative determination of Human SDC1 concentrations in serum plasma and other biological fluids.

Sensitivity:

0.094ng/ml

Range:

0.156-10ng/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 SDC1 and the recovery rates were calculated by comparing the measured value to the expected amount of Human SDC1 in samples.

Heading 1 Heading 2

UniProt

NCBI GenInfo Identifier

NCBI Gene ID

Molecular Weight

Protein Family

Heading 1 Heading 2

UniProt

NCBI GenInfo Identifier

NCBI Gene ID

Molecular Weight

Protein Family

Matrix

Recovery Range (%)

Average (%)

serum(n=5)

94-105

100

EDTA plasma(n=5)

90-97

94

UFH plasma(n=5)

87-101

95

Linearity

The linearity of the kit was assayed by testing samples spiked with appropriate concentration of Human SDC1 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)

91-105%

85-100%

88-102%

EDTA plasma(n=5)

86-98%

83-92%

91-101%

UFH plasma(n=5)

82-100%

81-96%

84-91%

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

Molecular Weight

32,462 Da

Protein Family

COMPASS component

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.

Syndecan-1 Background

Syndecan-1 Background

Syndecan-1, also known as CD138, is a cell surface proteoglycan that belongs to the syndecan family of transmembrane proteins. It is primarily found on the surfaces of epithelial cells, plasma cells, and other cell types in various tissues. Syndecan-1 serves as a multifunctional molecule involved in cell adhesion, signaling, and modulation of the extracellular matrix. In the context of plasma cells, CD138 is commonly used as a marker for identifying and isolating these cells, especially in the study of hematopoietic malignancies and multiple myeloma.

Syndecan-1 Structure and Function

Syndecan-1 consists of a short cytoplasmic domain, a single transmembrane domain, and an extracellular domain that contains heparan sulfate chains attached to a core protein. These heparan sulfate chains facilitate interactions with various ligands, including growth factors, cytokines, and components of the extracellular matrix. Through these interactions, syndecan-1 plays a crucial role in cell adhesion, migration, and signal transduction. Additionally, syndecan-1 can modulate cell-matrix interactions by participating in the assembly of focal adhesions and influencing the binding of integrins to the extracellular matrix. Its presence on the surface of plasma cells also contributes to the structural integrity of the cells and their interactions with bone marrow stromal cells.

Syndecan-1 ELISA Kit FAQs

What is the Syndecan-1 ELISA Kit used for?

The syndecan-1 ELISA kit is used for the quantitative measurement of syndecan-1 (CD138) levels in biological samples such as serum, plasma, or cell culture supernatants. This ELISA Kit aids researchers and clinicians in studying various physiological and pathological conditions, including hematological malignancies like multiple myeloma, as well as other inflammatory and tissue remodeling processes where syndecan-1 shedding and expression are relevant biomarkers.

What are the advantages of using the Syndecan-1 ELISA Kit?

The Syndecan-1 ELISA Kit offers several advantages, including high sensitivity, accuracy, and reproducibility. It provides a user-friendly and reliable method to quantify Syndecan-1 levels in biological specimens, allowing for precise measurements and robust data analysis.

What sample types are compatible with Syndecan-1 ELISA Kit?

The Syndecan-1 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 Syndecan-1 levels in different biological matrices.

What are the storage requirements with Syndecan-1 ELISA Kit?

The Syndecan-1 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 Syndecan-1 ELISA Kit.