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Mouse CD68 / Macrosialin ELISA Kit

SKU:
MOFI00564
Product Type:
ELISA Kit
Size:
96 Assays
Uniprot:
P31996
Sensitivity:
0.469ng/ml
Range:
0.781-50ng/ml
ELISA Type:
Sandwich
Synonyms:
CD68, Gp110, LAMP4, SCARD1, SCARD1, SR-D1, CD68 antigenmacrophage antigen CD68, molecule, macrosialin, SCARD1, scavenger receptor class D, member 1
Reactivity:
Mouse
€649
Frequently bought together:

Description

Mouse CD68 / Macrosialin ELISA Kit

CD68, also known as macrosialin, is a glycoprotein primarily expressed in cells of the monocyte/macrophage lineage. It is a transmembrane protein found in the lysosomes of these cells and is involved in various cellular processes. The Mouse Macrosialin ELISA Kit is designed to quantitatively measure the concentration of mouse macrosialin (CD68) in biological samples. It allows researchers to study the levels of this glycoprotein in various tissues and cell types, providing valuable insights into macrophage activation, immune responses, and inflammatory processes.

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:

Mouse CD68 / Macrosialin ELISA Kit

Product Code:

MOFI00564

Size:

96 Assays

Alias:

CD68, Gp110, LAMP4, SCARD1, SCARD1, SR-D1, CD68 antigenmacrophage antigen CD68, molecule, macrosialin, SCARD1, scavenger receptor class D, member 1

Detection Method:

Sandwich ELISA

Application:

This immunoassay kit allows for the in vitro quantitative determination of Mouse CD68 concentrations in serum plasma and other biological fluids.

Sensitivity:

0.469ng/ml

Range:

0.781-50ng/ml

Storage:

4°C for 6 months

Note:

For Research Use Only

Additional Information

Recovery

Matrices listed below were spiked with certain level of Mouse CD68 and the recovery rates were calculated by comparing the measured value to the expected amount of Mouse CD68 in samples.

Matrix

Recovery Range (%)

Average (%)

serum (n=5)

86-99

95

EDTA plasma (n=5)

88-100

93

UFH plasma (n=5)

85-99

91

Linearity:

The linearity of the kit was assayed by testing samples spiked with appropriate concentration of Mouse CD68 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%

87-103%

87-97%

EDTA plasma (n=5)

85-97%

93-101%

8-95%

UFH Plasma (n=5)

80-99%

80-99%

82-100%

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

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 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.

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.

CD68 Background

Cd68 marker / Macrosialin

CD68, also known as macrosialin, is a glycoprotein primarily expressed in cells of the monocyte/macrophage lineage. It is a transmembrane protein found in the lysosomes of these cells and is involved in various cellular processes. CD68 serves as a well-established marker for identifying and studying macrophages in tissues and plays a crucial role in the immune response.

CD68 Gene

In mice, the CD68 gene is located on chromosome 11 and contains 6 exons. It is also known as Emr1 (EGF-like module-containing mucin-like hormone receptor-like 1).

CD68 Expression

CD68 is predominantly expressed in cells of the monocyte/macrophage lineage, including monocytes, tissue-resident macrophages, and activated macrophages. Its expression is induced during monocyte differentiation into macrophages. Additionally, it can be detected in other phagocytic cells, such as dendritic cells and osteoclasts.

CD68 Function

The primary function of CD68 is associated with the process of endocytosis and phagocytosis. It is involved in the internalization and degradation of extracellular material, such as microorganisms, cellular debris, and macromolecules. As a lysosomal protein, CD68 participates in the breakdown of engulfed material, leading to antigen presentation and activation of the immune response.

Moreover, CD68 is considered a scavenger receptor, playing a role in host defense mechanisms and tissue homeostasis. By recognizing and binding to various ligands, CD68 aids in the clearance of potentially harmful substances and contributes to the immune system's ability to control infections and regulate inflammation.

CD68 Clinical Significance

CD68 has significant clinical importance due to its role as a marker for macrophages in tissues. Immunohistochemical staining for CD68 is commonly used in pathology and research to identify and quantify macrophage infiltration in various diseases, including infectious diseases, autoimmune disorders, and cancer.

The expression of CD68 in specific disease states can provide valuable diagnostic and prognostic information. For instance, increased CD68 expression in tumor-associated macrophages has been linked to tumor progression and poorer outcomes in certain types of cancer. In contrast, in infectious diseases, such as tuberculosis, CD68 expression can help identify the presence of mycobacteria in tissues and facilitate diagnosis.

Mouse CD68 ELISA Kit FAQs

Q: What is the purpose of the Mouse Macrosialin ELISA Kit?

The Mouse Macrosialin ELISA Kit is designed to quantitatively measure the concentration of mouse macrosialin (CD68) in biological samples. It allows researchers to study the levels of this glycoprotein in various tissues and cell types, providing valuable insights into macrophage activation, immune responses, and inflammatory processes.

Q: What type of samples can be used with this ELISA Kit?

This ELISA Kit is optimized for measuring mouse macrosialin in various biological samples, including serum, plasma, cell lysates, and tissue homogenates. However, it is essential to follow the specific sample preparation instructions provided in the kit's protocol.

Q: Is this ELISA Kit specific to mouse macrosialin?

Yes, this ELISA Kit is specifically designed and validated for the quantitative measurement of mouse macrosialin (CD68) in biological samples. It does not cross-react with macrosialin from other species.

Q: Where can I find additional technical support or assistance with the kit?

For any technical inquiries or assistance regarding the kit, you can reach out to our team. They will be available to answer your questions and provide the necessary guidance to ensure a successful experiment.

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