null

Dog C-X-C motif chemokine 10 (CXCL10) ELISA Kit

SKU:
CNEB0082
Product Type:
ELISA Kit
Size:
96 Assays
Uniprot:
Q5KSV9
ELISA Type:
Sandwich
Reactivity:
Canine
$779
Frequently bought together:

Description

Dog C-X-C motif chemokine 10 (CXCL10) ELISA Kit

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:

Dog C-X-C motif chemokine 10 (CXCL10) ELISA Kit

Product Code:

CNEB0082

Alias:

C-X-C motif chemokine 10, Small-inducible cytokine B10, CXCL10

Uniprot:

Q5KSV9

Reactivity:

Dog

Range:

Please contact us for more information

Detection Method:

Sandwich

Size:

96 Assays

Storage:

Please see kit components below for exact storage details

Note:

For Research Use Only

Protein Information

UniProt Protein Function:

Function: Chemotactic for monocytes and T-lymphocytes. Binds to CXCR3 By similarity.Subcellular location: Secreted By similarity. Sequence similarities: Belongs to the intercrine alpha (chemokine CxC) family.

UniProt Code:

Q5KSV9

NCBI GenInfo Identifier:

58219486

NCBI Gene ID:

478432

NCBI Accession:

NP_001010949.1

UniProt Related Accession:

Q5KSV9

Molecular Weight:

10.1kDa

Kit Components

Component Quantity Storage

ELISA Microplate (Dismountable)

8x12 strips

-20°C

Lyophilized Standard

2

-20°C

Sample Diluent

20ml

-20°C

Assay Diluent A

10mL

-20°C

Assay Diluent B

10ml

-20°C

Detection Reagent A

60µL

-20°C

Detection Reagent B

120µL

-20°C

Wash Buffer (25X)

30ml

4°C

Substrate

10mL

4°C

Stop Solution

10ml

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: The below protocol is a sample protocol. Protocols are specific to each batch/lot. For the correct instructions please follow the protocol included in your kit.

Allow all reagents to reach room temperature (Please do not dissolve the reagents at 37°C directly). All the reagents should be mixed thoroughly by gently swirling before pipetting. Avoid foaming. Keep appropriate numbers of strips for 1 experiment and remove extra strips from microtiter plate. Removed strips should be resealed and stored at -20°C until the kits expiry date. Prepare all reagents, working standards and samples as directed in the previous sections. Please predict the concentration before assaying. If values for these are not within the range of the standard curve, users must determine the optimal sample dilutions for their experiments. We recommend running all samples in duplicate.

Step Procedure

1.

Add Sample: Add 100µL of Standard, Blank, or Sample per well. The blank well is added with Sample diluent. Solutions are added to the bottom of micro ELISA plate well, avoid inside wall touching and foaming as possible. Mix it gently. Cover the plate with sealer we provided. Incubate for 120 minutes at 37°C.

2.

Remove the liquid from each well, don't wash. Add 100µL of Detection Reagent A working solution to each well. Cover with the Plate sealer. Gently tap the plate to ensure thorough mixing. Incubate for 1 hour at 37°C. Note: if Detection Reagent A appears cloudy warm to room temperature until solution is uniform.

3.

Aspirate each well and wash, repeating the process three times. Wash by filling each well with Wash Buffer (approximately 400µL) (a squirt bottle, multi-channel pipette,manifold dispenser or automated washer are needed). Complete removal of liquid at each step is essential. After the last wash, completely remove remaining Wash Buffer by aspirating or decanting. Invert the plate and pat it against thick clean absorbent paper.

4.

Add 100µL of Detection Reagent B working solution to each well. Cover with the Plate sealer. Incubate for 60 minutes at 37°C.

5.

Repeat the wash process for five times as conducted in step 3.

6.

Add 90µL of Substrate Solution to each well. Cover with a new Plate sealer and incubate for 10-20 minutes at 37°C. Protect the plate from light. The reaction time can be shortened or extended according to the actual color change, but this should not exceed more than 30 minutes. When apparent gradient appears in standard wells, user should terminatethe reaction.

7.

Add 50µL of Stop Solution to each well. If color change does not appear uniform, gently tap the plate to ensure thorough mixing.

8.

Determine the optical density (OD value) of each well at once, using a micro-plate reader set to 450 nm. User should open the micro-plate reader in advance, preheat the instrument, and set the testing parameters.

9.

After experiment, store all reagents according to the specified storage temperature respectively until their expiry.

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.

CXCL10 Background

C-X-C motif chemokine 10 (CXCL10), also known as interferon gamma-induced protein 10 (IP-10), is a small protein that belongs to the chemokine family. Chemokines are signaling molecules involved in immune responses, inflammation, and the recruitment of immune cells to specific sites in the body.

CXCL10 Production

CXCL10 is primarily produced by various cell types, including immune cells such as monocytes, macrophages, and dendritic cells. It is induced by the pro-inflammatory cytokine interferon gamma (IFN-γ), which is released during immune responses to viral or bacterial infections, as well as in certain autoimmune diseases.

CXCL10 Gene

The CXCL10 gene encodes the CXCL10 protein, which is involved in immune responses and inflammation. Upon activation, the CXCL10 protein binds to its receptor, CXCR3, on immune cells, initiating signaling pathways that regulate cell migration, immune cell activation, and cytokine production. The CXCL10 gene is associated with various diseases, including autoimmune conditions and certain cancers, and studying its variations provides insights into disease susceptibility and potential therapeutic targets.

CXCL10 Pathway

The CXCL10 pathway involves the binding of the CXCL10 protein to its receptor, CXCR3, on immune cells. This binding triggers intracellular signaling pathways, including JAK/STAT, PI3K, and MAPK, leading to the activation of immune cells and the induction of cell migration. CXCL10 acts as a chemoattractant, guiding immune cells to sites of inflammation or infection, and also regulates inflammatory processes by influencing immune cell activation and cytokine production. The CXCL10 pathway is implicated in immune responses, autoimmune diseases, infections, and cancer, making it a target for therapeutic interventions.

CXCL10 Function

The primary function of CXCL10 is to attract and activate immune cells, particularly T cells and natural killer (NK) cells. It acts as a chemoattractant, guiding these cells to sites of inflammation or infection. CXCL10 binds to its specific receptor, called CXCR3, which is expressed on the surface of immune cells. This interaction triggers signaling pathways that promote cell migration and activation. CXCL10 has been implicated in various physiological and pathological processes. It plays a crucial role in antiviral immune responses by attracting immune cells to infected tissues.

CXCL10 FAQs

What is the CXCL10 ELISA Kit?

The CXCL10 ELISA Kit is a specialized assay kit designed to measure the levels of CXCL10 (C-X-C motif chemokine 10) in biological samples.

What are the advantages of using the CXCL10 ELISA Kit?

The CXCL10 ELISA Kit provides a sensitive and specific method for accurately measuring CXCL10 levels in various biological samples. he kit's quantitative results allow for precise comparisons between different samples or experimental conditions. The Kit offers convenience with its ready-to-use components and straightforward protocol, saving time and effort in sample analysis.

Where can I find more information about the CXCL10 ELISA Kit?

For more detailed information about the CXCL10 ELISA Kit, 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.

Related Products