Canine KIM-1 ELISA Kit (CNFI00023)- High Sensitivity

Key Features

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

Overview


Product Name:	Canine KIM-1(Kidney Injury Molecule 1) ELISA Kit
Product Code:	CNFI00023
Size:	96 Assays
Alias:	KIM-1 ELISA Kit, HAVCR1 ELISA Kit, HAVCR ELISA Kit
Detection Method:	Sandwich, Double Antibody
Sensitivity:	0.188ng/ml
Range:	0.313-20ng/ml
Storage:	2-8°C for 6 months
Note:	For Research Use Only

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.

Canine KIM-1 Background

Kidney Injury Molecule 1 (KIM-1) is a protein that is primarily expressed in the kidney, although it can also be found in other tissues. It is considered a biomarker of kidney injury and is used to detect and monitor kidney damage.

KIM-1 Structure and Expression

KIM-1, also known as T-cell immunoglobulin mucin domain 1 (TIM-1), belongs to the TIM family of proteins. It is a type I transmembrane glycoprotein consisting of an extracellular region, a transmembrane domain, and an intracellular domain. The extracellular region of KIM-1 contains an immunoglobulin variable-like (IgV) domain and a mucin-like domain. KIM-1 expression is not limited to the kidney; it has also been detected in other tissues, such as the liver, lung, and testes, although at lower levels.

Role of KIM-1 in Kidney Injury and Repair

When the kidneys are injured, KIM-1 is upregulated and released into urine or blood samples, providing valuable information about the extent of kidney injury and disease progression. It is useful in diagnosing and monitoring conditions such as acute kidney injury (AKI), chronic kidney disease (CKD), and renal transplant rejection. Additionally, KIM-1 plays a role in the repair and regeneration of kidney cells, aiding in the clearance of dead cells and debris from the kidney tubules. However, sustained elevation of KIM-1 levels suggests persistent or severe kidney injury, indicating ongoing damage and impaired kidney function.

Canine KIM-1 FAQs

What is the Canine KIM-1 ELISA Kit?

The Canine KIM-1 ELISA Kit is a scientific tool designed for the quantitative measurement of Canine KIM-1 (Kidney Injury Molecule 1) levels in biological samples.

What are the benefits of using the Canine KIM-1 ELISA Kit?

The Canine KIM-1 ELISA Kit provides several benefits for researchers and professionals working in veterinary or biomedical fields. It offers a reliable and sensitive method to assess kidney injury in canines, aiding in the diagnosis and monitoring of kidney diseases. By quantifying Canine KIM-1 levels, the kit facilitates early detection, treatment evaluation, and research investigations related to canine kidney health.

Where can I find more information about the Canine KIM-1 ELISA Kit?

For more detailed information about the Canine KIM-1 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.