CD13 ELISA Kit

Catalog No. ABIN1889386

Product Details CD13 ELISA Kit

Target: CD13 (ANPEP) (Alanyl (Membrane) Aminopeptidase (ANPEP))

Binding Specificity: AA 69-966

Reactivity: Mouse

Detection Method: Colorimetric

Method Type: Sandwich ELISA

Detection Range: 312-20.000 pg/mL

Minimum Detection Limit: 312 pg/mL

Application: ELISA

Purpose: Sandwich High Sensitivity ELISA kit for Quantitative Detection of Mouse CD13/Aminopeptidase N

Brand: PicoKine™

Sample Type: Cell Culture Supernatant, Serum, Plasma (heparin), Plasma (EDTA)

Analytical Method: Quantitative

Specificity: Expression system for standard: NSO
Immunogen sequence: K69-S966

Cross-Reactivity (Details): There is no detectable cross-reactivity with other relevant proteins.

Sensitivity: <10pg/mL

Material not included: Microplate reader in standard size. Automated plate washer. Adjustable pipettes and pipette tips. Multichannel pipettes are recommended in the condition of large amount of samples in the detection. Clean tubes and Eppendorf tubes. Washing buffer (neutral PBS or TBS). Preparation of 0.01M TBS: Add 1.2g Tris, 8.5g Nacl

Immunogen: Expression system for standard: NSO
Immunogen sequence: K69-S966

Application Details

Application Notes: Before using Kit, spin tubes and bring down all components to bottom of tube. Duplicate well assay was recommended for both standard and sample testing.

Comment:

Tissue Specificity: Highly expressed in intestinal tract and kidney, present in liver, lymph node, spleen, and brain. Found as well in monocytes, macrophages, dendritic cells, veiled cells and B-cells but not on T-cells and thymocytes. .

Plate: Pre-coated

Protocol: mouse CD13 ELISA Kit was based on standard sandwich enzyme-linked immune-sorbent assay technology. A monoclonal antibody from rat specific for CD13 has been precoated onto 96-well plates. Standards(NSO, K69-S966) and test samples are added to the wells, a biotinylated detection polyclonal antibody from goat specific for CD13 is added subsequently and then followed by washing with PBS or TBS buffer. Avidin-Biotin-Peroxidase Complex was added and unbound conjugates were washed away with PBS or TBS buffer. HRP substrate TMB was used to visualize HRP enzymatic reaction. TMB was catalyzed by HRP to produce a blue color product that changed into yellow after adding acidic stop solution. The density of yellow is proportional to the mouse CD13 amount of sample captured in plate.

Assay Procedure:

Aliquot 0.1 mL per well of the 20,000pg/mL, 10,000pg/mL, 5000pg/mL, 2500pg/mL, 1250pg/mL, 625pg/mL, 312pg/mL mouse CD13 standard solutions into the precoated 96-well plate. Add 0.1 mL of the sample diluent buffer into the control well (Zero well). Add 0.1 mL of each properly diluted sample of mouse cell culture supernates, serum or plasma(heparin, EDTA) to each empty well. See "Sample Dilution Guideline" above for details. It is recommended that each mouse CD13 standard solution and each sample be measured in duplicate.

Assay Precision:

• Sample 1: n=16, Mean(ng/ml): 3.15, Standard deviation: 0.164, CV(%): 5.2
• Sample 2: n=16, Mean(ng/ml): 6.64, Standard deviation: 0.372, CV(%): 5.6
• Sample 3: n=16, Mean(ng/ml): 13.09, Standard deviation: 0.785, CV(%): 6.0,
• Sample 1: n=24, Mean(ng/ml): 3.36, Standard deviation: 0.218, CV(%): 6.5
• Sample 2: n=24, Mean(ng/ml): 6.97, Standard deviation: 0.474, CV(%): 6.8
• Sample 3: n=24, Mean(ng/ml): 14.15, Standard deviation: 1.02, CV(%): 7.2

Restrictions: For Research Use only

Handling

Handling Advice: Avoid multiple freeze-thaw cycles.

Storage: -20 °C,4 °C

Storage Comment: Store at 4°C for 6 months, at -20°C for 12 months. Avoid multiple freeze-thaw cycles

Expiry Date: 12 months

Target Details for CD13

Target: CD13 (ANPEP) (Alanyl (Membrane) Aminopeptidase (ANPEP))

Alternative Name: ANPEP (ANPEP Products)

Background:

Protein Function: Broad specificity aminopeptidase. Plays a role in the final digestion of peptides generated from hydrolysis of proteins by gastric and pancreatic proteases. May be involved in the metabolism of regulatory peptides of diverse cell types, responsible for the processing of peptide hormones, such as angiotensin III and IV, neuropeptides, and chemokines. May have a role in angiogenesis (By similarity). Found to cleave antigen peptides bound to major histocompatibility complex class II molecules of presenting cells. .

Background: Alanine aminopeptidase, also known as ANPEP or CD13. is an enzyme that is used as a biomarker to detect damage to the kidneys, and that may be used to help diagnose certain kidney disorders. It is mapped to 15q26.1. Aminopeptidase N is located in the small-intestinal and renal microvillar membrane, and also in other plasma membranes. In the small intestine, Aminopeptidase N plays a role in the final digestion of peptides generated from hydrolysis of proteins by gastric and pancreatic proteases, and it is also thought to be involved in the metabolism of regulatory peptides by diverse cell types, including small intestinal and renal tubular epithelial cells, macrophages, granulocytes, and synaptic membranes from the CNS. Petrovic et al showed that CD13 was required for endothelial cell invasion in response to bradykinin. Inhibition of CD13 abrogated internalization of bradykinin receptor B2 and reduced endothelial cell motility.

Synonyms: Aminopeptidase N,AP-N,mAPN,3.4.11.2,Alanyl aminopeptidase,Aminopeptidase M,AP-M,Membrane protein p161,Microsomal aminopeptidase,CD13,Anpep,Lap-1, Lap1,

Full Gene Name: Aminopeptidase N

Cellular Localisation: Membrane, Single-pass type II membrane protein.

Gene ID: 16790

UniProt: P97449

Pathways: Peptide Hormone Metabolism, Regulation of Systemic Arterial Blood Pressure by Hormones