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Human F13A / Factor XIII ELISA Kit

SKU:
HUFI00762
Product Type:
ELISA Kit
Size:
96 Assays
Uniprot:
P00488
Sensitivity:
0.094ng/ml
Range:
0.156-10ng/ml
ELISA Type:
Sandwich
Synonyms:
F13A1, F13A,, Coagulation Factor XIII A1 Polypeptide, Transglutaminase A chain, Coagulation factor XIIIa
Reactivity:
Human
Research Area:
Cardiovascular
€599
Frequently bought together:

Description

Human F13A/Factor XIII ELISA Kit

The Human Factor XIII ELISA Kit is a highly reliable and accurate tool for the detection of Factor XIII levels in human serum, plasma, and cell culture supernatants. This kit offers high sensitivity and specificity, allowing for precise and reproducible results in various research applications.Factor XIII is a key protein involved in the final stages of the blood clotting process, playing a crucial role in maintaining hemostasis and preventing excessive bleeding. Dysregulation of Factor XIII activity has been linked to various bleeding disorders and thrombotic events, making it a valuable biomarker for studying these conditions and developing new treatment strategies.

With its user-friendly design and comprehensive instructions, the Human Factor XIII ELISA Kit is an essential tool for researchers and clinicians working in the fields of hematology, vascular biology, and coagulation disorders. Trust in this kit to deliver accurate results and advance your research efforts in understanding the role of Factor XIII in health and disease.

Product Name:Human F13A / Factor XIII ELISA Kit
Product Code:HUFI00762
Size:96 Assays
Alias:F13A1, F13A, Coagulation Factor XIII A1 Polypeptide, Transglutaminase A chain, Coagulation factor XIIIa
Detection method:Sandwich ELISA, Double Antibody
Application:This immunoassay kit allows for the in vitro quantitative determination of Human F13A1 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
Recovery:Matrices listed below were spiked with certain level of Human F13A1 and the recovery rates were calculated by comparing the measured value to the expected amount of Human F13A1 in samples.
MatrixRecovery range(%)Average(%)
serum(n=5)92-10296
EDTA plasma(n=5)86-10595
UFH plasma(n=5)86-10194
Linearity:The linearity of the kit was assayed by testing samples spiked with appropriate concentration of Human F13A1 and their serial dilutions. The results were demonstrated by the percentage of calculated concentration to the expected.
Sample1:21:41:8
serum(n=5)85-100%85-105%89-102%
EDTA plasma(n=5)84-101%86-96%85-101%
UFH plasma(n=5)80-98%81-92%87-96%
CV(%):Intra-Assay: CV<8%
Inter-Assay: CV<10%
ComponentQuantityStorage
ELISA Microplate (Dismountable)8×12 strips4°C for 6 months
Lyophilized Standard24°C/-20°C
Sample/Standard Dilution Buffer20ml4°C
Biotin-labeled Antibody(Concentrated)120ul4°C (Protect from light)
Antibody Dilution Buffer10ml4°C
HRP-Streptavidin Conjugate(SABC)120ul4°C (Protect from light)
SABC Dilution Buffer10ml4°C
TMB Substrate10ml4°C (Protect from light)
Stop Solution10ml4°C
Wash Buffer(25X)30ml4°C
Plate Sealer5-

Other materials and equipment 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
UniprotP00488
UniProt Protein Function:F13A1: Factor XIII is activated by thrombin and calcium ion to a transglutaminase that catalyzes the formation of gamma-glutamyl- epsilon-lysine cross-links between fibrin chains, thus stabilizing the fibrin clot. Also cross-link alpha-2-plasmin inhibitor, or fibronectin, to the alpha chains of fibrin. Defects in F13A1 are the cause of factor XIII subunit A deficiency (FA13AD). FA13AD is an autosomal recessive disorder characterized by a life-long bleeding tendency, impaired wound healing and spontaneous abortion in affected women. Belongs to the transglutaminase superfamily. Transglutaminase family.
UniProt Protein Details:Protein type:Transferase; EC 2.3.2.13

Chromosomal Location of Human Ortholog: 6p25.3-p24.3

Cellular Component: extracellular region

Molecular Function:protein-glutamine gamma-glutamyltransferase activity; metal ion binding

Biological Process: platelet activation; platelet degranulation; peptide cross-linking; blood coagulation

Disease: Thrombophilia Due To Thrombin Defect; Myocardial Infarction, Susceptibility To; Factor Xiii, A Subunit, Deficiency Of

NCBI Summary:This gene encodes the coagulation factor XIII A subunit. Coagulation factor XIII is the last zymogen to become activated in the blood coagulation cascade. Plasma factor XIII is a heterotetramer composed of 2 A subunits and 2 B subunits. The A subunits have catalytic function, and the B subunits do not have enzymatic activity and may serve as plasma carrier molecules. Platelet factor XIII is comprised only of 2 A subunits, which are identical to those of plasma origin. Upon cleavage of the activation peptide by thrombin and in the presence of calcium ion, the plasma factor XIII dissociates its B subunits and yields the same active enzyme, factor XIIIa, as platelet factor XIII. This enzyme acts as a transglutaminase to catalyze the formation of gamma-glutamyl-epsilon-lysine crosslinking between fibrin molecules, thus stabilizing the fibrin clot. It also crosslinks alpha-2-plasmin inhibitor, or fibronectin, to the alpha chains of fibrin. Factor XIII deficiency is classified into two categories: type I deficiency, characterized by the lack of both the A and B subunits; and type II deficiency, characterized by the lack of the A subunit alone. These defects can result in a lifelong bleeding tendency, defective wound healing, and habitual abortion. [provided by RefSeq, Jul 2008]
UniProt Code:P00488
NCBI GenInfo Identifier:119395709
NCBI Gene ID:2162
NCBI Accession:NP_000120.2
UniProt Secondary Accession:P00488,Q59HA7, Q8N6X2, Q96P24, Q9BX29,
UniProt Related Accession:P00488
Molecular Weight:312,000
NCBI Full Name:coagulation factor XIII A chain
NCBI Synonym Full Names:coagulation factor XIII, A1 polypeptide
NCBI Official Symbol:F13A1
NCBI Official Synonym Symbols:F13A
NCBI Protein Information:coagulation factor XIII A chain; TGase; factor XIIIa; fibrinoligase; FSF, A subunit; coagulation factor XIIIa; transglutaminase A chain; transglutaminase. plasma; fibrin stabilizing factor, A subunit; coagulation factor XIII, A polypeptide; protein-glutamine gamma-glutamyltransferase A chain; bA525O21.1 (coagulation factor XIII, A1 polypeptide)
UniProt Protein Name:Coagulation factor XIII A chain
UniProt Synonym Protein Names:Protein-glutamine gamma-glutamyltransferase A chain; Transglutaminase A chain
Protein Family:36 kDa major membrane protein
UniProt Gene Name:F13A1
UniProt Entry Name:F13A_HUMAN

*Note: Protocols are specific to each batch/lot. For the correct 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.

StepProtocol
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.

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 TypeProtocol
SerumIf 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.

PlasmaCollect 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 FluidCollect 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 supernatantCollect 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 lysatesSolubilize 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 homogenatesThe 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 lysatesRinse 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 MilkCollect 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.

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