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Human MAP1LC3A ELISA Kit

SKU:
HUFI01900
Product Type:
ELISA Kit
Size:
96 Assays
Uniprot:
Q9H492
Sensitivity:
4.688pg/ml
Range:
7.813-500pg/ml
ELISA Type:
Sandwich
Synonyms:
MAP1LC3A, Apg8, APG8a, Apg8p3, ATG8E, MAP1ALC3, MLP3A, ATG8E, Autophagy-related protein LC3 A, Autophagy-related ubiquitin-like modifier LC3 A, LC3, LC3A, lc3-i, ii, MAP1A, 1B light chain 3 A, MAP1ALC3MAP1A, MAP1B LC3 A, MAP1BLC3MAP1A, MAP1B light ch
Reactivity:
Human
Research Area:
Autophagy
€599
Frequently bought together:

Description

Human MAP1LC3A ELISA Kit

The Human MAP1LC3A ELISA Kit is a reliable and accurate tool for measuring levels of the MAP1LC3A protein in human samples such as serum, plasma, and cell culture supernatants. This kit offers high sensitivity and specificity, ensuring precise and reproducible results for a variety of research applications.MAP1LC3A is a key protein involved in autophagy, a cellular process essential for maintaining cell homeostasis and clearing damaged organelles. Dysregulation of autophagy has been implicated in various diseases, including cancer, neurodegenerative disorders, and inflammatory conditions.

By measuring MAP1LC3A levels, researchers can better understand the role of autophagy in disease pathology and potentially identify new therapeutic targets.The Human MAP1LC3A ELISA Kit is a valuable tool for researchers studying autophagy and its implications in disease processes. Its high performance and ease of use make it an essential addition to any laboratory conducting research in this field.

Product Name:Human MAP1LC3A ELISA Kit
Product Code:HUFI01900
Size:96 Assays
Alias:MAP1LC3A, Apg8, APG8a, Apg8p3, ATG8E, MAP1ALC3, MLP3A, ATG8E, Autophagy-related protein LC3 A, Autophagy-related ubiquitin-like modifier LC3 A, LC3, LC3A, lc3-i, ii, MAP1A, 1B light chain 3 A, MAP1ALC3MAP1A, MAP1B LC3 A, MAP1BLC3MAP1A, MAP1B light chain 3 A, microtubule-associated protein 1 light chain 3 alphaMAP1 light chain 3-like protein 1, microtubule-associated proteins 1A, 1B light chain 3, microtubule-associated proteins 1A, 1B light chain 3A
Detection method:Sandwich ELISA, Double Antibody
Application:This immunoassay kit allows for the in vitro quantitative determination of Human MAP1LC3A concentrations in serum plasma and other biological fluids.
Sensitivity:4.688pg/ml
Range:7.813-500pg/ml
Storage:4°C for 6 months
Note:For Research Use Only
Recovery:Matrices listed below were spiked with certain level of Human MAP1LC3A and the recovery rates were calculated by comparing the measured value to the expected amount of Human MAP1LC3A in samples.
MatrixRecovery range(%)Average(%)
serum(n=5)90-10596
EDTA plasma(n=5)85-10596
UFH plasma(n=5)85-10597
Linearity:The linearity of the kit was assayed by testing samples spiked with appropriate concentration of Human MAP1LC3A and their serial dilutions. The results were demonstrated by the percentage of calculated concentration to the expected.
Sample1:21:41:8
serum(n=5)85-103%89-104%89-105%
EDTA plasma(n=5)84-100%82-99%86-96%
UFH plasma(n=5)80-91%85-95%82-94%
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
UniprotQ9H492
UniProt Protein Function:LC3A: is a ubiquitin-like protein that is a constituent of the ATG8-conjugation system, one of two evolutionarily conserved phosphatidylethanolamine conjugation systems necessary for the formation of the autophagosome. The human ATG8 system includes seven ubiquitin-like light chain proteins (LCPs) that are homologs of yeast LC3: MAP1LC3A, -B, -C, GABARAP, GABARAPL1, -2, and -3. Pro-LCPs are cleaved by ATG4B to expose a C-terminal glycine residue, the cytosolic LCP-I form. The exposed C-terminus is conjugated to the head group amine of phosphatidylethanolamine (PE) through an amide bond by a sequence of ubiquitination-like reactions that involves an E1 (ATG7), an E2 (ATG3), and an E3 (a complex including ATG5, ATG12, and ATG16L). The PE-congugated form (LCP-II) is tightly associated with the autophagosomal membrane. The LCP-II forms can also be delipidated by the ATG4 proteases: most of the LCPs are delipidated and liberated from the membrane before autophagosomes fuse with lysosomes. Two isoforms of the human protein are produced by alternative splicing.
UniProt Protein Details:Protein type:Microtubule-binding; Ubiquitin-like modifier; Autophagy; Vesicle

Chromosomal Location of Human Ortholog: 20q11.22

Cellular Component: microtubule; extrinsic to membrane; late endosome; autophagic vacuole; cytoplasmic vesicle; cytosol

Molecular Function:protein binding; phosphatidylethanolamine binding; microtubule binding; GABA receptor binding; phospholipid binding

Biological Process: mitochondrion degradation; cellular response to nitrogen starvation; autophagic vacuole formation

NCBI Summary:MAP1A and MAP1B are microtubule-associated proteins which mediate the physical interactions between microtubules and components of the cytoskeleton. MAP1A and MAP1B each consist of a heavy chain subunit and multiple light chain subunits. The protein encoded by this gene is one of the light chain subunits and can associate with either MAP1A or MAP1B. Two transcript variants encoding different isoforms have been found for this gene. The expression of variant 1 is suppressed in many tumor cell lines, suggesting that may be involved in carcinogenesis. [provided by RefSeq, Feb 2012]
UniProt Code:Q9H492
NCBI GenInfo Identifier:85701362
NCBI Gene ID:84557
NCBI Accession:Q9H492.2
UniProt Secondary Accession:Q9H492,Q9GZQ8,
UniProt Related Accession:Q9H492
Molecular Weight:
NCBI Full Name:Microtubule-associated proteins 1A/1B light chain 3A
NCBI Synonym Full Names:microtubule associated protein 1 light chain 3 alpha
NCBI Official Symbol:MAP1LC3A  
NCBI Official Synonym Symbols:LC3; LC3A; ATG8E; MAP1ALC3; MAP1BLC3  
NCBI Protein Information:microtubule-associated proteins 1A/1B light chain 3A
UniProt Protein Name:Microtubule-associated proteins 1A/1B light chain 3A
UniProt Synonym Protein Names:Autophagy-related protein LC3 A; Autophagy-related ubiquitin-like modifier LC3 A; MAP1 light chain 3-like protein 1; MAP1A/MAP1B light chain 3 A; MAP1A/MAP1B LC3 A; Microtubule-associated protein 1 light chain 3 alpha
UniProt Gene Name:MAP1LC3A  
UniProt Entry Name:MLP3A_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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