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Human Microtubule-associated proteins 1A/1B light chain 3B (MAP1LC3B) ELISA Kit (HUEB2380)

SKU:
HUEB2380
Product Type:
ELISA Kit
Size:
96 Assays
Uniprot:
Q9GZQ8
Range:
0.156-10 ng/mL
ELISA Type:
Sandwich
Synonyms:
MAP1LC3B, Microtubule-associated protein 1 light chain 3 beta, MAP1A
Reactivity:
Human
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Description

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Human Microtubule-associated proteins 1A/1B light chain 3B (MAP1LC3B) ELISA Kit

The Human Microtubule-Associated Proteins 1A/1B Light Chain 3B (MAP1LC3B) ELISA Kit is specifically designed for the precise measurement of MAP1LC3B levels in human samples including serum, plasma, and cell culture supernatants. This advanced kit offers exceptional sensitivity and accuracy, guaranteeing dependable and consistent results for a variety of research purposes.MAP1LC3B is a key protein involved in the regulation of autophagy, a crucial cellular process responsible for maintaining cellular homeostasis and eliminating damaged organelles and proteins.

Dysregulation of autophagy has been linked to various diseases including cancer, neurodegenerative disorders, and inflammatory conditions, highlighting the importance of MAP1LC3B as a biomarker for understanding these diseases and potentially developing novel therapies.Overall, the Human MAP1LC3B ELISA Kit provides researchers with a valuable tool for studying autophagy-related pathways and investigating the role of MAP1LC3B in disease pathogenesis, ultimately contributing to advancements in biomedical research and therapeutic development.

Product Name:Human Microtubule-associated proteins 1A/1B light chain 3B (MAP1LC3B) ELISA Kit
SKU:HUEB2380
Size:96T
Target:Human Microtubule-associated proteins 1A/1B light chain 3B (MAP1LC3B)
Synonyms:Autophagy-related protein LC3 B, Autophagy-related ubiquitin-like modifier LC3 B, MAP1 light chain 3-like protein 2, MAP1A/MAP1B light chain 3 B, Microtubule-associated protein 1 light chain 3 beta, MAP1A/MAP1B LC3 B, MAP1ALC3
Assay Type:Sandwich
Detection Method:ELISA
Reactivity:Human
Detection Range:0.156-10ng/mL
Sensitivity:0.078ng/mL
Intra CV:5.3%
Inter CV:7.5%
Linearity:
Sample1:21:41:81:16
Serum(N=5)100-112%94-106%81-90%102-110%
EDTA Plasma(N=5)90-100%80-90%102-112%85-95%
Heparin Plasma(N=5)109-119%95-104%103-116%108-117%
Recovery:
Sample TypeAverage(%)Recovery Range(%)
Serum8580-91
Plasma8781-93
Function:Ubiquitin-like modifier involved in formation of autophagosomal vacuoles (autophagosomes). Plays a role in mitophagy which contributes to regulate mitochondrial quantity and quality by eliminating the mitochondria to a basal level to fulfill cellular energy requirements and preventing excess ROS production. Whereas LC3s are involved in elongation of the phagophore membrane, the GABARAP/GATE-16 subfamily is essential for a later stage in autophagosome maturation. Promotes primary ciliogenesis by removing OFD1 from centriolar satellites via the autophagic pathway.
Uniprot:Q9GZQ8
Sample Type:Serum, plasma, tissue homogenates, cell culture supernates and other biological fluids
Specificity:Natural and recombinant human Microtubule-associated proteins 1A/1B light chain 3B
Sub Unit:3 different light chains, LC1, LC2 and LC3, can associate with MAP1A and MAP1B proteins (By similarity). Interacts at microtubules with CABP1 (via EF-hands 1 and 2) but not with calmodulin (PubMed:15095872). Interacts with FYCO1 (via C-terminus) (PubMed:20100911, PubMed:24089205). Interacts with TP53INP1 and TP53INP2 (PubMed:22470510). Interacts with TBC1D25 (PubMed:21383079). Directly interacts with SQSTM1; this interaction leads to MAP1LC3B recruitment to inclusion bodies containing polyubiquitinated protein aggregates and to inclusion body degradation by autophagy (PubMed:17580304, PubMed:24089205, PubMed:18524774). Interacts with ATG4B, MAPK15 and BNIP3 (PubMed:22948227, PubMed:23209295). Interacts with MAPB1, KEAP1, PCM1, OFD1, CEP131, and TECPR2 (PubMed:20562859, PubMed:24089205). Interacts with TBC1D5 (PubMed:22354992). Found in a complex with UBQLN1 and UBQLN2 (PubMed:20529957). Interacts with UBQLN4 (via STI1 1 and 2 domains). Interacts with UBQLN1 in the presence of UBQLN4 (PubMed:23459205). Interacts with ATG13 (PubMed:24290141). Interacts with FAM134A, FAM134B and FAM134C (PubMed:26040720).
Research Area:Cancer
Subcellular Location:Cytoplasm Cytoskeleton Endomembrane system Lipid-anchor Cytoplasmic vesicle Autophagosome membrane Lipid-anchor Cytoplasmic vesicle Autophagosome LC3-II binds to the autophagic membranes. Localizes also to discrete punctae along the ciliary axoneme (By similarity).
Storage:Please see kit components below for exact storage details
Note:For research use only
NCBI Summary:The product of this gene is a subunit of neuronal microtubule-associated MAP1A and MAP1B proteins, which are involved in microtubule assembly and important for neurogenesis. Studies on the rat homolog implicate a role for this gene in autophagy, a process that involves the bulk degradation of cytoplasmic component. [provided by RefSeq, Jul 2008]
UniProt Code:Q9GZQ8
NCBI GenInfo Identifier:17433141
NCBI Gene ID:81631
NCBI Accession:Q9GZQ8.3
UniProt Related Accession:Q9GZQ8
Molecular Weight:
NCBI Full Name:Microtubule-associated proteins 1A/1B light chain 3B
NCBI Synonym Full Names:microtubule associated protein 1 light chain 3 beta
NCBI Official Symbol:MAP1LC3B
NCBI Official Synonym Symbols:LC3B; ATG8F; MAP1LC3B-a; MAP1A/1BLC3
NCBI Protein Information:microtubule-associated proteins 1A/1B light chain 3B
UniProt Protein Name:Microtubule-associated proteins 1A/1B light chain 3B
UniProt Synonym Protein Names:Autophagy-related protein LC3 B; Autophagy-related ubiquitin-like modifier LC3 B; MAP1 light chain 3-like protein 2; MAP1A/MAP1B light chain 3 B; MAP1A/MAP1B LC3 B; Microtubule-associated protein 1 light chain 3 beta
UniProt Gene Name:MAP1LC3B
UniProt Entry Name:MLP3B_HUMAN
Component Quantity (96 Assays) Storage
ELISA Microplate (Dismountable) 8×12 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 120µL -20°C
Detection Reagent B 120µL -20°C
Wash Buffer 30mL 4°C
Substrate 10mL 4°C
Stop Solution 10mL 4°C
Plate Sealer 5 -

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

*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
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.

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.
ELISA
Human LC3B / MAP1LC3B ELISA Kit

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