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Human Fermitin family homolog 2 (FERMT2) ELISA Kit (HUEB2369)

SKU:
HUEB2369
Product Type:
ELISA Kit
Size:
96 Assays
Uniprot:
Q96AC1
ELISA Type:
Sandwich
Reactivity:
Human
$779
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Description

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Human Fermitin family homolog 2 (FERMT2) ELISA Kit

The Human Fermitin Family Homolog 2 (FERMT2) ELISA Kit is a cutting-edge tool for the precise quantification of FERMT2 levels in human samples including serum, plasma, and cell culture supernatants. This innovative kit offers exceptional sensitivity and specificity, ensuring robust and consistent results for a wide range of research applications.FERMT2, also known as Kindlin-2, is a critical protein involved in cell adhesion and signaling pathways. It plays a key role in various cellular processes such as cell migration, proliferation, and differentiation.

Dysregulation of FERMT2 has been linked to several diseases including cancer, cardiovascular disorders, and inflammatory conditions, making it a valuable biomarker for studying these diseases and potential therapeutic interventions.Overall, the Human Fermitin Family Homolog 2 (FERMT2) ELISA Kit is a powerful tool for researchers seeking to explore the role of FERMT2 in health and disease, providing accurate and reliable measurements to advance scientific understanding and clinical applications.

Product Name:Human Fermitin family homolog 2 (FERMT2) ELISA Kit
SKU:HUEB2369
Size:96T
Target:Human Fermitin family homolog 2 (FERMT2)
Synonyms:Kindlin-2, Mitogen-inducible gene 2 protein, Pleckstrin homology domain-containing family C member 1, MIG-2, PH domain-containing family C member 1, KIND2, MIG2, PLEKHC1
Assay Type:Sandwich
Detection Method:ELISA
Reactivity:Human
Detection Range:78-5000pg/mL
Sensitivity:39.2pg/mL
Intra CV:6.0%
Inter CV:7.3%
Linearity:
Sample1:21:41:81:16
Serum(N=5)101-111%107-117%111-120%105-115%
EDTA Plasma(N=5)101-111%108-118%109-118%82-95%
Heparin Plasma(N=5)91-100%89-102%99-109%102-113%
Recovery:
Sample TypeAverage(%)Recovery Range(%)
Serum9791-103
Plasma9993-105
Function:Scaffolding protein that enhances integrin activation mediated by TLN1 and/or TLN2, but activates integrins only weakly by itself. Binds to membranes enriched in phosphoinositides. Enhances integrin-mediated cell adhesion onto the extracellular matrix and cell spreading; this requires both its ability to interact with integrins and with phospholipid membranes. Required for the assembly of focal adhesions. Participates in the connection between extracellular matrix adhesion sites and the actin cytoskeleton and also in the orchestration of actin assembly and cell shape modulation. Recruits FBLIM1 to focal adhesions. Plays a role in the TGFB1 and integrin signaling pathways. Stabilizes active CTNNB1 and plays a role in the regulation of transcription mediated by CTNNB1 and TCF7L2/TCF4 and in Wnt signaling.
Uniprot:Q96AC1
Sample Type:Serum, plasma, tissue homogenates, cell culture supernates and other biological fluids
Specificity:Natural and recombinant human Fermitin family homolog 2
Sub Unit:Interacts with ILK (By similarity). Interacts with FBLIM1. Interacts with ITGB1 and ITGB3. Interacts with active, unphosphorylated CTNNB1. Identified in a complex with CTNNB1 and TCF7L2/TCF4. Interacts with ITGB1; the interaction is inhibited in presence of ITGB1BP1.
Subcellular Location:Cytoplasm Cytoplasm Cell cortex Cytoplasm Cytoskeleton Cell junction Focal adhesion Membrane Peripheral membrane protein Cytoplasmic side Cell projection Lamellipodium membrane Peripheral membrane protein Cytoplasmic side Nucleus Cytoplasm Myofibril Sarcomere I band Cell surface Colocalizes with actin stress fibers at cell-ECM focal adhesion sites. Colocalizes with ITGB3 at lamellipodia at the leading edge of spreading cells. Binds to membranes that contain phosphatidylinositides.
Storage:Please see kit components below for exact storage details
Note:For research use only
UniProt Protein Function:kindlin-2: Participates in the connection between ECM adhesion sites and the actin cytoskeleton and also in the orchestration of actin assembly and cell shape modulation. Recruits migfilin (FBLP1) protein to cell-ECM focal adhesion sites. Belongs to the kindlin family. 3 isoforms of the human protein are produced by alternative splicing.
UniProt Protein Details:

Protein type:Motility/polarity/chemotaxis; Cytoskeletal

Chromosomal Location of Human Ortholog: 14q22.1

Cellular Component: cytoplasm; cytosol; extrinsic to internal side of plasma membrane; focal adhesion; nucleoplasm; nucleus

Molecular Function:phosphatidylinositol-3,4,5-triphosphate binding; protein binding

Biological Process: cell-matrix adhesion; focal adhesion formation; integrin activation; integrin-mediated signaling pathway; transforming growth factor beta receptor signaling pathway; Wnt receptor signaling pathway

UniProt Code:Q96AC1
NCBI GenInfo Identifier:38258220
NCBI Gene ID:10979
NCBI Accession:Q96AC1.1
UniProt Secondary Accession:Q96AC1,Q14840, Q86TY7, B5TJY2,
UniProt Related Accession:Q96AC1
Molecular Weight:78,675 Da
NCBI Full Name:Fermitin family homolog 2
NCBI Synonym Full Names:fermitin family member 2
NCBI Official Symbol:FERMT2
NCBI Official Synonym Symbols:MIG2; KIND2; mig-2; UNC112; PLEKHC1; UNC112B
NCBI Protein Information:fermitin family homolog 2
UniProt Protein Name:Fermitin family homolog 2
UniProt Synonym Protein Names:Kindlin-2; Mitogen-inducible gene 2 protein; MIG-2; Pleckstrin homology domain-containing family C member 1; PH domain-containing family C member 1
Protein Family:Fermitin family
UniProt Gene Name:FERMT2
UniProt Entry Name:FERM2_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.

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