Human Immunology ELISA Kits 9

Human TGF-beta-activated kinase 1 and MAP3K7-binding protein 1 (TAB1) ELISA Kit

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SKU:
HUEB2383
Product Type:
ELISA Kit
Size:
96 Assays
Uniprot:
Q15750
Range:
0.156-10 ng/mL
ELISA Type:
Sandwich
Synonyms:
TAB1, mitogen-activated protein kinase kinase kinase 7 interacting protein 1Mitogen-activated protein kinase kinase kinase 7-interacting protein 1, TAK1-binding protein 1
Reactivity:
Human

Description

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Product Name:Human TGF-beta-activated kinase 1 and MAP3K7-binding protein 1 (TAB1) ELISA Kit
Product Code:HUEB2383
Alias:TGF-beta-activated kinase 1 and MAP3K7-binding protein 1, Mitogen-activated protein kinase kinase kinase 7-interacting protein 1, TGF-beta-activated kinase 1-binding protein 1, TAK1-binding protein 1, TAB1, MAP3K7IP1
Uniprot:Q15750
Reactivity:Human
Range:0.156-10 ng/mL
Detection Method:Sandwich
Size:96 Assay
Storage:Please see kit components below for exact storage details
Note:For research use only
UniProt Protein Function:TAB1: May be an important signaling intermediate between TGFB receptors and MAP3K7/TAK1. May play an important role in mammalian embryogenesis. Interacts with XIAP and BIRC7. Interacts with TRAF6 and MAP3K7; during IL-1 signaling. Identified in the TRIKA2 complex composed of MAP3K7, TAB1 and TAB2. Ubiquitous. 2 isoforms of the human protein are produced by alternative splicing.
UniProt Protein Details:

Protein type:Activator; Motility/polarity/chemotaxis

Chromosomal Location of Human Ortholog: 22q13.1

Cellular Component: nucleoplasm; protein complex; cytoplasm; endosome membrane; cytosol

Molecular Function:protein binding; mitogen-activated protein kinase p38 binding; enzyme activator activity; protein complex binding; kinase activator activity; catalytic activity

Biological Process: heart morphogenesis; I-kappaB kinase/NF-kappaB cascade; activation of MAPK activity; in utero embryonic development; MyD88-independent toll-like receptor signaling pathway; stress-activated MAPK cascade; toll-like receptor 3 signaling pathway; toll-like receptor 10 signaling pathway; toll-like receptor 2 signaling pathway; activation of NF-kappaB transcription factor; toll-like receptor 5 signaling pathway; MyD88-dependent toll-like receptor signaling pathway; transforming growth factor beta receptor signaling pathway; toll-like receptor signaling pathway; activation of MAPKKK activity; JNK cascade; innate immune response; toll-like receptor 9 signaling pathway; toll-like receptor 4 signaling pathway; lung development

NCBI Summary:The protein encoded by this gene was identified as a regulator of the MAP kinase kinase kinase MAP3K7/TAK1, which is known to mediate various intracellular signaling pathways, such as those induced by TGF beta, interleukin 1, and WNT-1. This protein interacts and thus activates TAK1 kinase. It has been shown that the C-terminal portion of this protein is sufficient for binding and activation of TAK1, while a portion of the N-terminus acts as a dominant-negative inhibitor of TGF beta, suggesting that this protein may function as a mediator between TGF beta receptors and TAK1. This protein can also interact with and activate the mitogen-activated protein kinase 14 (MAPK14/p38alpha), and thus represents an alternative activation pathway, in addition to the MAPKK pathways, which contributes to the biological responses of MAPK14 to various stimuli. Alternatively spliced transcript variants encoding distinct isoforms have been reported. [provided by RefSeq, Jul 2008]
UniProt Code:Q15750
NCBI GenInfo Identifier:10720303
NCBI Gene ID:10454
NCBI Accession:Q15750.1
UniProt Secondary Accession:Q15750,Q2PP09, Q8IZW2,
UniProt Related Accession:Q15750
Molecular Weight:504
NCBI Full Name:TGF-beta-activated kinase 1 and MAP3K7-binding protein 1
NCBI Synonym Full Names:TGF-beta activated kinase 1/MAP3K7 binding protein 1
NCBI Official Symbol:TAB1  
NCBI Official Synonym Symbols:3'-Tab1; MAP3K7IP1  
NCBI Protein Information:TGF-beta-activated kinase 1 and MAP3K7-binding protein 1; TAK1-binding protein 1; transforming growth factor beta-activated kinase-binding protein 1; mitogen-activated protein kinase kinase kinase 7-interacting protein 1
UniProt Protein Name:TGF-beta-activated kinase 1 and MAP3K7-binding protein 1
UniProt Synonym Protein Names:Mitogen-activated protein kinase kinase kinase 7-interacting protein 1; TGF-beta-activated kinase 1-binding protein 1; TAK1-binding protein 1
Protein Family:TGF-beta-activated kinase 1 and MAP3K7-binding protein
UniProt Gene Name:TAB1  
UniProt Entry Name:TAB1_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 TAB1 / TGF-beta Activated Kinase 1 / MAP3K7 Binding Protein 1 ELISA Kit
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