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Mouse Pancreas/duodenum homeobox protein 1 (Pdx1) ELISA Kit (MOEB2350)

SKU:
MOEB2350
Product Type:
ELISA Kit
Size:
96 Assays
Uniprot:
P52946
ELISA Type:
Sandwich
Reactivity:
Mouse
€649
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Description

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Mouse Pancreas/duodenum homeobox protein 1 (Pdx1) ELISA Kit

The Mouse Pancreas/Duodenum Homeobox Protein 1 (PDX1) ELISA Kit is specifically designed for the accurate and sensitive detection of PDX1 levels in mouse samples, including serum, plasma, and cell culture supernatants. This kit offers high specificity and reliability, ensuring reproducible results for a variety of research applications.PDX1 is a critical transcription factor involved in the development and function of the pancreas and duodenum.

It plays a key role in regulating the expression of genes involved in insulin production, making it a valuable biomarker for studying diabetes and pancreatic disorders. The Mouse PDX1 ELISA Kit provides researchers with a powerful tool for studying the role of PDX1 in health and disease, aiding in the development of potential therapies for pancreatic-related conditions.

Product Name:Mouse Pancreas/duodenum homeobox protein 1 (Pdx1) ELISA Kit
SKU:MOEB2350
Size:96T
Target:Mouse Pancreas/duodenum homeobox protein 1 (Pdx1)
Synonyms:Insulin promoter factor 1, Islet/duodenum homeobox 1, Somatostatin-transactivating factor 1, IPF-1, IDX-1, STF-1, Ipf1
Assay Type:Sandwich
Detection Method:ELISA
Reactivity:Mouse
Intra CV:0.0%
Inter CV:0.0%
Linearity:
Sample1:21:41:81:16
Serum(N=5)107-117%101-111%108-117%103-112%
EDTA Plasma(N=5)100-111%80-92%117-117%105-114%
Heparin Plasma(N=5)106-115%86-98%85-94%82-91%
Recovery:
Sample TypeAverage(%)Recovery Range(%)
Serum8080-80
Plasma8080-80
Function:Activates insulin and somatostatin gene transcription. Key regulator of islet peptide hormone expression but also responsible for the development of the pancreas, most probably by determining maturation and differentiation of common pancreatic precursor cells in the developing gut. As part of a PDX1:PBX1b:MEIS2b complex in pancreatic acinar cells is involved in the transcriptional activation of the ELA1 enhancer; the complex binds to the enhancer B element and cooperates with the transcription factor 1 complex (PTF1) bound to the enhancer A element. Binds the DNA sequence 5'-CC[CT]TAATGGG-3'.
Uniprot:P52946
Sample Type:Serum, plasma, tissue homogenates, cell culture supernates and other biological fluids
Specificity:Natural and recombinant mouse Pancreas/duodenum homeobox protein 1
Sub Unit:Interacts with the basic helix-loop-helix domains of TCF3(E47) and NEUROD1 and with HMG-I(Y). Interacts with the methyltransferase SETD7 (By similarity). Interacts with SPOP. Part of a PDX1:PBX1b:MEIS2b complex.
Subcellular Location:Nucleus Cytoplasm Cytosol
Storage:Please see kit components below for exact storage details
Note:For research use only
UniProt Protein Function:PDX1: Activates insulin, somatostatin, glucokinase, islet amyloid polypeptide and glucose transporter type 2 gene transcription. Particularly involved in glucose-dependent regulation of insulin gene transcription. Binds preferentially the DNA motif 5'-[CT]TAAT[TG]-3'. During development, specifies the early pancreatic epithelium, permitting its proliferation, branching and subsequent differentiation. At adult stage, required for maintaining the hormone-producing phenotype of the beta-cell. Interacts with the basic helix-loop-helix domains of TCF3(E47) and NEUROD1 and with HMG-I(Y). Interacts with SPOP. Interacts with the methyltransferase SETD7. Duodenum and pancreas (Langerhans islet beta cells and small subsets of endocrine non-beta-cells, at low levels in acinar cells). Belongs to the Antp homeobox family. IPF1/XlHbox-8 subfamily.Protein type: Transcription factor; DNA-bindingCellular Component: cytoplasm; intracellular; nuclear speck; nucleusMolecular Function: chromatin binding; DNA binding; protein binding; protein complex binding; protein heterodimerization activity; RNA polymerase II transcription factor activity, enhancer binding; sequence-specific DNA binding; transcription factor activity; transcription factor bindingBiological Process: cell differentiation; detection of glucose; endocrine pancreas development; exocrine pancreas development; glucose homeostasis; glucose metabolic process; gut development; insulin secretion; liver development; morphogenesis of embryonic epithelium; negative regulation of cell proliferation; negative regulation of transcription from RNA polymerase II promoter; negative regulation of transcription, DNA-dependent; nitric oxide mediated signal transduction; pancreas development; positive regulation of cell proliferation; positive regulation of DNA binding; positive regulation of insulin secretion; positive regulation of transcription from RNA polymerase II promoter; positive regulation of transcription, DNA-dependent; regulation of cell proliferation; regulation of gene expression; regulation of transcription from RNA polymerase II promoter; response to glucose stimulus; smoothened signaling pathway; stem cell differentiation; transcription from RNA polymerase II promoter; transcription, DNA-dependent
UniProt Protein Details:
NCBI Summary:
UniProt Code:P52946
NCBI GenInfo Identifier:1708541
NCBI Gene ID:18609
NCBI Accession:P52946.1
UniProt Secondary Accession:P52946,Q3ZB03
UniProt Related Accession:P52946
Molecular Weight:30,999 Da
NCBI Full Name:Pancreas/duodenum homeobox protein 1
NCBI Synonym Full Names:pancreatic and duodenal homeobox 1
NCBI Official Symbol:Pdx1
NCBI Official Synonym Symbols:Ipf1; IDX-1; IPF-1; Mody4; STF-1; pdx-1
NCBI Protein Information:pancreas/duodenum homeobox protein 1
UniProt Protein Name:Pancreas/duodenum homeobox protein 1
UniProt Synonym Protein Names:Insulin promoter factor 1; IPF-1; Islet/duodenum homeobox 1; IDX-1; Somatostatin-transactivating factor 1; STF-1
Protein Family:Pyridoxal 5'-phosphate synthase
UniProt Gene Name:Pdx1
UniProt Entry Name:PDX1_MOUSE
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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