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Human MAX / MYC Associated Factor X ELISA Kit (HUFI00662)

SKU:
HUFI00662
Product Type:
ELISA Kit
Size:
96 Assays
Uniprot:
P61244
Sensitivity:
0.094ng/ml
Range:
0.156-10ng/ml
ELISA Type:
Sandwich
Synonyms:
MAX, BHLHD4, Protein max, bHLHd5, bHLHd6, bHLHd7, bHLHd8, Class D basic helix-loop-helix protein 4, helix-loop-helix zipper protein, MAX protein, MYC associated factor X, Myc-associated factor X, orf1
Reactivity:
Human
Research Area:
Epigenetics and Nuclear Signaling
€599
Frequently bought together:

Description

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Human MAX/MYC Associated Factor X ELISA Kit

The Human MAX (Myc-associated factor X) ELISA Kit is a powerful tool for detecting and quantifying MAX levels in human samples such as serum, plasma, and cell culture supernatants. This kit offers exceptional sensitivity and specificity, ensuring accurate and consistent results for a variety of research purposes.MAX is a key regulator of cell growth and proliferation, playing a vital role in various cellular processes such as apoptosis and differentiation.

Dysregulation of MAX has been linked to numerous diseases including cancer, making it a valuable biomarker for studying disease mechanisms and developing targeted therapies.With its reliable performance and broad application range, the Human MAX (Myc-associated factor X) ELISA Kit is a valuable resource for researchers interested in furthering their understanding of MAX function and its implications in health and disease.

Product Name:Human MAX / MYC Associated Factor X ELISA Kit
Product Code:HUFI00662
Size:96 Assays
Alias:MAX, BHLHD4, Protein max, bHLHd5, bHLHd6, bHLHd7, bHLHd8, Class D basic helix-loop-helix protein 4, helix-loop-helix zipper protein, MAX protein, MYC associated factor X, Myc-associated factor X, orf1
Detection method:Sandwich ELISA, Double Antibody
Application:This immunoassay kit allows for the in vitro quantitative determination of Human MAX concentrations in serum plasma and other biological fluids.
Sensitivity:0.094ng/ml
Range:0.156-10ng/ml
Storage:4°C for 6 months
Note:For Research Use Only
Recovery:Matrices listed below were spiked with certain level of Human MAX and the recovery rates were calculated by comparing the measured value to the expected amount of Human MAX in samples.
MatrixRecovery range(%)Average(%)
serum(n=5)98-102100
EDTA plasma(n=5)90-10596
UFH plasma(n=5)87-10495
Linearity:The linearity of the kit was assayed by testing samples spiked with appropriate concentration of Human MAX and their serial dilutions. The results were demonstrated by the percentage of calculated concentration to the expected.
Sample1:21:41:8
serum(n=5)88-103%94-105%86-103%
EDTA plasma(n=5)85-97%83-101%84-101%
UFH plasma(n=5)85-94%84-93%82-99%
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
UniprotP61244
UniProt Protein Function:MAX: a transcription factor. Forms a DNA- binding protein complex with MYC or MAD. The MYC-MAX complex is a transcriptional activator, whereas the MAD-MAX complex is a repressor. CPG methylation of the recognition site greatly inhibits DNA binding, suggesting that DNA methylation may regulate the MYC/MAX complex in vivo. May repress transcription via the recruitment of a chromatin remodeling complex containing H3-K9 histone methyltransferase activity. Three alternatively spliced isoforms have been reported.
UniProt Protein Details:Protein type:DNA-binding; Transcription factor

Chromosomal Location of Human Ortholog: 14q23

Cellular Component: nucleoplasm; PML body; dendrite; cytoplasm; nucleus

Molecular Function:protein binding; protein homodimerization activity; sequence-specific DNA binding; protein heterodimerization activity; transcription coactivator activity; transcription cofactor activity; protein complex binding; transcription factor activity

Biological Process: transcription from RNA polymerase II promoter; neuron apoptosis; regulation of transcription, DNA-dependent; retina development in camera-type eye; protein complex assembly; response to axon injury; cellular response to starvation; response to insulin stimulus

NCBI Summary:The protein encoded by this gene is a member of the basic helix-loop-helix leucine zipper (bHLHZ) family of transcription factors. It is able to form homodimers and heterodimers with other family members, which include Mad, Mxi1 and Myc. Myc is an oncoprotein implicated in cell proliferation, differentiation and apoptosis. The homodimers and heterodimers compete for a common DNA target site (the E box) and rearrangement among these dimer forms provides a complex system of transcriptional regulation. Mutations of this gene have been reported to be associated with hereditary pheochromocytoma. A pseudogene of this gene is located on the long arm of chromosome 7. Alternative splicing results in multiple transcript variants. [provided by RefSeq, Aug 2012]
UniProt Code:P61244
NCBI GenInfo Identifier:47117704
NCBI Gene ID:4149
NCBI Accession:P61244.1
UniProt Secondary Accession:P61244,P25912, P52163, Q14803, Q96CY8, A6NH73, A8K265 A8K4G4, A8K824,
UniProt Related Accession:P61244
Molecular Weight:160
NCBI Full Name:Protein max
NCBI Synonym Full Names:MYC associated factor X
NCBI Official Symbol:MAX
NCBI Official Synonym Symbols:bHLHd4
NCBI Protein Information:protein max; class D basic helix-loop-helix protein 4
UniProt Protein Name:Protein max
UniProt Synonym Protein Names:Class D basic helix-loop-helix protein 4; bHLHd4; Myc-associated factor X
Protein Family:Protein max
UniProt Gene Name:MAX
UniProt Entry Name:MAX_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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