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Human SIRP alpha ELISA Kit

SKU:
HUFI02856
Product Type:
ELISA Kit
Size:
96 Assays
Uniprot:
P78324
Sensitivity:
0.094ng/ml
Range:
0.156-10ng/ml
ELISA Type:
Sandwich
Synonyms:
SIRPa, BIT, CD172a, MFR, MYD-1, SHPS1, SIRPA, Bit, BITbrain-immunoglobulin-like molecule with tyrosine-based activation motifs, Brain Ig-like molecule with tyrosine-based activation motifs, CD172 antigen-like family member A, CD172a, CD172a antigen,
Reactivity:
Human
Research Area:
Cell Biology
€599
Frequently bought together:

Description

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Human SIRP alpha ELISA Kit

The Human SIRP Alpha ELISA Kit from Assay Genie is a cutting-edge tool designed for the precise measurement of SIRP Alpha levels in human serum, plasma, and cell culture supernatants. With its advanced technology, this kit ensures high sensitivity and specificity, guaranteeing accurate and reproducible results for a variety of research applications.SIRP Alpha is a key protein involved in regulating immune responses and cell signaling. Its dysregulation has been linked to various diseases, including cancer, autoimmune disorders, and inflammatory conditions.

By accurately measuring SIRP Alpha levels, researchers can gain valuable insights into disease pathogenesis and potential therapeutic interventions.With the Human SIRP Alpha ELISA Kit, researchers can confidently study the role of SIRP Alpha in disease development and progression, opening up new possibilities for targeted therapies and personalized medicine. Trust Assay Genie for high-quality ELISA kits that drive innovative research forward.

Product Name:Human SIRP alpha ELISA Kit
Product Code:HUFI02856
Size:96 Assays
Alias:SIRPa, BIT, CD172a, MFR, MYD-1, SHPS1, SIRPA, Bit, BITbrain-immunoglobulin-like molecule with tyrosine-based activation motifs
Detection method:Sandwich ELISA, Double Antibody
Application:This immunoassay kit allows for the in vitro quantitative determination of Human SIRPa 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 SIRPa and the recovery rates were calculated by comparing the measured value to the expected amount of Human SIRPa in samples.
MatrixRecovery range(%)Average(%)
serum(n=5)86-9692
EDTA plasma(n=5)86-10494
UFH plasma(n=5)86-10597
Linearity:The linearity of the kit was assayed by testing samples spiked with appropriate concentration of Human SIRPa and their serial dilutions. The results were demonstrated by the percentage of calculated concentration to the expected.
Sample1:21:41:8
serum(n=5)86-95%87-100%87-101%
EDTA plasma(n=5)84-95%83-100%82-91%
UFH plasma(n=5)86-97%85-92%80-95%
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
UniprotP78324
UniProt Protein Function:SHPS1: Immunoglobulin-like cell surface receptor for CD47. Acts as docking protein and induces translocation of PTPN6, PTPN11 and other binding partners from the cytosol to the plasma membrane. Supports adhesion of cerebellar neurons, neurite outgrowth and glial cell attachment. May play a key role in intracellular signaling during synaptogenesis and in synaptic function. Involved in the negative regulation of receptor tyrosine kinase-coupled cellular responses induced by cell adhesion, growth factors or insulin. Mediates negative regulation of phagocytosis, mast cell activation and dendritic cell activation. CD47 binding prevents maturation of immature dendritic cells and inhibits cytokine production by mature dendritic cells. Binds PTPN11 when tyrosine-phosphorylated, except in macrophages, where it primarily binds PTPN6. Binds GRB2 in vitro. Binds FGR. Binds JAK2 irrespective of its phosphorylation status and forms a stable complex. Binds SCAP1 and/or SCAP2. The resulting complex recruits FYB. Binds PTK2B. Ubiquitous. Highly expressed in brain. Detected on myeloid cells, but not T-cells. Detected at lower levels in heart, placenta, lung, testis, ovary, colon, liver, small intestine, prostate, spleen, kidney, skeletal muscle and pancreas. 3 isoforms of the human protein are produced by alternative splicing.
UniProt Protein Details:Protein type:Cell surface; Motility/polarity/chemotaxis; Cell adhesion; Receptor, misc.; Membrane protein, integral

Chromosomal Location of Human Ortholog: 20p13

Cellular Component: membrane; integral to membrane; plasma membrane

Molecular Function:SH3 domain binding

Biological Process: blood coagulation; cell adhesion; leukocyte migration

NCBI Summary:The protein encoded by this gene is a member of the signal-regulatory-protein (SIRP) family, and also belongs to the immunoglobulin superfamily. SIRP family members are receptor-type transmembrane glycoproteins known to be involved in the negative regulation of receptor tyrosine kinase-coupled signaling processes. This protein can be phosphorylated by tyrosine kinases. The phospho-tyrosine residues of this PTP have been shown to recruit SH2 domain containing tyrosine phosphatases (PTP), and serve as substrates of PTPs. This protein was found to participate in signal transduction mediated by various growth factor receptors. CD47 has been demonstrated to be a ligand for this receptor protein. This gene and its product share very high similarity with several other members of the SIRP family. These related genes are located in close proximity to each other on chromosome 20p13. Multiple alternatively spliced transcript variants have been determined for this gene. [provided by RefSeq, Jul 2008]
UniProt Code:P78324
NCBI GenInfo Identifier:91105764
NCBI Gene ID:140885
NCBI Accession:NP_001035111.1
UniProt Secondary Accession:P78324,O00683, O43799, Q8N517, Q8TAL8, Q9H0Z2, Q9UDX2 Q9UIJ6, Q9Y4U9, A2A2E1, A8K411, B2R6C3,
UniProt Related Accession:P78324
Molecular Weight:54,967 Da
NCBI Full Name:tyrosine-protein phosphatase non-receptor type substrate 1
NCBI Synonym Full Names:signal-regulatory protein alpha
NCBI Official Symbol:SIRPA
NCBI Official Synonym Symbols:BIT; MFR; P84; SIRP; MYD-1; SHPS1; CD172A; PTPNS1
NCBI Protein Information:tyrosine-protein phosphatase non-receptor type substrate 1; myd-1 antigen; inhibitory receptor SHPS-1; macrophage fusion receptor; CD172 antigen-like family member A; tyrosine phosphatase SHP substrate 1; brain-immunoglobulin-like molecule with tyrosine-b
UniProt Protein Name:Tyrosine-protein phosphatase non-receptor type substrate 1
UniProt Synonym Protein Names:Brain Ig-like molecule with tyrosine-based activation motifs; Bit; CD172 antigen-like family member A; Inhibitory receptor SHPS-1; Macrophage fusion receptor; MyD-1 antigen; Signal-regulatory protein alpha-1; Sirp-alpha-1; Signal-regulatory protein alpha-2; Sirp-alpha-2; Signal-regulatory protein alpha-3; Sirp-alpha-3; p84
Protein Family:Nonribosomal peptide synthetase
UniProt Gene Name:SIRPA
UniProt Entry Name:SHPS1_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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