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Human CD44 ELISA Kit

SKU:
HUFI00472
Product Type:
ELISA Kit
Size:
96 Assays
Uniprot:
P16070
Sensitivity:
0.094ng/ml
Range:
0.156-10ng/ml
ELISA Type:
Sandwich
Synonyms:
CD44, ECMR-III, HCAM, HCELL, LHR, MDU2, MDU3, MIC4, MUTCH-I, Pgp1, CD44R, CDw44, CSPG8, Epican, HUTCH-I, Hyaluronate receptor, IN, LHR, MC56, PGP-1, PGP-I, Phagocytic glycoprotein 1, Phagocytic glycoprotein I, IN
Reactivity:
Human
Research Area:
Cell Biology
$719
Frequently bought together:

Description

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Human CD44 ELISA Kit

The Human CD44 ELISA Kit is a powerful tool for the precise measurement of CD44 levels in human samples such as serum, plasma, and cell culture supernatants. This kit offers high sensitivity and specificity, guaranteeing accurate and consistent results for a variety of research purposes.CD44 is a cell surface glycoprotein involved in cell-cell interactions, cell adhesion, and migration. It plays a crucial role in various physiological and pathological processes, including inflammation, immune response, and cancer metastasis.

The ability to accurately measure CD44 levels can provide valuable insights into these processes and facilitate the development of novel therapeutic strategies.The Human CD44 ELISA Kit is user-friendly and versatile, suitable for use in laboratories conducting research on cancer, immunology, and cell biology. By accurately quantifying CD44 levels, researchers can better understand its role in disease pathogenesis and potentially identify new targets for intervention.

Product Name:Human CD44 ELISA Kit
Product Code:HUFI00472
Size:96 Assays
Alias:CD44, ECMR-III, HCAM, HCELL, LHR, MDU2, MDU3, MIC4, MUTCH-I, Pgp1, CD44R, CDw44, CSPG8, Epican, HUTCH-I, Hyaluronate receptor, IN, LHR, MC56, PGP-1, PGP-I, Phagocytic glycoprotein 1, Phagocytic glycoprotein I, IN
Detection method:Sandwich ELISA, Double Antibody
Application:This immunoassay kit allows for the in vitro quantitative determination of Human CD44 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 CD44 and the recovery rates were calculated by comparing the measured value to the expected amount of Human CD44 in samples.
MatrixRecovery range(%)Average(%)
serum(n=5)88-10597
EDTA plasma(n=5)88-10294
UFH plasma(n=5)90-10495
Linearity:The linearity of the kit was assayed by testing samples spiked with appropriate concentration of Human CD44 and their serial dilutions. The results were demonstrated by the percentage of calculated concentration to the expected.
Sample1:21:41:8
serum(n=5)85-105%87-103%85-96%
EDTA plasma(n=5)85-98%88-99%82-97%
UFH plasma(n=5)86-96%80-100%80-99%
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
UniprotP16070
UniProt Protein Function:CD44: Receptor for hyaluronic acid (HA). Mediates cell-cell and cell-matrix interactions through its affinity for HA, and possibly also through its affinity for other ligands such as osteopontin, collagens, and matrix metalloproteinases (MMPs). Adhesion with HA plays an important role in cell migration, tumor growth and progression. Also involved in lymphocyte activation, recirculation and homing, and in hematopoiesis. Altered expression or dysfunction causes numerous pathogenic phenotypes. Great protein heterogeneity due to numerous alternative splicing and post-translational modification events. Interacts with PKN2. Interacts with HA, as well as other glycosaminoglycans, collagen, laminin, and fibronectin via its N-terminal segment. Interacts with ANK, the ERM proteins (VIL2, RDX and MSN), and NF2 via its C-terminal segment. Isoform 10 (epithelial isoform) is expressed by cells of epithelium and highly expressed by carcinomas. Expression is repressed in neuroblastoma cells. 19 isoforms of the human protein are produced by alternative splicing.
UniProt Protein Details:Protein type:Cell adhesion; Motility/polarity/chemotaxis; Membrane protein, integral; Receptor, misc.

Chromosomal Location of Human Ortholog: 11p13

Cellular Component: Golgi apparatus; focal adhesion; cell surface; basolateral plasma membrane; integral to plasma membrane; cytoplasm; plasma membrane; external side of plasma membrane

Molecular Function:collagen binding; hematopoietin/interferon-class (D200-domain) cytokine receptor activity; protein binding; hyaluronic acid binding; hyalurononglucosaminidase activity

Biological Process: extracellular matrix organization and biogenesis; Wnt receptor signaling pathway; positive regulation of heterotypic cell-cell adhesion; glycosaminoglycan metabolic process; cytokine and chemokine mediated signaling pathway; cell-matrix adhesion; pathogenesis; negative regulation of caspase activity; positive regulation of peptidyl-serine phosphorylation; hyaluronan catabolic process; extracellular matrix disassembly; negative regulation of DNA damage response, signal transduction by p53 class mediator; cell-cell adhesion; positive regulation of peptidyl-tyrosine phosphorylation; ureteric bud branching; cartilage development; carbohydrate metabolic process; blood coagulation; leukocyte migration; healing during inflammatory response; hyaluronan metabolic process; negative regulation of apoptosis

Disease: Blood Group, Indian System

NCBI Summary:The protein encoded by this gene is a cell-surface glycoprotein involved in cell-cell interactions, cell adhesion and migration. It is a receptor for hyaluronic acid (HA) and can also interact with other ligands, such as osteopontin, collagens, and matrix metalloproteinases (MMPs). This protein participates in a wide variety of cellular functions including lymphocyte activation, recirculation and homing, hematopoiesis, and tumor metastasis. Transcripts for this gene undergo complex alternative splicing that results in many functionally distinct isoforms, however, the full length nature of some of these variants has not been determined. Alternative splicing is the basis for the structural and functional diversity of this protein, and may be related to tumor metastasis. [provided by RefSeq, Jul 2008]
UniProt Code:P16070
NCBI GenInfo Identifier:308153615
NCBI Gene ID:960
NCBI Accession:P16070.3
UniProt Secondary Accession:P16070,O95370, P22511, Q04858, Q13419, Q13957, Q13958 Q13959, A5YRN9, B6EAT9, D3DR12, D3DR13,
UniProt Related Accession:P16070
Molecular Weight:81,538 Da
NCBI Full Name:CD44 antigen
NCBI Synonym Full Names:CD44 molecule (Indian blood group)
NCBI Official Symbol:CD44 Â 
NCBI Official Synonym Symbols:IN; LHR; MC56; MDU2; MDU3; MIC4; Pgp1; CDW44; CSPG8; HCELL; HUTCH-I; ECMR-III Â 
NCBI Protein Information:CD44 antigen; epican; Hermes antigen; hyaluronate receptor; phagocytic glycoprotein 1; heparan sulfate proteoglycan; cell surface glycoprotein CD44; extracellular matrix receptor III; chondroitin sulfate proteoglycan 8; GP90 lymphocyte homing/adhesion receptor; hematopoietic cell E- and L-selectin ligand; homing function and Indian blood group system
UniProt Protein Name:CD44 antigen
UniProt Synonym Protein Names:CDw44; Epican; Extracellular matrix receptor III; ECMR-III; GP90 lymphocyte homing/adhesion receptor; HUTCH-I; Heparan sulfate proteoglycan; Hermes antigen; Hyaluronate receptor; Phagocytic glycoprotein 1; PGP-1; Phagocytic glycoprotein I
Protein Family:CD44 antigen
UniProt Gene Name:CD44 Â 
UniProt Entry Name:CD44_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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