Name: Human cluster of differentiation 63,CD63 ELISA Kit
Brand: CUSABIO
SKU: CSB-E14107h

Product Details

Target Name

CD63 molecule

Alternative Names

CD63; MLA1; TSPAN30; CD63 antigen; Granulophysin; Lysosomal-associated membrane protein 3; LAMP-3; Lysosome integral membrane protein 1; Limp1; Melanoma-associated antigen ME491; OMA81H; Ocular melanoma-associated antigen; Tetraspanin-30; Tspan-30; CD antigen CD63

Abbreviation

CD63

Uniprot No.

P08962

Species

Homo sapiens (Human)

Sample Types

serum, plasma, tissue homogenates

Detection Range

0.625 ng/mL-40 ng/mL

Sensitivity

0.156 ng/mL

Assay Time

1-5h

Sample Volume

50-100ul

Detection Wavelength

450 nm

Research Area

Immunology

Assay Principle

quantitative

Measurement

Sandwich

Precision

Linearity

To assess the linearity of the assay, samples were spiked with high concentrations of human CD63 in various matrices and diluted with the Sample Diluent to produce samples with values within the dynamic range of the assay.
	Sample	Serum(n=4)
1:1	Average %	96
	Range %	92-104
1:2	Average %	87
	Range %	81-94
1:4	Average %	98
	Range %	92-106
1:8	Average %	96
	Range %	92-103

Recovery

The recovery of human CD63 spiked to levels throughout the range of the assay in various matrices was evaluated. Samples were diluted prior to assay as directed in the Sample Preparation section.

Sample Type	Average % Recovery	Range
Serum (n=5)	104	98-107
EDTA plasma (n=4)	93	89-103

Typical Data

These standard curves are provided for demonstration only. A standard curve should be generated for each set of samples assayed.

ng/ml	OD1	OD2	Average	Corrected
40	2.639	2.538	2.589	2.428
20	1.947	1.846	1.897	1.736
10	1.301	1.312	1.307	1.146
5	0.776	0.765	0.771	0.610
2.5	0.473	0.462	0.468	0.307
1.25	0.336	0.347	0.342	0.181
0.625	0.246	0.240	0.243	0.082
0	0.162	0.160	0.161

Materials provided

A micro ELISA plate --- The 96-well plate has been pre-coated with an anti-human CD63 antibody. This dismountable microplate can be divided into 12 x 8 strip plates.
Two vials lyophilized standard ---Dilute a bottle of the standard at dilution series, read the OD values, and then draw a standard curve.
One vial Biotin-labeled CD63 antibody (100 x concentrate) (120 μl/bottle) ---Act as the detection antibody.
One vial HRP-avidin (100 x concentrate) (120 μl/bottle) ---Bind to the detection antibody and react with the TMB substrate to make the solution chromogenic.
One vial Biotin-antibody Diluent (15 ml/bottle) ---Dilute the Biotin-antibody.
One vial HRP-avidin Diluent (15 ml/bottle) ---Dilute the HRP-avidin solution.
One vial Sample Diluent (50 ml/bottle)---Dilute the sample to an appropriate concentration.
One vial Wash Buffer (25 x concentrate) (20 ml/bottle) ---Wash away unbound or free substances.
One vial TMB Substrate (10 ml/bottle) ---Act as the chromogenic agent. TMB interacts with HRP, eliciting the solution turns blue.
One vial Stop Solution (10 ml/bottle) ---Stop the color reaction. The solution color immediately turns from blue to yellow.
Four Adhesive Strips (For 96 wells) --- Cover the microplate when incubation.
An instruction manual

Materials not provided

A microplate reader capable of measuring absorbance at 450 nm, with the correction wavelength set at 540 nm or 570 nm.
An incubator can provide stable incubation conditions up to 37°C±5°C.
Centrifuge
Vortex
Squirt bottle, manifold dispenser, or automated microplate washer
Absorbent paper for blotting the microtiter plate
50-300ul multi-channel micropipette
Pipette tips
Single-channel micropipette with different ranges
100ml and 500ml graduated cylinders
Deionized or distilled water
Timer
Test tubes for dilution

ELISA Data Analysis

ELISA Standard Curve & Curve Expert software

Troubleshooting
and FAQs

ELISA kit FAQs

Storage

Store at 2-8°C. Please refer to protocol.

Shelf Life

6 months

Lead Time

3-5 working days after you place the order, and it takes another 3-5 days for delivery via DHL or FedEx.

Description

CD63 is a transmembrane glycoprotein belonging to the tetraspanin family, characterized by four transmembrane domains and two extracellular loops. This molecule is predominantly localized in late endosomes and lysosomes, where it plays important roles in vesicular trafficking, membrane fusion, and cellular signaling processes. CD63 serves as a widely recognized marker for exosomes and is involved in various physiological functions including immune cell activation, platelet aggregation, and tumor progression. Its expression and secretion patterns make it a valuable biomarker for studying cellular communication and disease states.

The Human cluster of differentiation 63,CD63 ELISA Kit (CSB-E14107h) uses a quantitative sandwich measurement approach to detect CD63 molecule in human samples. This assay accommodates serum, plasma, and tissue homogenates with a detection range of 0.625 ng/mL to 40 ng/mL and shows sensitivity of 0.156 ng/mL. The protocol requires 50-100ul sample volume and can be completed within 1-5 hours, with detection performed at 450 nm wavelength.

Application Examples

Note: The following application examples are drawn from a selection of publications citing this product. For additional applications, please refer to the full list of references in the "Citations" section.

This ELISA kit has been used in extracellular vesicle research to quantify CD63 levels in small extracellular vesicles and evaluate preservation effects on vesicle contents. The kit serves as a tool for characterizing tetraspanin markers in isolated vesicles and measuring their protein composition across different experimental conditions.

• Extracellular vesicle characterization - Quantification of CD63 tetraspanin levels in small extracellular vesicles to determine vesicle integrity and marker expression
• Preservation studies - Evaluation of CD63 protein levels in extracellular vesicles following various storage and preservation protocols
• Cell line-derived vesicle analysis - Detection and measurement of CD63 in extracellular vesicle preparations derived from different cellular sources
• Stem cell research - Quantification of CD63 in lysed extracellular vesicles from human amniotic fluid-derived stem cells for vesicle characterization studies

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Customer Reviews and Q&A

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■ Q&A

Please let me ask one question.
Can this kit detect only human CD63? Or is there some cross-reactivity with CD63 of other species(mouse, rat)?

Thanks for your inquiry.
#CSB-E14107h,Human cluster of differentiation 63,CD63 ELISA Kit
This kit is developed based on matrix of human and it is directed to human samples.Human CD63 is a different protein from rat/mouse CD63.The sequence of human CD63 and matrix are different from that of rat/mouse CD63.
And we did not do analysis of cross-reactivity on human DC63 and rat/mouse CD63.If you are going to test rat/mouse sample,the kit of respective species is advised.
If further questions,pls do ont hesitate to cotnact us.

I want to lyse my sample and I will use RIPA buffer for lysis.
Does RIPA buffer has any effect to ELISA result?

Thanks for your inquiry.
Usually RIPA would contains components SDS、Triton X-100、NP-40,ect.Pls kindly be noted that it must not contain SDS which would destroy the hydrogen bonds of protein macromolecules and causes denaturation of proteins .And Triton X-100、NP-40 should be controlled in the concentration range (usually not over 1% which was not validated ever).
For RIPA buffer,we did not ever use this kind of treatment method.You can only try by yourself and do pre-testing.On our side,we use the freeze-thaw method on PBS and we suggest you do the same.
Regarding EGTA and EDTA,they can both be used as chelat to eliminate the interfere of metal ions.They can affect the results from BCA method,but the affect to elisa method is not clear so far.
For 1 mM Na3,we think it might be typed wrong,which should be NaN3.Rigt?NaN3 can not be contained in the sample.Or it will be easily to get false negative result.
For the other components in your RIPA Buffer,we are not clear about their affects to the results.We suggest you use the suggested method.If you have prepared well your sample,you can go on try and do pre-testing.If the sample is not prepared well yet,pls use the suggested method.
For further questions,pls contact us.

I have a one question. Is there any difference in composition between biotin ab. diluent and HRP-avidin diluent?

Thanks for your inquiry.
biotin ab. diluent and HRP-avidin diluent are 2 different components
Biotin-antibody Diluent is for dilutions of Biotin-antibody
HRP-avidin Diluent is for dilutions of HRP-avidin.Pls let me know if you have any further questions. Thank you.

If I use biotin-antibody diluent for HRP-avidin, then Is there any important problem?!

May I know why you would use biotin-antibody diluent for HRP-avidin?Usually we do not suggest that.

Target Background

Function
(From Uniprot)

Functions as cell surface receptor for TIMP1 and plays a role in the activation of cellular signaling cascades. Plays a role in the activation of ITGB1 and integrin signaling, leading to the activation of AKT, FAK/PTK2 and MAP kinases. Promotes cell survival, reorganization of the actin cytoskeleton, cell adhesion, spreading and migration, via its role in the activation of AKT and FAK/PTK2. Plays a role in VEGFA signaling via its role in regulating the internalization of KDR/VEGFR2. Plays a role in intracellular vesicular transport processes, and is required for normal trafficking of the PMEL luminal domain that is essential for the development and maturation of melanocytes. Plays a role in the adhesion of leukocytes onto endothelial cells via its role in the regulation of SELP trafficking. May play a role in mast cell degranulation in response to Ms4a2/FceRI stimulation, but not in mast cell degranulation in response to other stimuli.

Gene References into Functions

Signaling focal contacts via CD63 were identified to facilitate cell adhesion, migration and mediate extracellular matrix-physical cues to modulate hematopoietic stem cells and progenitor cells function. PMID: 28566689
Human CD63-GFP expression was controlled under the rat Sox2 promoter (Sox2/human CD63-GFP), and it was expressed in undifferentiated fetal brains. PMID: 29208635
our results identify the CD63-syntenin-1-ALIX complex as a key regulatory component in post-endocytic HPV trafficking. PMID: 27578500
CD63 and exosome expression is altered in scleroderma dermal fibroblasts PMID: 27443953
CD63 is a critical player in LMP1 exosomal trafficking and LMP1-mediated enhancement of exosome production and may play further roles in limiting downstream LMP1 signaling. PMID: 27974566
TIMP1 signaling via CD63 leads to activation of hepatic stellate cells, which create an environment in the liver that increases its susceptibility to pancreatic tumor cells. PMID: 27506299
Concentrations of Cd63 were elevated in gingival crevicular fluid of patients with pre-eclampsia. PMID: 26988336
CD163(+) TAMs in oral premalignant lesions coexpress CD163 and STAT1, suggesting that the TAMs in oral premalignant lesions possess an M1 phenotype in a Th1-dominated micromilieu. PMID: 26242181
Our findings reveal that elevated levels of TIMP-1 impact on neutrophil homeostasis via signaling through CD63. PMID: 26001794
These findings indicate that rhTIMP-1 promotes clonogenic expansion and survival in human progenitors via the activation of the CD63/PI3K/pAkt signaling pathway PMID: 26213230
It was shown that treatment of macrophages with anti-CD63 inhibits CCR5-mediated virus infection in a cell type-specific manner, but that no inhibition of CXCR4-tropic viruses occurs. PMID: 25658293
TM4SF5 interacted with CD151, and caused the internalization of CD63 from the cell surface. PMID: 25033048
Diesel exhaust exposure during exercise induces platelet activation as illustrated by a dose-response increase in the release of CD62P and CD63. PMID: 25297946
Data show that CD63 is a crucial player in the regulation of the tumor cell-intrinsic metastatic potential by affecting cell plasticity. PMID: 25354204
CD63 tetraspanin is a negative driver of epithelial-to-mesenchymal transition in human melanoma cells. PMID: 24940653
these results indicated that high glycosylation of CD63 by RPN2 is implicated in clinical outcomes in breast cancer patients PMID: 24884960
Collectively, these findings indicate that CD63 may support Env-mediated fusion as well as a late (post-integration) step in the HIV-1 replication cycle. PMID: 24507450
Data suggest that TIMP1 (tissue inhibitor of metalloproteinase-1) acts as chemoattractant for neural stem cells (NSC); TIMP1 enhances NSC adhesion, migration, and cytoskeletal reorganization; these effects are dependent on CD63/CD29 (integrin beta1). PMID: 24635319
Timp1 is assembled in a supramolecular complex containing CD63 and beta1-integrins along melanoma genesis and confers anoikis resistance by activating PI3-K signaling pathway. PMID: 23522389
Antigen-induced p38 MAPK phosphorylation in human basophils essentially contributes to CD63 upregulation PMID: 18727065
Loss of CD63 has a similar phenotype to loss of P-selectin itself, thus CD63 is an essential cofactor to P-selectin. PMID: 21803846
shRNA knockdown in B lymphoblastoid cell line results to increased CD4(+) T-cell recognition PMID: 21660937
decreased levels of CD63 were associated with distant and lymph node metastasis status and does play a direct role in human gastric carcinogenesis. PMID: 21521534
C-terminal modifications that retain LMP1 in Golgi compartments preclude assembly within CD63-enriched domains and/or exosomal discharge leading to NF-kappaB overstimulation. PMID: 21527913
These findings suggest that CD63 plays an early post-entry role prior to or at the reverse transcription step. PMID: 21315401
ameloblastin is expressed in osteoblasts and functions as a promoting factor for osteogenic differentiation via a novel pathway through the interaction between CD63 and integrin beta1 PMID: 21149578
Surface expression of the novel CD63 variant is a distinguishing feature of mast cells, which are are stable, multiple-use cells capable of surviving and delivering several consecutive hits PMID: 20337613
this work provides the first evidence of a TIMP-4/CD63 association in astrocytoma tumor cells PMID: 20693981
CD63 expression results from only the anaphylactic degranulation form of histamine release. PMID: 20633031
Serum sCD163 is a homogenous protein covering more than 94% of the CD163 ectodomain including the haptoglobin-hemoglobin -binding region. PMID: 19581020
Results show that AP-3 is absolutely required for the delivery of CD63 to lysosomes via the trans-Golgi network. PMID: 11907283
downregulation of CD63 antigen is associated with breast tumor progression PMID: 12579280
possible role in HIV-1 infections specific for macrophages PMID: 12610138
Post-translational modification of CD63 may be involved in the functional and morphological changes of MHC class II compartments that occur during dendritic cell maturation PMID: 12755696
relationships between the expression levels of CD61, CD63, and PAC-1 on the platelet surface and the incidences of acute rejection and tubular necrosis as well as the recovery of graft function after renal transplantation PMID: 12826159
Upon platelet interaction with fibrinogen, cholesterol accumulated at the tips of filopodia and at the leading edge of spreading cells; cholesterol-rich raft aggregation was accompanied by concentration of c-Src and CD63 in these cell domains PMID: 12871315
The study on CD63 included its chemistry eg. if it had and O-linked carbohydrate that was digested with O-glycanase. PMID: 12974720
CD63 serves as an adaptor protein that links its interaction partners to the endocytic machinery of the cell. PMID: 14660791
results suggest that CD9, CD63, CD81, and CD82 could play a role in modulating the interactions between immature DCs and their environment, slowing their migratory ability. However, only CD63 would intervene in the internalization of complex antigens PMID: 15130945
Constitutive expression of CD63 may indicate that this factor does not play a direct role in thyroid carcinogenesis PMID: 15375577
CD63 represents an activation-induced reinforcing element, whose triggering promotes sustained and efficient T cell activation and expansion. PMID: 15528334
The linkage of CD63 with PI 4-kinase may result in the recruitment of this signaling enzyme to specific membrane locations in the platelet where it influences phosphoinositide-dependent signaling and platelet spreading. PMID: 15711748
This study identifies a trafficking pathway from CD63-positive multivesicular bodies to the bacterial inclusion, a novel interaction that provides essential lipids necessary for maintenance of a productive intracellular infection. PMID: 16410552
CD63 is recruited to already-budded Weibel-Palade bodies by an AP-3-dependent route PMID: 16683915
CD63-syntenin-1 complex is abundant on the plasma membrane PMID: 16908530
CD63 is a cell surface binding partner for TIMP-1, regulating cell survival and polarization via TIMP-1 modulation of tetraspanin/integrin signaling complex. PMID: 16917503
Chronic urticaria serum-induced CD63 expression assay performed on atopic whole blood by means of tricolor flow cytometry could be the most useful tool for identification of a subset of patients with autoimmune chronic urticaria. PMID: 16918509
In conclusion, using well-defined experimental conditions, the measurement of CD203c up-regulation on basophils in response to specific allergens is as reliable as CD63-BAT for the in vitro diagnosis of patients with IgE-mediated allergy. PMID: 17275019
positive correlation between CD63 and erythrocyte sedimentation rate in rheumatoid arthritis PMID: 17279322
Results suggest that CD63 can be a biomarker for predicting the prognosis in early stages of lung adenocarcinoma. PMID: 17350713

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Subcellular Location

Cell membrane; Multi-pass membrane protein. Lysosome membrane; Multi-pass membrane protein. Late endosome membrane; Multi-pass membrane protein. Endosome, multivesicular body. Melanosome. Secreted, extracellular exosome. Cell surface.

Protein Families

Tetraspanin (TM4SF) family

Tissue Specificity

Detected in platelets (at protein level). Dysplastic nevi, radial growth phase primary melanomas, hematopoietic cells, tissue macrophages.

Database Links

HGNC: 1692

OMIM: 155740

KEGG: hsa:967

STRING: 9606.ENSP00000257857

UniGene: Hs.445570

Code	CSB-E14107h
Size	96T,5×96T,10×96T
Price	Request a Quote
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Intra-assay Precision (Precision within an assay): CV%<8%

Three samples of known concentration were tested twenty times on one plate to assess.

Inter-assay Precision (Precision between assays): CV%<10%

Three samples of known concentration were tested in twenty assays to assess.