Mouse Ly9/SLAMF3/CD229 ELISA Kit PicoKine®

CD229/SLAMF3/Lymphocyte Antigen 9 ELISA kit for Mouse

Mouse Ly9/SLAMF3/CD229 ELISA Kit PicoKine™ (96 Tests). Quantitate Mouse Ly9 in cell culture supernatants, serum and plasma (heparin, EDTA, citrate). Sensitivity: 50pg/ml.

Product Info Summary

SKU: EK2011
Size: 96 wells/kit, with removable strips.
Reactive Species: Mouse
Application: ELISA
Sample Types: cell culture supernatants, serum and plasma (heparin, EDTA, citrate).

Product Name

Mouse Ly9/SLAMF3/CD229 ELISA Kit PicoKine®

View all CD229/SLAMF3/Lymphocyte Antigen 9 ELISA kits

SKU/Catalog Number

EK2011

Size

96 wells/kit, with removable strips.

*Question: How many samples can I assay/run in this kit?

Description

Mouse Ly9/SLAMF3/CD229 ELISA Kit PicoKine™ (96 Tests). Quantitate Mouse Ly9 in cell culture supernatants, serum and plasma (heparin, EDTA, citrate). Sensitivity: 50pg/ml.

Storage & Handling

Store at 4°C for 6 months, at -20°C for 12 months. Avoid multiple freeze-thaw cycles (Ships with gel ice, can store for up to 3 days in room temperature. Freeze upon receiving.)

Cite This Product

Mouse Ly9/SLAMF3/CD229 ELISA Kit PicoKine® (Boster Biological Technology, Pleasanton CA, USA, Catalog # EK2011)

Clonality of Antibodies

See Datasheet for details

Immunogen

Expression system for standard: NS0; Immunogen sequence: K48-R453

Sensitivity

<50 pg/ml

Assay Range

93.7 pg/ml - 6,000 pg/ml

Standard Dilution Instructions

serial dilution instructions image

Add 100ul of sample diluent in well #2-#8. Add 200ul standard stock solution to well #1, and serial dilute for well #2-#7 to make a standard curve row. Leave well #8 as blank See datasheet of EK2011 for more details

Cross-reactivity

There is no detectable cross-reactivity with other relevant proteins.

Reactive Species

EK2011 is reactive to LY9 in Mouse samples

Validated Sample Types

cell culture supernatants, serum and plasma (heparin, EDTA, citrate).

Application Guarantee

EK2011 is guaranteed for ELISA in Mouse by Boster Guarantee

See how Boster Bio validate our ELISA kits: ELISA Validation Information

Background of CD229/SLAMF3/Lymphocyte Antigen 9

T-lymphocyte surface antigen Ly-9 is a protein that in humans is encoded by the LY9 gene. This gene is mapped to 1 H3; 1 79.54 cM. Mature human CD229 consists of a 407 amino acid (aa) extracellular domain (ECD) with two Ig‑like V-set and two Ig‑like truncated C2-set domains. It also shows a 22 aa transmembrane segment, and a 179 aa cytoplasmic domain with two immunoreceptor tyrosine-based switch motifs ITSMs. The ECD of human CD229 shares 57%‑59% aa sequence identity with mouse and rat CD229. Within the first two Ig‑like domains that are common to all SLAM proteins, human CD229 shares 24%‑39% aa sequence identity with human 2B4, BLAME, CD2F-10, CD84, CRACC, NTB-A, and SLAM. Alternate splicing generates two additional isoforms that lack the juxtamembrane Ig‑like domain or short cytoplasmic region. CD229 is expressed on T and B cells, thymocytes, and more weakly on NK cells. Homophilic binding between CD229 molecules is mediated by the N-terminal Ig‑like domain. Human and mouse CD229 exhibit cross-species binding. Antigen stimulation of lymphocytes induces CD229 clustering to sites of T cell - B cell contact. Two tyrosines in the cytoplasmic domain are required for association of CD229 with the adaptor proteins AP-2 (μ2 chain) and GRB-2 and both are required for CD229 internalization.

Kit Components

Catalog Number Description Quantity
EK2011-CAP Anti-Mouse LY9 Pre-coated 96-well strip microplate 1
EK2011-ST Mouse LY9 Standard 2 vials, 10 ng/tube
EK2011-DA Mouse LY9 Biotinylated antibody (100x) 100ul
AR1103 Avidin-Biotin-Peroxidase Complex (100x) 100ul
AR1106-1 Sample Diluent 30ml
AR1106-2 Antibody Diluent 12ml
AR1106-3 Avidin-Biotin-Peroxidase Diluent 12ml
AR1104 Color Developing Reagent (TMB) 10ml
AR1105 Stop Solution 10ml
AR1106-5 Wash Buffer (25x) 20ml
PLA-SEA Adhesive plate sealers 4

*The kit components are not available for individual purchase.

Materials Required But Not Included In Kit

  • Microplate Reader capable of reading absorbance at 450nm.
  • Incubator.
  • Automated plate washer (optional).
  • Pipettes and pipette tips capable of precisely dispensing 0.5 µl through 1 ml volumes of aqueous solutions.
  • Multichannel pipettes are recommended for large amount of samples.
  • Deionized or distilled water.
  • 500ml graduated cylinders.
  • Test tubes for dilution.

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Validation Standard Curve O.D. At 450nm

Concentration (pg/ml)093.7187.5375750150030006000
O.D.0.0590.1080.1440.2140.3940.7041.2882.102

Data Example Images

Recommended Sample Dilution Ratios

According to our internal validation assays using this ELISA kit, to detect CD229/SLAMF3/Lymphocyte Antigen 9, Dilution ratio of 1:1, concentration in serum and plasma is 400-900 pg/ml..

Intra Assay Consistency & Inter Assay Consistency

We measured random samples of Mouse Ly9/SLAMF3/CD229 ELISA Kit PicoKine® within the same batch/lot to ensure the consistency of the kits' performances. ELISA intra assay consistency is measured using wells from the same plate/assay kit. ELISA inter assay consistency is measured using wells from different plates from the same batch production/lot.

Intra-Assay PrecisionInter-Assay Precision
Sample123123
n161616242424
Mean (pg/ml)9862522101196682864
Standard deviation4.0226.88101.666.9046.09183.30
CV (%)4.1%4.3%4.6%5.8%6.9%6.4%

Reproducibility

We ensure reproducibility by testing three samples with differing concentrations of CD229/SLAMF3/Lymphocyte Antigen 9 in ELISA kits from four different production batches/lots.

LotsLot 1 (pg/ml)Lot 2 (pg/ml)Lot 3 (pg/ml)Lot 4 (pg/ml)Mean (pg/ml)Standard DeviationCV (%)
Sample 19810293100983.343.4%
Sample 262565561668464526.744.1%
Sample 322102338234419582212156.387.0%
*number of samples for each test n=16.

Gene/Protein Information For LY9 (Source: Uniprot.Org, NCBI)

Gene Name

LY9

Full Name

T-lymphocyte surface antigen Ly-9

Weight

72.139kDa

Alternative Names

CD229 antigen; CD229; cell-surface molecule Ly-9; hly9; Ly9; lymphocyte antigen 9Cell surface molecule Ly-9; mLY9; SLAMF3; T-lymphocyte surface antigen Ly-9 LY9 CD229, SLAMF3, hly9, mLY9 lymphocyte antigen 9 T-lymphocyte surface antigen Ly-9|SLAM family member 3|cell surface molecule Ly-9|signaling lymphocytic activation molecule 3

*if product is indicated to react with multiple species, protein info is based on the gene entry specified above in "species".

For more info on LY9, check out the LY9 Infographic

LY9 infographic

We have 30,000+ of these available, one for each gene! check them out.

In this infographic you will see the following information for LY9: database IDs, super-family, protein function, synonyms, molecular weight, chromosomal locations, tissues of expression, subcellular locations, post translational modifications, and related diseases, research areas & pathways. If you want to see more information included, or would like to contribute to it and be acknowledged, please contact us [email protected].

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7 Customer Q&As for Mouse Ly9/SLAMF3/CD229 ELISA Kit PicoKine®

Question

Q: which procedure should I follow in order to thaw whole blood sample for CD229 ELISA after freezing?

C. Scott

Verified customer

Asked: 2020-07-06

Answer

A: we do not recommend freezing and thaw whole blood. erythrocytes are fragile and, if frozen and thawed, will undergo hemolysis rendering the samples useless. To keep your blood samples to test CD229 for a later time, you should let the blood clot in glass tubes and collect the serum to freeze for later use.

Boster Scientific Support

Answered: 2020-07-06

Question

Q: can you recommend the dilution ratio of serum samples for detection of CD229 in Mouse serum? I am trying to measure a few analytes and it requires 100ul of diluted samples for each well. We have low serum volumes so we like to dilute as much as possible.

Verified Customer

Verified customer

Asked: 2020-01-31

Answer

A: without knowing the physiological or pathological context of your samples we cannot recommend a dilution ratio without performing a pilot test with your samples. Here is how you can perform a pilot study on your own: perform a serial dilution of your samples on the CD229 ELISA kit to make sure you have a linear ascending curve followed by a plateau, which signifies the samples saturating the detection limit of the kit. Then you can pick the dilution ratios from samples in the linear part of the curve as your experimental dilution ratio.
If you are interested in using our ELISA service, you can also send us your sample and we will take care of everything for you. You can check our service details here: bosterbio.com/services/assay-services/ELISA-testing-service
Since you mentioned you have limited samples, our cost effective multiplex ELISA service would fit perfectly for your needs, where we can generate dozens of data points using as little as 25ul sample volume. Information on this service is also in the above link.

Boster Scientific Support

Answered: 2020-01-31

Question

Q: if the enzyme conjugated CD229 antibodies are mixed with the substrate, will that convert the substrate into the enzymatic reaction product? Or the enzyme function is only activated when the antibody is attached with the CD229 antigen?

Verified Customer

Verified customer

Asked: 2019-12-15

Answer

A: The enzyme is always active. Avoid contaminating the substrate with enzyme prior to the incubation otherwise it compromises the assay with false positive signal.

Boster Scientific Support

Answered: 2019-12-15

Question

Q: Are Boster Bio recombinant proteins and antibodies sterile?

J. Brown

Verified customer

Asked: 2019-11-17

Answer

A: although the vials are bottled using aseptic techniques, heat-treated vials, and sterile stock solutions, they are not considered or guaranteed to be sterile. If sterile material is required for an experiment, the material can be filtered through a 0.2 micron filter designed for use with biological fluids.

Boster Scientific Support

Answered: 2019-11-17

Question

Q: What is the optimal O.D. value for CD229 ELISA kit? I used your CD229 ELISA on serum samples. For my positive control, I received an O.D. value of 0.826, while my negative control received a value of 0.136. I obtained both of these controls from the ELISA kit, where your kit's typical data shows O.D. values much higher than my positive control and your background is lower. My samples O.D. values are around 0.225 and the highest is only 0.357. can I consider these samples contain CD229 even though the O.D. values are not very high?

Verified Customer

Verified customer

Asked: 2019-05-20

Answer

A: The absolute O.D. values may change according to incubation time. The more you incubate the higher the O.D. values are going to be. a point of focus should be is whether your sample O.D. values are statistically significantly higher than your blank values. regarding your assay, you could extend your development time in the substrate incubation step to obtain higher O.D. values, as long as your negative controls' O.D. values are not increasing faster in proportion to your positive controls. typically, a sample with O.D. value 2 standard deviations higher than your negative controls can be considered positive. We calculate the sensitivity of this ELISA kit by converting cutoff O.D. value, calculated as the average of 20 negative controls plus 2 standard deviations of the 20 negative controls, into a concentration. in other words, when we claim this CD229 ELISA kit to have sensitivity of 50pg/ml, that means the minimum amount of CD229 that can be declared/interpreted as positive by the above standard is 50pg/ml.

Boster Scientific Support

Answered: 2019-05-20

Question

Q: how many samples can be assayed in a Picokine® ELISA Kit?

Verified Customer

Verified customer

Asked: 2018-09-18

Answer

A: The Picokine® ELISA Kits will generally run a 7-point standard curve, non-specific binding wells, and 39 samples in duplicate. this may might differ by kit so please refer to each datasheet for details.

Boster Scientific Support

Answered: 2018-09-18

Question

Q: can I use heparin plasma as samples in Mouse CD229 Picokine® ELISA Kit (Catalog # EK2011)?

Z. Li

Verified customer

Asked: 2017-03-13

Answer

A: Chelating agents such as EDTA, Heparin and Citrate can attach metal ions from the functional domain of CD229 causing degradation of its protein structure. CD229 may be denatured as a result and may compromise the assay's measurements. The chilating sites could also be too close to the epitopes required for detection and block the antigen antibody reaction. We have tested the CD229 ELISA, treating samples with a number of anticoagulants and decided that heparin, EDTA or citrate can be used for treatment of blood/plasma samples. Do not use other anticoagulents when collecting samples.

Boster Scientific Support

Answered: 2017-03-13

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