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Mouse Granzyme B ProQuantum Immunoassay Kit

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Technical guide & protocol Tech Support
Mouse Granzyme B ProQuantum Immunoassay Kit
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Mouse Granzyme B ProQuantum Immunoassay Kit
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Mouse Granzyme B ProQuantum Immunoassay Kit

How ProQuantum immunoassays work. (A) Antibody-antigen binding: Antibodies bind to two separate epitopes on the antigen (during a 1-hr incubation), which brings the two conjugated oligonucleotides into close proximity. (B) Ligation and amplification of signal (in a qPCR instrument): DNA ligase and a third splint oligonucleotide connector are added to ligate the two ends of the conjugated oligonucleotides, creating a 100-base DNA template. Once the ligase is inactivated at 95°C, the sample is amplified through 40 cycles of annealing and ext... View More {{ $ctrl.currentElement.advancedVerification.fullName }} validation info. View more
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Mouse Granzyme B ProQuantum Immunoassay Kit
Mouse Granzyme B ProQuantum Immunoassay Kit
Mouse Granzyme B ProQuantum Immunoassay Kit (A44238)
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Product Specifications

Analytical sensitivity

<0.3 pg/mL

Assay range

0.32-25,000 pg/mL

Sample type/volume

Serum
2-5 µL
Plasma
2-5 µL
Supernatant
2-5 µL

Hands-on time

30 min

Time-to-result

2 hr

Homogenous (no wash)

Yes

Instrument

qPCR Instruments

Product size

96 Tests

Contents

Assay Dilution Buffer
Antibody-Conjugate A
Antibody-Conjugate B
Antibody-Conjugate Dilution Buffer
Standard
Master Mix
Ligase

Shipping conditions

Dry ice

Storage

-20°C

Protein name

Granzyme B

Species (tested)

Mouse

Assay kit format

Proximity-based Immunoassay

Label or dye

FAM™

About This Kit

The ProQuantum Mouse Granzyme B Immunoassay Kit is designed to provide highly sensitive quantitative measurements of mouse Granzyme B in small sample volumes. Utilizing proximity-based amplification technology, the assay combines the analyte specificity of high-affinity antibody-antigen binding with the signal detection and amplification capabilities of real-time PCR to achieve a simple yet powerful next-generation protein quantitation platform. A user-friendly workflow combined with intuitive Cloud-based software for analytics enables sample-to-answer in just 2 hours.

• High sensitivity-detect low levels of protein with greater sensitivity than traditional methods

• Broad dynamic range-5 logarithmic units, minimizing sample dilutions to ensure they fall within the range

• Small sample consumption-use 2-5 µL of sample (compared to 75 µL for triplicate wells with other methods)

• Fast, easy workflow-2 hours from sample to answer, with no wash steps

• No proprietary instrument to purchase-runs on any real-time PCR instrument

ProQuantum immunoassays utilize a matched pair of target-specific antibodies, each conjugated to a DNA oligonucleotide. During antibody-analyte binding, the two DNA oligos are brought into close proximity, which allows for ligation of the two strands and subsequent creation of a template strand for amplification. This platform leverages the sensitivity and large dynamic range of Applied Biosystems TaqMan real-time PCR technology (Figure 1).

The assay workflow is fast and easy-2 steps in 2 hours. There are a total of 7 components in each kit (Figure 2). First, mix the antibody-conjugates, dilute the sample, and create the standard curve in a working plate. Then, using a multi-channel pipette, add the antibody-conjugates and sample (or standard) into the wells of a PCR plate and incubate for 1 hour. Combine the master mix and ligase and add to the wells of the PCR plate, then run the plate on any qPCR instrument. After the run is complete, import the results file into the ProQuantum cloud-based software at https://apps.thermofisher.com/apps/proquantum. Using this software, the data can be analyzed easily to obtain protein concentration values. The software allows you to set up standard curves, design plate layouts, set up customized assay instructions, and obtain robust statistical group-wise comparisons.

Target information

Granzyme B is a member of the granzyme serine protease family, and is found in the granules of cytotoxic T cells and NK cells. Granzyme B has been described as CGL1 (cathepsin G-like-1), a serine protease expressed only in cytotoxic T-lymphocytes after cell activation, and CTLA-1 (cytotoxic T lymphocyte-associated serine esterase 1) based on identification of mRNA in various cytotoxic T cells, but not observed in non-cytotoxic lymphoid cells. Granzyme B is crucial for the rapid induction of target cell death by apoptosis, induced by interaction with cytotoxic T cells. The receptor involved in this process has been identified as mannose 6-phosphate receptor which functions as a death receptor for Granzyme B during cytotoxic T cell-induced apoptosis. Granzyme B enters target cells to cleave caspase-3 and initiate the caspase cascade leading to DNA fragmentation and apoptosis. Granzyme B can also act through a mitochondrial apoptosis pathway by cleaving the Bid protein. Granzymes are neutral serine proteases, which are stored in specialized lytic granules of cytotoxic T lymphocytes (CTLs) and in natural killer (NK) cells. A number of granzymes (A to G) have been isolated and cloned from mouse CTLs and NK cells, however in man, fewer have been cloned and identified.

For Research Use Only. Not for use in diagnostic procedures. Not for resale without express authorization.

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Bioinformatics

Gene aliases : AI553453, CCP-1/C11, CCP1, Ctla-1, Ctla1, GZB, Gzmb

Gene ID : (Mouse) 14939

Gene symbol : Gzmb

Protein Aliases : CCP1, CTLA-1, Cytotoxic cell protease 1, Fragmentin-2, Granzyme B(G, H), OTTHUMP00000028189, GranzymeB, Human lymphocyte protein (Hlp), Cytotoxic T lymphocyte associated serine esterase 1

UniProt ID (Mouse) P04187

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SIZE

10 x 96 tests

PRICE

USD 400.00

Cat # CHC1263

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Performance Guarantee

If an Invitrogen™ immunoassay doesn't perform as described on our website or datasheet,we'll replace the product at no cost to you, or provide you with a credit for a future purchase.*

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