Invitrogen™ Mouse Granzyme B ELISA Kit

Catalog No.	Quantity
50-112-5445	96 Tests

Catalog No. 50-112-5445 Supplier Invitrogen™ Supplier No. BMS6029

Description

ELISA

The Mouse Granzyme B ELISA quantitates Ms Granzyme B in mouse serum, plasma, buffered solution, or cell culture medium. The assay will exclusively recognize both natural and recombinant Ms Granzyme B. Principle of the method The Mouse Granzyme B solid-phase sandwich ELISA (enzyme-linked immunosorbent assay) is designed to measure the amount of the target bound between a matched antibody pair. A target-specific antibody has been pre-coated in the wells of the supplied microplate. Samples, standards, or controls are then added into these wells and bind to the immobilized (capture) antibody. The sandwich is formed by the addition of the second (detector) antibody, a substrate solution is added that reacts with the enzyme-antibody-target complex to produce measurable signal. The intensity of this signal is directly proportional to the concentration of target present in the original specimen. Rigorous validation Each manufactured lot of this ELISA kit is quality tested for criteria such as sensitivity, specificity, precision, and lot-to-lot consistency. See manual for more information on validation.

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.

Specifications

• Accession Number: P04187
• Assay Range: 7.8-500 pg/mL
• Assay Sensitivity: 1.6 pg/mL
• Content And Storage: Aluminum pouch(es) with a Microplate coated with monoclonal antibody to mouse Granzyme B
  Biotin-Conjugate anti-mouse Granzyme B antibody
  Streptavidin-HRP
  Mouse Granzyme B Standard lyophilized, 1000 pg/mL upon reconstitution
  Assay Buffer Concentrate 20x (PBS with 1% Tween 20 and 10% BSA)
  Wash Buffer Concentrate 20x (PBS with 1% Tween 20)
  Sample Diluent
  Substrate Solution (tetramethyl-benzidine)
  Stop Solution (1M Phosphoric acid)
  Blue-Dye
  Green-Dye
  Red-Dye
  Adhesive Films
• Conjugate: HRP
• Immunoassay Kit Format: Sandwich ELISA Kit
• Product Type: ELISA
• Sample Type: Plasma, Serum, Supernatant
• Target Species: Mouse
• For Use With (Application): ELISA
• For Use With (Equipment): Absorbance Microplate Reader
• Gene Alias: AI553453; C11; Cathepsin G-like 1; CCP1; CCP-1/C11; CCPI; CGL1; CGL-1; CSPB; CSP-B; Ctla1; Ctla-1; CTSGL1; cytotoxic cell protease 1; cytotoxic serine protease B; Cytotoxic T lymphocyte associated serine esterase 1; cytotoxic T-lymphocyte proteinase 2; cytotoxic T-lymphocyte-associated serine esterase 1; fragmentin; fragmentin 2; fragmentin-2; GLP I; GLP III; GLP-1; granzyme 2; granzyme B; granzyme B (granzyme 2, cytotoxic T-lymphocyte-associated serine esterase 1); granzyme B(G,H); Granzyme-2; GranzymeB; granzyme-like protein 1; granzyme-like protein I; GRB; GZB; Gzmb; HLP; human lymphocyte protein; Human lymphocyte protein (Hlp); Lymphocyte protease; Natural killer cell protease 1; OTTHUMP00000028189; RNKP-1; SECT; T-cell serine protease 1-3EAI553453; C11; Cathepsin G-like 1; CCP1; CCP-1/C11; CCPI; CGL1; CGL-1; CSPB; CSP-B; Ctla1; Ctla-1; CTSGL1; cytotoxic cell protease 1; cytotoxic serine protease B; Cytotoxic T lymphocyte associated serine esterase 1; cytotoxic T-lymphocyte proteinase 2; cytotoxic T-lymphocyte-associated serine esterase 1; fragmentin; fragmentin 2; fragmentin-2; GLP I; GLP III; GLP-1; granzyme 2; granzyme B; granzyme B (granzyme 2, cytotoxic T-lymphocyte-associated serine esterase 1); granzyme B(G,H); Granzyme-2; GranzymeB; granzyme-like protein 1; granzyme-like protein I; GRB; GZB; Gzmb; HLP; human lymphocyte protein; Human lymphocyte protein (Hlp); Lymphocyte protease; Natural killer cell protease 1; OTTHUMP00000028189; RNKP-1; SECT; T-cell serine protease 1-3E; AI553453; CCP-1/C11; CCP1; Ctla-1; Ctla1; GZB
• Gene ID (Entrez): 14939
• Gene Symbol: GZMB
• Interassay CV: 4.1%
• Intraassay CV: 3.5%
• Kit Contents: Pre-coated 96 well plate, Standard, Sample Diluent, Assay Buffer concentrate, Biotinylated Detection Antibody, SAV-HRP, Wash Buffer, Chromogen, Stop Solution, Adhesive Plate Covers, Standard, Sample Diluent, Assay Buffer concentrate, Biotinylated Detection Antibody, SAV-HRP, Wash Buffer, Chromogen, Stop Solution, Adhesive Plate Covers
• Label or Dye: Biotin
• Protein Aliases: CTLA-1, Cytotoxic cell protease 1, CCP1, Fragmentin-2
• Quantity: 96 Tests
• Regulatory Status: RUO
• Sample Volume: Plasma 50 μL, Serum 50 μL, Supernatant 50 μL
• Storage Requirements: 2-8°C
• Synonym: Granzyme 2, Fragmentin 2, Human lymphocyte protein
• Target: Granzyme B
• Test Time: 1 hr 20 min
• Total Assay Time: 3 hr 30 min

Frequently Asked Questions (FAQs)

Do you have information regarding the normal values detectable with your Platinum ELISA Kit?

You can find expected "normal levels" listed in the Platinum ELISA Kit manual under "Expected Values". Please be aware that these normal values were obtained in our laboratories with a limited number of samples. We strongly suggest that each laboratory perform studies in order to establish normal range for their sample populations.

What if it does not look like the supplied reagent volume is sufficient for running the Platinum ELISA?

In case of small reagent volumes, please spin down vial before use to make sure to all contents are collected in the bottom of the tube.

Can I omit the dilution of my Platinum ELISA samples with assay buffer/sample diluent when performing the assay?

We recommend following the instructions for sample dilutions as provided in the manual. We cannot guarantee optimal performance of the assay if internal assay dilutions as instructed were omitted.

Are shorter or longer incubation times possible when using your Platinum ELISA kit?

Incubation times should be followed exactly as stated in the product manual to ensure optimal Platinum ELISA test performance.

Is it necessary to wash the Platinum ELISA plates before usage?

An initial washing step is not necessary but recommended as accuracy may be increased.

Can I always use the same manual for different Platinum ELISA kit lot numbers?

We recommend that you use the manual included with the Platinum ELISA kit.

Is it possible to use the reagents of other Platinum ELISA kits/lots?

The reagents within the Platinum ELISA kits are not interchangeable between different lots of the same kit or between kits against different analytes.

Do I have to run the Platinum ELISA standards and samples in duplicates?

No, but we strongly recommend that you make at least double determinations of each sample and standard point.

Which samples are suitable for Platinum ELISA Kits?

Most ELISAs are suitable for serum, plasma, cell culture supernatants and other body fluids. Detailed information is given in the product manual for each individual kit.

What do Platinum ELISA Kits contain?

All reagents necessary to perform 1 or 10 plate ELISA assays, including optimized capture-antibody pre-coated plates.

I want to collect my tissue culture supernatant daily for a time-course study and run all of my samples at once by ELISA. How should I store my supernatants for use with the eBioscience Platinum ELISA kits?

The supernatants can be harvested and stored -20 degrees C. Avoid freeze/thaw cycles. We recommend that you briefly centrifuge samples after collection to pellet any dead/floating cells remaining in the supernatant. Transfer to new tube for storage.

What concentrations should I use for my recombinant standard when using the eBioscience Platinum ELISA kits?

Each Ready-Set-Go! ELISA Set comes with its own Certificate of Analysis (CofA). The directions on the CofA should be strictly followed to ensure performance of the reagents. The CofA gives specific instructions on how to dilute each component of your lot specific set. The instructions on the CofA should not be deviated from in order to ensure accurate results.

How do I calculate my samples when using the eBioscience Platinum ELISA kits?

The calculation factor is always stated in the manual.

Is the reconstitution volume of the standards the same for different lot numbers when using the eBioscience Platinum ELISA kits?

For some Instant ELISA kits, the reconstitution volume of the standard and blank wells may differ (these components are lot specific). Please use the manual provided in the kit for specific volumes to use.

Do I have to dilute my samples before adding them to the wells when using the eBioscience Platinum ELISA kits?

In general, you can use undiluted samples with the Instant ELISA. Please refer to the product manual to determine if dilution of the sample is required for your kit of interest. The volume of sample required for the individual kits will also be listed in the product manual.

For the eBioscience Platinum ELISA kits, do I have to add the secondary antibody to the wells?

No, the secondary antibody is included as a component of the lyophilized reagents in the wells.

For the eBioscience Platinum ELISA kits, do I have to add recombinant standard to the wells?

No, the recombinant standard is included as a component of the lyophilized reagents in the wells.

Is it possible to store the reagents from the eBioscience Platinum ELISA Kits other than indicated?

Performance of Platinum ELISA cannot be guaranteed if reagents are not stored as indicated.