TotalSeq™-B0044 anti-human Perforin Antibody

Product Details

Verified Reactivity

Human

Reported Reactivity

Cow

Antibody Type

Monoclonal

Host Species

Mouse

Immunogen

Purified granules from the human lymphoma cell line

Formulation

Phosphate-buffered solution, pH 7.2, containing 0.09% sodium azide and EDTA

Preparation

The antibody was purified by chromatography and conjugated with TotalSeq™-B oligomer under optimal conditions.

Concentration

0.5 mg/mL

Storage & Handling

The antibody solution should be stored undiluted between 2°C and 8°C. Do not freeze.

Application

ICPG - Quality tested

Recommended Usage

Each lot of this antibody is quality control tested by intracellular immunofluorescent staining with flow cytometric analysis and the oligomer sequence is confirmed by sequencing. TotalSeq™-B antibodies are compatible with 10x Genomics Single Cell Gene Expression Solutions.

To maximize performance, it is strongly recommended that the reagent be titrated for each application, and that you centrifuge the antibody dilution before adding to the cells at 14,000xg at 2 - 8°C for 10 minutes. Carefully pipette out the liquid avoiding the bottom of the tube and add to the cell suspension. For Proteogenomics analysis, the suggested starting amount of this reagent for titration is ≤ 0.5 µg per million cells in 100 µL volume. Refer to the corresponding TotalSeq™ protocol for specific staining instructions.

Buyer is solely responsible for determining whether Buyer has all intellectual property rights that are necessary for Buyer's intended uses of the BioLegend TotalSeq™ products. For example, for any technology platform Buyer uses with TotalSeq™, it is Buyer's sole responsibility to determine whether it has all necessary third party intellectual property rights to use that platform and TotalSeq™ with that platform.

Application Notes

Clone dG9 primarily recognizes perforin associated with cytotoxic granules⁹. Additional reported applications (for the relevant formats) include: immunoprecipitation, intracellular flow cytometric analysis and immunofluorescence microscopy^5,7, and immunohistochemical staining of acetone-fixed frozen tissue sections and formalin-fixed paraffin-embedded tissue sections^1,4.

Does not cross-react with mouse¹.

Additional Product Notes

TotalSeq™ reagents are designed to profile protein levels at a single cell level following an optimized protocol similar to the CITE-seq workflow. A compatible single cell device (e.g. 10x Genomics Chromium System and Reagents) and sequencer (e.g. Illumina analyzers) are required. Please contact technical support for more information, or visit biolegend.com/totalseq.

The barcode flanking sequences are GTGACTGGAGTTCAGACGTGTGCTCTTCCGATCTNNNNNNNNNN (PCR handle), and NNNNNNNNNGCTTTAAGGCCGGTCCTAGC*A*A (capture sequence). N represents either randomly selected A, C, G, or T, and * indicates a phosphorothioated bond, to prevent nuclease degradation.

View more applications data for this product in our Scientific Poster Library.

Application References

(PubMed link indicates BioLegend citation)

1. Hameed A, et al. 1992. Am. J. Pathol. 140:1025. (IHC)
2. Schaerli P, et al. 2004. J. Exp. Med. 199:1265.
3. Watanabe N, et al. 1997. Blood 90:3662.
4. Mauad T, et al. 2004. Pediatr. Pulmonol. 38:233. (IHC)
5. Barrat FJ, et al. 1999. P. Natl. Acad. Sci. USA 96:8645. (IF)
6. Chen H, et al. 2005. J. Immunol. 175:591.
7. Bryceson YT, et al. 2007. Blood doi:10.1182/blood-2007-02-074468. (IF)
8. Wood SM, et al. 2009. Blood 114:4117. PubMed
9. Makedonas G, et al. 2010. PLoS Pathog. 6:e1000798.

RRID

AB_2941480 (BioLegend Cat. No. 308133)

Antigen Details

Structure

70 kD

Distribution

CTL, NK (cytoplasmic granules)

Function

Mediates targeted cell lysis

Cell Type

NK cells, T cells

Biology Area

Cell Biology, Immunology, Innate Immunity, Neuroscience

Molecular Family

Cytokines/Chemokines

Antigen References

1. Lieberman J. 2003. Nat. Rev. Immunol. 3:361.
2. Trapani J, et al. 2002. Nat. Rev. Immunol. 2:735.

Gene ID

5551 View all products for this Gene ID

UniProt

View information about Perforin on UniProt.org

Related Protocols

• TotalSeq™-B or -C Cell Surface and Intracellular Cytoplasmic Protein Staining with 10x Chromium Single Cell Gene Expression Flex

Other Formats