Invitrogen™ Perforin Monoclonal Antibody (dG9 (delta G9)), eFluor™ 450, eBioscience™, Invitrogen™

Catalog No.	Quantity
50-112-4242	100 Tests

Catalog No. 50-112-4242 Supplier Invitrogen™ Supplier No. 48999442

Description

Mouse Monoclonal Antibody

Description: The dG9 antibody clone reacts with human perforin (pore-forming protein, pfp). Perforin is one of the cytolytic mediators present in the cytoplasmic granules of cytotoxic T lymphocytes (CTL) and natural killer cells (NK). Perforin is involved in the killing function by CTLs and NKs and has an important role in the immune response against tumors and virus infections. Applications Reported: This dG9 (delta G9) antibody has been reported for use in intracellular staining followed by flow cytometric analysis. Applications Tested: This dG9 (delta G9) antibody has been pre-titrated and tested by intracellular staining followed by flow cytometric analysis of normal human peripheral blood cells using the Intracellular Fixation & Permeabilization Buffer Set (cat. 88-8824) and protocol. Please refer to Best Protocols: Protocol A: Two step protocol for (cytoplasmic) intracellular proteins located under the Resource Tab online. This can be used at 5 μL (0.125 μg) per test. A test is defined as the amount (μg) of antibody that will stain a cell sample in a final volume of 100 μL. Cell number should be determined empirically but can range from 10^5 to 10^8 cells/test. eFluor™ 450 is an alternative to Pacific Blue™. eFluor™ 450 emits at 445 nm and is excited with the Violet laser (405 nm). Please make sure that your instrument is capable of detecting this fluorochome.

Perforin is one of the major cytolytic proteins of cytolytic granules. Perforin is a cytolytic mediator and is stored in and released by cytoplasmic granules. Moreover, perforin is involved in immune defense against tumors and virus infections as mediated by cytotoxic lymphocytes. Perforin is a 555 amino acid protein with a 21 amino acid signal peptide, and has a molecular weight of 70 to 75 kD. Perforin is a pore forming protein with a mechanism of transmembrane channel formation similar to C9, and homology between perforin and C9 have been demonstrated. Studies show that perforin is expressed only in killer cell lines and not in helper T lymphocytes or other tumor cells tested. Perforin is known to be a key effector molecule for T-cell- and natural killer-cell-mediated cytolysis. Defects in the perforin gene cause familial hemophagocytic lymphohistiocytosis type 2 (HPLH2), a rare and lethal autosomal recessive disorder of early childhood. Alternative splicing results in multiple transcript variants of perforin.

Specifications

• Antigen: Perforin
• Applications: Flow Cytometry
• Classification: Monoclonal
• Clone: dG9 (delta G9)
• Concentration: 5 μL/Test
• Conjugate: eFluor 450
• Formulation: PBS with BSA and 0.09% sodium azide; pH 7.2
• Gene: PRF1
• Gene Accession No.: P14222
• Gene Alias: Cyta; Cytolysin; FLH2; HPLH2; lymphocyte pore forming protein; lymphocyte pore-forming protein; OMAK; OTTHUMP00000019759; P1; perforin 1; perforin 1 (pore forming protein); Perforin1; perforin-1; Pfn; PFN1; Pfp; PGFL; PIGF; PIGF-2; PLGF; pore forming protein; pore-forming protein; PRF1; Prf-1; PRF1 (pore forming protein 1); RATCYTA; RP11-710A11.3
• Gene Symbols: PRF1
• Host Species: Mouse
• Purification Method: Affinity chromatography
• Quantity: 100 Tests
• Regulatory Status: RUO
• Primary or Secondary: Primary
• Gene ID (Entrez): 396595, 5551
• Target Species: Human, Pig
• Content And Storage: 4°C, store in dark, DO NOT FREEZE!
• Product Type: Antibody
• Form: Liquid
• Isotype: IgG2b κ

Frequently Asked Questions (FAQs)

I am unable to analyze my cells stained with eFluor Organic Dyes today. What options do I have?

Our options will depend on the samples you are analyzing.

If cell viability is not critical, you can store your stained samples at 4 degrees C or on ice overnight in the dark and analyze the following day.

For samples stained with eFluor organic fluorochromes, we recommend that cells be suspended in 100 uL of Flow Cytometry Staining Buffer (Cat. No. 00-4222) and 100 uL of eBioscience IC Fixation Buffer (Cat. No. 00-8222); samples can be incubated for up to 3 days at 4 degrees C in the dark. Alternatively, the 1-step Fix/Lyse Solution (Cat. No. 00-5333) can be used. This is a great option when working with whole blood but also works for other cell types.

Can the eFluor Organic fluorochromes be used for intracellular staining?

Yes, the eFluor Organic fluorochromes can be used for intracellular staining. The eFluor organic fluorochromes maintain bright signal and require minimal changes in compensation when fixed with eBioscience IC Fixation Buffer (Cat. No. 00-8222-49) and Permeabilization Buffer (Cat. No. 00-8333-56) or 1-step Fix/Lyse Solution (Cat. No. 00-5333-54, 00-5333-57) (as compared to live cells).

What buffers are compatible with the eFluor Organic fluorochromes and eVolve QDots?

The eFluor Organic fluorochromes and eVolve QDots can be used with flow staining buffers containing PBS and protein.

What is the difference between eFluor Organic Dyes and eVolve Qdots?

The eFluor Organic Dyes (eFluor 450, APC-eFluor 780, PerCP-eFluor 710, eFluor 710) are conventional fluorochromes. In contrast, the eVolve line of products are Quantum dots.

Are the eFluor Organic Dyes photo-labile?

As with other fluorochromes, we recommend minimal exposure to light to maintain optimal signal.

For Research Use Only.