Invitrogen™ PARP1 (cleaved Asp214) Monoclonal Antibody (HLNC4), eFluor™ 450, eBioscience™

Catalog No.	Quantity
48-666-842	100 Tests

Catalog No. 48-666-842 Supplier Invitrogen™ Supplier No. 48666842

Description

Mouse Monoclonal Antibody

This HLNC4 monoclonal antibody reacts with human poly (ADP-ribose) polymerase (PARP1). This ubiquitous 116 kDa nuclear enzyme is involved in DNA repair. During apoptosis, active caspases -3, -6 and -7 cleave PARP1 after Asp214, thereby inactivating PARP1 and generating two apoptotic fragments sized 85 kDa and 25 kDa. The HLNC4 antibody specifically recognizes the 85 kDa PARP1 fragment produced after cleavage and does not recognize the full-length 116 kDa protein. The following peptide was used as the immunogen: NH2-GVDEVAKKKSKKEKDC-COOH. Applications Reported: This HLNC4 antibody has been reported for use in intracellular staining followed by flow cytometric analysis. Applications Tested: This HLNC4 antibody has been pre-titrated and tested by intracellular staining and flow cytometric analysis of Jurkat cells using the Foxp3/Transcription Factor Staining Buffer Set (Product # 00-5523-00) and protocol. Please refer to BestProtocols: Protocol B: One step protocol for (nuclear) intracellular proteins located under the Resources Tab online. This can be used at 5 μL (0.06 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 instrumen...

Poly ADP-Ribose Polymerase (PARP) uses nicotinamide adenine dinucleotide (oxidized form) NAD as a substrate to catalyse the transfer of ADP-ribose to a variety of nuclear protein acceptors. Proteolysis of PARP to its stable 85kDa fragment is an early marker of programmed cell death (apoptosis) and is mediated by the caspase CPP32 protein. Cleavage occurs between Adp216 and Gly217, a site in PARP conserved across species.

Specifications

• Antigen: PARP1 (cleaved Asp214)
• Applications: Flow Cytometry
• Classification: Monoclonal
• Clone: HLNC4
• Concentration: 5 μL/Test
• Conjugate: eFluor 450
• Formulation: PBS with BSA and 0.09% sodium azide; pH 7.2
• Gene: PARP1
• Gene Accession No.: P09874
• Gene Alias: 5830444G22Rik; ADP-ribosyltransferase (NAD+; ADP-ribosyltransferase (NAD+) poly (ADP-ribose) polymerase); ADP-ribosyltransferase (NAD+, poly (ADP-ribose) polymerase) 1; ADP-ribosyltransferase (NAD+; poly (ADP-ribose) polymerase); ADP-ribosyltransferase (NAD+; poly (ADP-ribose) polymerase) 1; ADP-ribosyltransferase 1; ADP-ribosyltransferase diphtheria toxin-like 1; ADP-ribosyltransferase NAD(+); Adprp; Adprt; ADPRT 1; ADPRT1; AI893648; ARTD1; C80510; DNA ADP-ribosyltransferase PARP1; msPARP; NAD(+) ADP-ribosyltransferase 1; pADPRT-1; PARP; Parp1; PARP-1; poly (ADP-ribose) polymerase 1; poly (ADP-ribose) polymerase family, member 1; poly (ADP-ribose) polymerase); poly [ADP-ribose] polymerase 1; poly(ADP-ribose) polymerase; poly(ADP-ribose) polymerase 1; poly(ADP-ribose) polymerase PARP-1; poly(ADP-ribose) synthetase; poly(ADP-ribosyl)transferase; poly[ADP-ribose] synthase 1; poly[ADP-ribose] synthetase 1; PPOL; Protein poly-ADP-ribosyltransferase PARP1; sPARP-1; synthetase fragment (AA 67); Unknown (protein for IMAGE:8023736); unnamed protein product
• Gene Symbols: PARP1
• Host Species: Mouse
• Purification Method: Affinity chromatography
• Quantity: 100 Tests
• Regulatory Status: RUO
• Primary or Secondary: Primary
• Gene ID (Entrez): 142
• Target Species: Human
• Content And Storage: 4°C, store in dark, DO NOT FREEZE!
• Product Type: Antibody
• Form: Liquid
• Isotype: IgG1 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.