Invitrogen

CARD8 Polyclonal Antibody

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Datasheet

Protocols

Questions & Answers

Tech Support

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CARD8 Antibody in Immunocytochemistry (ICC/IF) — Immunofluorescence analysis of CARD8 was performed using 70% confluent log phase Hep G2 cells. The cells were fixed with 4% paraformaldehyde for 10 minutes, permeabilized with 0.1% Triton™ X-100 for 15 minutes, and blocked with 2% BSA for 1 hour at room temperature. The cells were labeled with CARD8 Polyclonal Antibody (Product # PA5-75635) at 1:100 dilution in 0.1% BSA, incubated at 4 degree celsius overnight and then labeled with Goat anti-Rabbit IgG (H+L) Superclonal™ Recombinant Secondary Antibody, Alexa Fluor™ 488 (Product # A27034, 1:2000 dilution), for 45 minutes at room temperature (Panel a: Green). Nuclei (Panel b:Blue) were stained with ProLong™ Diamond Antifade Mountant with DAPI (Product # P36962). F-actin (Panel c: Red) was stained with Rhodamine Phalloidin (Product # R415, 1:300). Panel d represents the merged image showing cytoplasmic and nuclear localization. Panel e represents control cells with no primary antibody to assess background. The images were captured at 60X magnification.

CARD8 Antibody in Western Blot (WB) — Western blot was performed using CARD8 Polyclonal Antibody (Product # PA5-75635) and a 61 kDa corresponding to CARD8 full length form and 28 kDa corresponding to C-terminal fragment of CARD8 was observed across cell lines and tissues tested with testis showing higher expression when compared to other tissues. Whole cell extracts (30 µg lysate) of Hep G2 (Lane 1), K-562 (Lane 2), MCF7 (Lane 3), Jurkat (Lane 4), HEK-293 (Lane 5) in Fig. a; Mouse Spleen (Lane 1), Mouse Lung (Lane 2), Mouse Placenta (Lane 3), Mouse Testis (Lane 4), Mouse Brain (Lane 5), Mouse Skeletal Muscle (Lane 6), Mouse Heart (Lane 7) In Fig. b were electrophoresed using NuPAGE™ 4-12% Bis-Tris Protein Gel (Product # NP0322BOX). Resolved proteins were then transferred onto a nitrocellulose membrane (Product # IB23001) by iBlot® 2 Dry Blotting System (Product # IB21001). The blot was probed with the primary antibody (1:1000 dilution) and detected by chemiluminescence with Goat anti-Rabbit IgG (H+L) Superclonal™ Recombinant Secondary Antibody, HRP (Product # A27036, 1:20,000 dilution) using the iBright™ FL1500 Imaging System (Product # A44115). Chemiluminescent detection was performed using SuperSignal™ West Pico PLUS Chemiluminescent Substrate (Product # 34580).

Product Details

PA5-75635

Applications

Tested Dilution

Publications

Western Blot (WB)

1:500-1:2,000

Immunocytochemistry (ICC/IF)

1:100

Product Specifications

Species Reactivity

Human, Mouse, Rat

Host/Isotype

Rabbit / IgG

Class

Polyclonal

Type

Antibody

Immunogen

Recombinant full length Human CARD8.

View immunogen

Conjugate

Unconjugated

Form

Liquid

Concentration

1 mg/mL

Purification

Antigen affinity chromatography

Storage buffer

PBS, pH 7.2, with 50% glycerol

Contains

0.02% sodium azide

Storage conditions

Store at 4°C short term. For long term storage, store at -20°C, avoiding freeze/thaw cycles.

RRID

AB_2719363

Product Specific Information

The antibody was affinity-purified from rabbit antiserum by affinity-chromatography using epitope-specific immunogen and the purity is > 95% (by SDS-PAGE).

Target Information

Apoptosis is related to many diseases and development. Cell death signals are transduced by death domain (DD), death effector domain (DED), and caspase recruitment domain (CARD) containing molecules. CARD containing proteins include some caspases, Apaf-1, CARD4, IAPs, RICK, ARC, RAIDD, BCL-10, and ASC. A novel CARD-containing protein was recently identified and designated CARD8. This protein interacts with DRAL, a p53-responsive protein implicated in the induction of apoptosis, and caspase-1 and its related proteins ICEBERG and pesudo-ICE. Although there are conflicting reports on whether CARD8 acts a pro- or anti-apoptotic protein, it has been suggested that it functions as an adaptor molecule regulating caspase-1 and NF-kappa-B activation.

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

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Bioinformatics

Protein Aliases: apoptotic protein NDPP1; CARD 8; CARD inhibitor of NF-kappaB-activating ligands; CARD-inhibitor of NF-kappa-B-activating ligand; CARD8-CT; CARD8-NT; CARDINAL; caspase recruitment domain family, member 8; Caspase recruitment domain-containing protein 8; Caspase recruitment domain-containing protein 8, C-terminus; Caspase recruitment domain-containing protein 8, N-terminus; DKFZp779L0366; KI; TUCAN; Tumor up-regulated CARD-containing antagonist of CASP9; tumor up-regulated CARD-containing antagonist of caspase nine

Gene Aliases: CARD8; CARDINAL; DACAR; DAKAR; KIAA0955; NDPP; NDPP1; TUCAN

UniProt ID: (Human) Q9Y2G2

Entrez Gene ID: (Human) 22900

Function(s)

apoptotic process activation of cysteine-type endopeptidase activity involved in apoptotic process negative regulation of I-kappaB kinase/NF-kappaB signaling positive regulation of cysteine-type endopeptidase activity involved in apoptotic process positive regulation of interleukin-1 beta secretion

Process(es)

protein binding cysteine-type endopeptidase activator activity involved in apoptotic process NACHT domain binding protein homodimerization activity