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<strong> Figure 1 WB Validation in Human Cells </strong><br>Loading: 15 μg of lysate Antibodies: DEDAF, 2227, 1 μ g/mL , 1 h incubation at RT in 5% NFDM/TBST. Secondary: Goat Anti-Rabbit IgG HRP conjugate at 1:10000 dilution.
<strong>Figure 2 WB Validation in Mouse Tissues</strong><br>Loading: 15 μg of lysate Antibodies: DEDAF, 2227, 1 μ g/mL, 1 h incubation at RT in 5% NFDM/TBST. Secondary: Goat Anti-Rabbit IgG HRP conjugate at 1:10000 dilution.
<strong>Figure 3 WB Validation in Rat Tissues</strong><br>Loading: 15 μg of lysate Antibodies: DEDAF, 2227, 1 μ g/mL, 1 h incubation at RT in 5% NFDM/TBST. Secondary: Goat Anti-Rabbit IgG HRP conjugate at 1:10000 dilution.
<strong>Figure 4 Immunohistochemistry Validation of DEDAF in Human Lung</strong><br> Immunohistochemical analysis of paraffin-embedded human lung tissue using anti-DEDAF antibody (2227) at 2 μg/ml. Tissue was fixed with formaldehyde and blocked with 10% serum for 1 h at RT; antigen retrieval was by heat mediation with a citrate buffer (pH6). Samples were incubated with primary antibody overnight at 4°C. A goat anti-rabbit IgG H&L (HRP) at 1/250 was used as secondary. Counter stained with Hematoxylin.
<strong>Figure 5 Immunohistochemistry Validation of DEDAF in Mouse Brain</strong><br> Immunohistochemical analysis of paraffin-embedded mouse brain tissue using anti-DEDAF antibody (2227) at 2 μg/ml. Tissue was fixed with formaldehyde and blocked with 10% serum for 1 h at RT; antigen retrieval was by heat mediation with a citrate buffer (pH6). Samples were incubated with primary antibody overnight at 4°C. A goat anti-rabbit IgG H&L (HRP) at 1/250 was used as secondary. Counter stained with Hematoxylin.
<strong>Figure 6 Immunohistochemistry Validation of DEDAF in Mouse Lung</strong><br> Immunohistochemical analysis of paraffin-embedded mouse lung tissue using anti-DEDAF antibody (2227) at 2 μg/ml. Tissue was fixed with formaldehyde and blocked with 10% serum for 1 h at RT; antigen retrieval was by heat mediation with a citrate buffer (pH6). Samples were incubated with primary antibody overnight at 4°C. A goat anti-rabbit IgG H&L (HRP) at 1/250 was used as secondary. Counter stained with Hematoxylin.
<strong>Figure 7 Immunohistochemistry Validation of DEDAF in Rat Brain</strong><br> Immunohistochemical analysis of paraffin-embedded rat brain tissue using anti-DEDAF antibody (2227) at 2 μg/ml. Tissue was fixed with formaldehyde and blocked with 10% serum for 1 h at RT; antigen retrieval was by heat mediation with a citrate buffer (pH6). Samples were incubated with primary antibody overnight at 4°C. A goat anti-rabbit IgG H&L (HRP) at 1/250 was used as secondary. Counter stained with Hematoxylin.
<strong>Figure 8 Immunofluorescence Validation of DEDAF in Mouse Liver</strong><br> Immunofluorescent analysis of 4% paraformaldehyde-fixed mouse liver tissue labeling DEDAF with 2227 at 20 μg/mL, followed by goat anti-rabbit IgG secondary antibody at 1/500 dilution (red) and DAPI staining (blue).
<strong>Figure 9 Immunofluorescence Validation of DEDAF in A549 Cells</strong><br> Immunofluorescent analysis of 4% paraformaldehyde-fixed A549 cells labeling DEDAF with 2227 at 20 μg/mL, followed by goat anti-rabbit IgG secondary antibody at 1/500 dilution (red).
Figure 1 WB Validation in Human Cells
Loading: 15 μg of lysate Antibodies: DEDAF, 2227, 1 μ g/mL , 1 h incubation at RT in 5% NFDM/TBST. Secondary: Goat Anti-Rabbit IgG HRP conjugate at 1:10000 dilution.
Trial Size

DEDAF Antibody

CATALOG NUMBER: 2227

Clonality:
Polyclonal
Tested Applications:
ELISA, IHC-P, WB, ICC/IF, IHC-P/IF
Host Species:
Rabbit
Species Reactivity:
Human, Mouse, Rat
Conjugate:
Unconjugated
Specifications
Host Species:
Rabbit
Species Reactivity:
Human, Mouse, Rat
Immunogen:
Anti-DEDAF antibody (2227) was raised against a peptide corresponding to 14 amino acid synthetic peptide near the carboxy terminus of human DEDAF.

The immunogen is located within the last 50 amino acids of DEDAF.
Conjugate:
Unconjugated
Tested Applications:
ELISA, IHC-P, WB, ICC/IF, IHC-P/IF
Application Note:
WB: 0.5-2 μg/mL; IHC-P: 1-2 μg/mL; IHC-P/IF: 10-20 μg/mL; ICC/IF: 10-20 μg/mL.

Antibody validated: Western Blot in human, mouse and rat samples; Immunohistochemistry in human, mouse, and rat samples; Immunofluorescence in human and mouse samples. All other applications and species not yet tested.
Positive Control 1:
Cat. No. 1403 - Mouse Brain Tissue Lysate
Positive Control 2:
Cat. No. 1465 - Rat Kidney Tissue Lysate
Positive Control 3:
Cat. No. 1404 - Mouse Liver Tissue Lysate
Positive Control 4:
Cat. No. 17-003 - A549 Cell Slide
Predicted Molecular Weight:
Predicted: 25 kD

Observed: 28 kD
Purification:
DEDAF Antibody is affinity chromatography purified via peptide column.
Clonality:
Polyclonal
Isotype:
IgG
physical-state:
Liquid
Buffer:
DEDAF Antibody is supplied in PBS containing 0.02% sodium azide.
Concentration:
1 mg/mL
Storage Conditions:
DEDAF antibody can be stored at 4°C up to one year. Antibodies should not be exposed to prolonged high temperatures.
Ncbi Official Symbol:
RYBP
Additional Names:
DEDAF Antibody: AAP1, DEDAF, YEAF1, RING1 and YY1-binding protein, Apoptin-associating protein 1, APAP-1
Protein Accession Number:
AF179286
Protein Gi Number:
5802963
Ncbi Gene Id Number:
23429
User Note:
Optimal dilutions for each application to be determined by the researcher.
Background:
DEDAF Antibody: Apoptosis is related to many diseases and induced by a family of cell death receptors and their ligands. Cell death signals are transduced by death domain (DD) death effector domain (DED), and caspase recruitment domain (CARD) containing molecules. Several molecules including caspases and adaptor FADD contain DEDs. A novel protein that interacts with DED of caspase-8 and 10, and FADD was identified recently and designated DEDAF for DED associated factor. DEDAF is identical to the transcriptional repressor RYBP. DEDAF/RYBP is expressed in multiple tissues and cell lines. DEDAF interacts with FADD and augments the formation of CD95/FADD/capase-8 complexes at the cell membrane, and interacts with DED-containing DNA biding protein (DEDD) in the nucleus indicating it is involved in the regulation of both cytoplasmic and nuclear events of apoptosis.
Background Reference 1:
Zheng L, Schickling O, Peter ME, et al. The death effector domain-associated factor (DEDAF) plays distinct regulatory roles in the nucleus and cytoplasm. J. Biol. Chem. 2001; 276:31945-52.
Background Reference 2:
Garcia E, Marcos-Gutierrez C, del Mar Lorente M, et al. RYBP, a new repressor protein that interacts with components of the mammalian Polycomb complex, and with the transcription factor YY1. EMBO J. 1999;18:3404-18.

FOR RESEARCH USE ONLY

For additional information, visit ProSci’s Terms & Conditions Page.

Disclaimer:
Optimal dilutions/concentrations should be determined by the end user. The information provided is a guideline for product use. This product is for research use only.

CATALOG NUMBER:

2227

List Size:
0.02 mg, 0.1 mg

List Price:

Price range: $99.00 through $445.00

Datasheet
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