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<strong>Figure 1 Immunoflouorescence Validation of AMBRA1 in HeLa Cells</strong><br>Immunofluorescent analysis of PFA-fixed HeLa cells labeling AMBRA1 with 4555 at 20 μg/mL, followed by goat anti-rabbit IgG secondary antibody at 1/1000 dilution (red) and DAPI staining (blue). Alpha tubulin was stained with anti-alpha tubulin antibody following by goat anti-mouse IgG secondary antibody (green). Images were captured with confocal microscopy.
<strong>Figure 2 WB Validation in Human Cell Lines </strong><br>Loading: 15 μg of lysate per lane Antibodies: AMBRA1 4555, 1 μg/mL , 1 h incubation at RT in 5% NFDM/TBST. Secondary: Goat Anti-Rabbit IgG HRP conjugate at 1:10000 dilution.
<strong>Figure 3 Western Blot Validation in 3T3/NIH Cells </strong><br>Loading: 15 μg of lysateAntibodies: AMBRA1 4555, 1 μg/mL, 1h incubation at RT in 5% NFDM/TBST.Secondary: Goat anti-rabbit IgG HRP conjugate at 1:10000 dilution.
<strong>Figure 4 Immunohistochemistry Validation of AMBRA1 in Rat Colon Tissue </strong><br> Immunohistochemical analysis of paraffin-embedded rat colon tissue using anti-AMBRA1 antibody (4555) 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 Immunofluorescence Validation of AMBRA1 in K562 Cells</strong><br>Immunofluorescent analysis of 4% paraformaldehyde-fixed K562 Cells labeling AMBRA1 with 4555 at 20 μg/mL, followed by goat anti-rabbit IgG secondary antibody at 1/500 dilution (red) DAPI staining (blue).
Figure 1 Immunoflouorescence Validation of AMBRA1 in HeLa Cells
Immunofluorescent analysis of PFA-fixed HeLa cells labeling AMBRA1 with 4555 at 20 μg/mL, followed by goat anti-rabbit IgG secondary antibody at 1/1000 dilution (red) and DAPI staining (blue). Alpha tubulin was stained with anti-alpha tubulin antibody following by goat anti-mouse IgG secondary antibody (green). Images were captured with confocal microscopy.
Trial Size

Ambra1 Antibody

CATALOG NUMBER: 4555

Clonality:
Polyclonal
Tested Applications:
ELISA, IHC-P, WB, ICC/IF
Host Species:
Rabbit
Species Reactivity:
Human, Mouse, Rat
Conjugate:
Unconjugated
Specifications
Host Species:
Rabbit
Species Reactivity:
Human, Mouse, Rat
Homology:
Predicted species reactivity based on immunogen sequence: Horse (98%), Pig (97%)
Immunogen:
Ambra1 antibody was raised against a 15 amino acid synthetic peptide from near the carboxy terminus of human Ambra1.
The immunogen is located within the last 50 amino acids of Ambra1.
Conjugate:
Unconjugated
Tested Applications:
ELISA, IHC-P, WB, ICC/IF
Application Note:
WB: 1-2 μg/mL; IHC: 1-5 μg/mL; IF: 20 μg/mL.

Antibody validated: Western Blot in human and mouse samples; Immunocytochemistry and Immunofluorescence in human samples. All other applications and species not yet tested.
Positive Control 1:
Cat. No. 1282 - 3T3 (NIH) Cell Lysate
Predicted Molecular Weight:
Predicted: 143kD

Observed: 143 kD
Purification:
Ambra1 Antibody is affinity chromatography purified via peptide column.
Clonality:
Polyclonal
Isotype:
IgG
physical-state:
Liquid
Buffer:
Ambra1 Antibody is supplied in PBS containing 0.02% sodium azide.
Concentration:
1 mg/mL
Storage Conditions:
Ambra1 Antibody can be stored at 4°C up to one year. Antibodies should not be exposed to prolonged high temperatures
Ncbi Official Symbol:
AMBRA1
Additional Names:
Ambra1 Antibody: DCAF3, WDR94, KIAA1736, Activating molecule in BECN1-regulated autophagy protein 1
Protein Accession Number:
Q9C0C7
Protein Gi Number:
166215833
Ncbi Gene Id Number:
55626
User Note:
Optimal dilutions for each application to be determined by the researcher.
Background:
Ambra1 Antibody: Autophagy, the process of bulk degradation of cellular proteins through an autophagosomic-lysosomal pathway is important for normal growth control and may be defective in tumor cells. It is involved in the preservation of cellular nutrients under starvation conditions as well as the normal turnover of cytosolic components. Beclin-1, a principal regulator of autophagosome formation, is in turn regulated by Ambra1. Ambra1 associates with Beclin-1 through a region near its center as determined by yeast two-hybrid assay. Null mutations in this gene in mice resulted in embryonic lethality with severe neural tube defects associated with autophagy impairment, accumulation of ubiquitinated proteins, unbalanced cell proliferation and excessive apoptotic death. Furthermore, down-regulation of Ambra1 in cultured cells though RNA interference decreased the level of rapamycin- and nutrient starvation-induced autophagy. Multiple isoforms of Ambra1 are known to exist.
Background Reference 1:
Gozuacik D and Kimchi A. Autophagy as a cell death and tumor suppressor mechanism. Oncogene2004; 23:2891-906.
Background Reference 2:
Kisen GO, Tessitore L, Costelli P, et al. Reduced autophagic activity in primary rat hepatocellular carcinoma and ascites hepatoma cells. Carcinogenesis1993; 14:2501-5.
Background Reference 3:
Liang XH, Jackson S, Seaman M, et al. Induction of autophagy and inhibition of tumorigenesis by beclin 1. Nature1999; 402:672-6.
Background Reference 4:
Fimia GM, Stoykova A, Romagnoli A, et al. Ambra1 regulates autophagy and development of the nervous system. Nature2007; 447:1121-5.

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:

4555

List Size:
0.02 mg, 0.1 mg

List Price:

Price range: $99.00 through $445.00

Datasheet
Shipping Info

$65 Standard Overnight flat rate for as many products shipped within the United States.

$35 Dry Ice Shipment fee may be required on some items.

Note: Online orders only accepted for shipment within the USA. Click here for the list of international distributors.

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