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<strong>Figure 1 Western Blot Validation in Human 293 Cell Lysate</strong><br>Loading: 15 μg of lysate per lane.Antibodies: DRAM 4033 (1 μg/mL), 1h incubation at RT in 5% NFDM/TBST.Secondary: Goat anti-rabbit IgG HRP conjugate at 1:10000 dilution.
<strong>Figure 2 Independent Antibody Validation (IAV) via Protein Expression Profile in Cell Lines</strong><br>Loading: 15 μg of lysates per lane.Antibodies: DRAM 4033 (0.5 μg/mL), DRAM 4035 (2 μg/mL), beta-actin (1 μg/mL) and GAPDH (0.02 μg/mL), 1h incubation at RT in 5% NFDM/TBST.Secondary: Goat anti-rabbit IgG HRP conjugate at 1:10000 dilution.
<strong>Figure 3 Immunofluorescence Validation of DRAM in Human Liver Tissue</strong><br>Immunofluorescent analysis of 4% paraformaldehyde-fixed human liver tissue labeling DRAM with 4033 at 20 μg/mL, followed by goat anti-rabbit IgG secondary antibody at 1/500 dilution (red).
<strong>Figure 4 Immunohistochemistry Validation of DRAM in Human Liver Tissue </strong><br> Immunohistochemical analysis of paraffin-embedded human liver tissue using anti-DRAM antibody (4033) at 2.5 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 Induced Expression of DRAM by CQ in Mouse Neural Precursor Cells (NPCs) (Walls et al., 2010) </strong><br>WT and p53-deficient NPC cells were treated with or without 25μM CQ for 24hr. CQ treatment caused the increased expression level in both DRAM dimer (32kD) and monomer (16kD) compared to the untreated controls. WB results show DRAM induction was p53-dependent.
<strong>Figure 6 Regulated Expression Validation of DRAM in Heterozygous SOD2 KO Mice (Mehta et al., 2011) </strong><br>DRAM expression level detected by anti-DRAM antibodies (4033) decreased in striatum of SOD <sup>-/+ </sup> KO mice (fig. d) as compared to WT mice .
Figure 1 Western Blot Validation in Human 293 Cell Lysate
Loading: 15 μg of lysate per lane.Antibodies: DRAM 4033 (1 μg/mL), 1h incubation at RT in 5% NFDM/TBST.Secondary: Goat anti-rabbit IgG HRP conjugate at 1:10000 dilution.
IAV
Trial Size

DRAM Antibody

CATALOG NUMBER: 4033

Clonality:
Polyclonal
Tested Applications:
ELISA, IF, IHC-P, WB
Host Species:
Rabbit
Species Reactivity:
Human, Mouse, Rat
Conjugate:
Unconjugated
Specifications
Host Species:
Rabbit
Species Reactivity:
Human, Mouse, Rat
Immunogen:
Anti-DRAM antibody (4033) was raised against a peptide corresponding to 16 amino acids near the carboxy terminus of human DRAM.
The immunogen is located within amino acids 170-220 of DRAM.
Conjugate:
Unconjugated
Tested Applications:
ELISA, IF, IHC-P, WB
Application Note:
WB: 1 μg/mL; IF: 20 μg/mL; IHC: 2.5 μg/mL.
Antibody validated: Western Blot in human, mouse and rat samples, Immunofluorescence and Immunohistochemistry in human samples. All other applications and species not yet tested.
Positive Control 1:
Cat. No. 1210 - HEK293 Cell Lysate
Positive Control 2:
Cat. No. 1304 - Human Liver Tissue Lysate
Positive Control 3:
Cat. No. 10-201 - Human Liver Tissue Slide
Predicted Molecular Weight:
Predicted: 14kD, 26kD
Observed: 14 kD, 26kD
Validation:

Independent Antibody Validation in Cell lines (Figure 2) shows similar DRAM expression profile in human, mouse and rat cell lines detected by two independent anti-DRAM antibodies that recognize different epitopes, 4033 against C-terminus domain and 4035 against N-terminus domain.  DRAM proteins are detected in the most tested cell lines at different expression levels by the two independent antibodies. 

Induced Expression Validation (Figure 5): DRAM expression detected by anit-DRAM antibodies (4033) was up-regulated by CQ treatment in WT neural precursor cells.

Regulated expression validation (Figure 6): DRAM expression detected by anit-DRAM antibodies (4033) was down-regulated by  in WT neural precursor cells.

Isoforms:
Human DRAM has 2 isoforms, including isoform 1 (238aa, 26kD) and isoform 2 (128aa, 14kD). Mouse DRAM also has 2 isoforms, including isoform 1 (238aa, 26kD) and isoform 2 (132aa, 15kD). Rat DRAM has only one isoform (238aa, 26kD). 4033 can detect human, mouse and rat isoforms.
Purification:
DRAM Antibody is affinity chromatography purified via peptide column.
Clonality:
Polyclonal
Isotype:
IgG
physical-state:
Liquid
Buffer:
DRAM Antibody is supplied in PBS containing 0.02% sodium azide.
Concentration:
1 mg/mL
Storage Conditions:
DRAM antibody can be stored at 4°C for three months and -20°C, stable for up to one year. As with all antibodies care should be taken to avoid repeated freeze thaw cycles. Antibodies should not be exposed to prolonged high temperatures.
Ncbi Official Symbol:
DRAM1
Additional Names:
DRAM Antibody: DRAM, DRAM, DNA damage-regulated autophagy modulator protein 1, Damage-regulated autophagy modulator
Protein Accession Number:
AAH18435
Protein Gi Number:
22450862
Ncbi Gene Id Number:
55332
User Note:
Optimal dilutions for each application to be determined by the researcher.
Background:
DRAM Antibody: Damage-regulated autophagy modulator (DRAM) is a p53 target gene encoding a lysosomal protein that induces autophagy, a process that degrades cytosolic proteins and organelles. It has been suggested that activation of DRAM by p53 is simultaneous to the activation by p53 of one or more proapoptotic genes such as PUMA, Bax, etc., and that the signaling pathways regulated by these genes promote a full cell death response. By itself, DRAM cannot induce apoptosis, but the fact that it is inactivated in certain cancers highlights the importance of DRAM and suggests that autophagy may play a more important role in cancer than initially suspected. At least two different isoforms of DRAM are known to exist.
Background Reference 1:
Crighton, et al. Cell 2006; 126:121-34.
Background Reference 2:
Gozuacik and Kimchi. Oncogene. 2004; 23:2891-906.
Background Reference 3:
Crighton, et al. Autophagy 2007; 3:72-4.
Citation 1:
Walls et al. Lysosome dysfunction triggers Atg7-dependent neural apoptosis. J Biol Chem. 2010;285(14):10497-507. PMID: 20123985
Citation 2:
Mehta et al. Manganese superoxide dismutase deficiency exacerbates ischemic brain damage under hyperglycemic conditions by altering autophagy. Transl Stroke Res. 2011;2(1):42-50. PMID: 21720543

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:

4033

List Size:
0.02 mg, 0.1 mg

List Price:

Price range: $99.00 through $445.00

Datasheet
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