Key Specifications Table
| Species Reactivity | Key Applications | Host | Format |
|---|---|---|---|
| M, H | WB, IP, DB | E. coli | Purified |
| Description | |
|---|---|
| Catalogue Number | MABE1016 |
| Description | Anti-pan-ADP-ribose binding reagent |
| Alternate Names |
|
| Background Information | Cat. No.MABE1016, Anti-pan-ADP-ribose binding reagent, is a His-tagged recombinant protein fused to rabbit Fc tag, expressed in and purified from Rosetta™(DE3)pLysS strain of E. coli (Cat. No. 70956). Anti-pan-ADP-ribose binding reagent is useful for the affinity detection of both mono- and poly-ADP-ribosylated proteins on membranes in a manner similar to antibody-based Western and DOT blot analysis, The rabbit Fc tag allows visualization of the binding with conjugated anti-rabbit secondary antibodies. The Fc tag also allows Anti-pan-ADP-ribose binding reagent to be captured on Protein A resins for affinity pull-down applications. |
| Product Information | |
|---|---|
| Format | Purified |
| Presentation | Purified from E. coli by Ni-NTA agarose. Supplied in buffer containing 10 mm Tris pH 7.5, 0.2 M NaCl, 10% Glycerol, 10 mm Imidazole, 1 mm PMSF, 1 mm β-Mercaptoethanol, 10% glycerol without preservatives. |
| Quality Level | MQ100 |
| Applications | |
|---|---|
| Application | Anti-poly-ADP-ribose binding reagent is a reagent that selectively binds to both mono- and poly- ADP ribose for use in Western Blotting, Immunocytochemistry and DOT Blot. |
| Key Applications |
|
| Application Notes | DOT Blot Specificity Analysis: This reagent detected mono(ADPR) on recombinant PARP3 protein, as well as mono(ADPR) and poly(ADPR) on recombinant PARP1 recombinant protein (Lee Kraus, University of Texas Southwestern Medical Center). Immunoprecipitation Analysis: A representative lot of Anti-pan-ADP-ribose binding reagent immunoprecipitated ADP-ribosylated proteins from nuclear extract (Lee Kraus, University of Texas Southwestern). Western Blotting Analysis: A representative lot detected auto-ADP-ribosylation activity of PARP1/2/3 and mutants in the presence of NAD+ or various NAD+ analogs (Gibson, B.A., et Al. (2016). Science. 353(6294):45-50). Western Blotting Analysis: A representative lot detected PARP1-catalyzed NELF-E ADP-ribosylation in cell-free enzymatic reactions as well as ADP-ribosylation of exogenously expressed FLAG-tagged NELF-E in HEK293T cells. PARP inhibitor PJ34 or P-TEFb/CDK9 inhibitor flavopiridol treatment decreased cellular NELF-E ADP-ribosylation level (Gibson, B.A., et Al. (2016). Science. 353(6294):45-50). |
| Biological Information | |
|---|---|
| Concentration | Please refer to lot specific datasheet. |
| Host | E. coli |
| Specificity | poly(ADP-ribose) and mono(ADP-ribose) |
| Species Reactivity |
|
| Species Reactivity Note | Mouse, Human. Predicted to react with all species. |
| Purification Method | Ni-NTA agarose |
| Molecular Weight | Variable depending on the target proteins and the extend of ADP-ribosylation. |
| Product Usage Statements | |
|---|---|
| Quality Assurance | Evaluated by Western Blotting on ADP-ribosylated PARP1 and PARP3 recombinant proteins. Western Blotting Analysis: This reagent detected mono(ADPR) on recombinant PARP3 protein, as well as mono(ADPR) and poly(ADPR) on recombinant PARP1 protein (Lee Kraus, University of Texas Southwestern Medical Center). |
| Usage Statement |
|
| Storage and Shipping Information | |
|---|---|
| Storage Conditions | Stable for 1 year at -80°C from date of receipt. Handling Recommendations: Upon receipt and prior to removing the cap, centrifuge the vial and gently mix the solution. Aliquot into microcentrifuge tubes and store at -80°C. Avoid repeated freeze/thaw cycles, which may damage IgG and affect product performance. |
| Packaging Information | |
|---|---|
| Material Size | 200 µL |