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Anti-Poly/Mono-ADP Ribose Rabbit Antibody [A22J18]

Catalog No.: F0957

Application: Reactivity: All

Usage Information

Dilution
WB1:1000 IF1:12000 - 1:48000

Application
WB, IF

Reactivity
All

Source
Rabbit

Storage Buffer
PBS, pH 7.2+50% Glycerol+0.05% BSA+0.01% NaN₃

Storage (from the date of receipt)
-20°C (avoid freeze-thaw cycles), 2 years

Exprimental Methods

IF
Experimental Protocol： Specimen Preparation 1. Aspirate liquid, then cover cells to a depth of 2–3 mm with 4% Paraformaldehyde diluted in 1X PBS. NOTE: Paraformaldehyde is toxic, use only in a fume hood. 2. Fix cells for 15 min at room temperature. 3. Aspirate fixative, rinse three times in 1X PBS for 5 min each. 4. Proceed with Immunostaining. Immunostaining 1. Add theblocking buffer and incubate for 60 min at RT. 2. Prepare primary antibody diluent in antibody dilution buffer as recommended . 3. Aspirate blocking solution, apply diluted primary antibody. 4. Incubate overnight at 4°C. 5. Rinse three times in 1X PBS for 5 min each. 6. Incubate specimens in fluorochrome-conjugated secondary antibody diluted in antibody dilution buffer for 1–2 hr at room temperature in the dark. 7. Rinse three times in 1X PBS for 5 min each. 8. Mount slides usingmounting medium with DAPI and cover with coverslips. 9. For best results, allow mountant to cure overnight at room temperature. For long-term storage, store slides flat at 59°C protected from light.

Experimental Protocol：

Specimen Preparation

1. Aspirate liquid, then cover cells to a depth of 2–3 mm with 4% Paraformaldehyde diluted in 1X PBS.

NOTE: Paraformaldehyde is toxic, use only in a fume hood.

2. Fix cells for 15 min at room temperature.

3. Aspirate fixative, rinse three times in 1X PBS for 5 min each.

4. Proceed with Immunostaining.

Immunostaining

1. Add theblocking buffer and incubate for 60 min at RT.

2. Prepare primary antibody diluent in antibody dilution buffer as recommended .

3. Aspirate blocking solution, apply diluted primary antibody.

4. Incubate overnight at 4°C.

5. Rinse three times in 1X PBS for 5 min each.

6. Incubate specimens in fluorochrome-conjugated secondary antibody diluted in antibody dilution buffer for 1–2 hr at room temperature in the dark.

7. Rinse three times in 1X PBS for 5 min each.

8. Mount slides usingmounting medium with DAPI and cover with coverslips.

9. For best results, allow mountant to cure overnight at room temperature. For long-term storage, store slides flat at 59°C protected from light.

Datasheet & SDS

Biological Description

Specificity
Poly/Mono-ADP Ribose Rabbit mAb recognizes endogenous levels of ADP ribosylated proteins and does not cross-react with other post translational modifications.

Clone
A22J18

Synonym(s)
Poly (ADP-Ribose) Polymer,Poly/Mono-ADP Ribose

Background
Poly(ADP-ribose) (PAR) and mono(ADP-ribose) (MAR) are reversible post-translational modifications mediated by ADP-ribosyltransferases, particularly members of the poly(ADP-ribose) polymerase (PARP) family. MAR refers to the covalent addition of a single ADP-ribose unit—transferred from NAD⁺—to specific amino acid residues (commonly aspartate, glutamate, or lysine) on target proteins. In contrast, PAR consists of two or more ADP-ribose units linked through ribose–ribose glycosidic bonds, forming linear or branched chains. Structurally, each ADP-ribose unit contributes ~0.5 kDa, making PAR a large, highly negatively charged and flexible polymer capable of scaffolding diverse protein assemblies. PARP1, the most abundant and catalytically active nuclear PARP, is rapidly activated by DNA strand breaks and stress signals, catalyzing PARylation to recruit and organize DNA repair complexes. MARylation and PARylation play distinct cellular roles—MAR often regulates individual protein activity or interactions, while PAR serves broader functions as a dynamic structural element in processes such as DNA repair, chromatin remodeling, stress granule formation, mitotic spindle assembly, and nucleolar integrity. The balance between synthesis by PARPs and degradation by enzymes like poly(ADP-ribose) glycohydrolase (PARG) and macrodomain hydrolases is critical for maintaining cellular homeostasis.

Background

Poly(ADP-ribose) (PAR) and mono(ADP-ribose) (MAR) are reversible post-translational modifications mediated by ADP-ribosyltransferases, particularly members of the poly(ADP-ribose) polymerase (PARP) family. MAR refers to the covalent addition of a single ADP-ribose unit—transferred from NAD⁺—to specific amino acid residues (commonly aspartate, glutamate, or lysine) on target proteins. In contrast, PAR consists of two or more ADP-ribose units linked through ribose–ribose glycosidic bonds, forming linear or branched chains. Structurally, each ADP-ribose unit contributes ~0.5 kDa, making PAR a large, highly negatively charged and flexible polymer capable of scaffolding diverse protein assemblies. PARP1, the most abundant and catalytically active nuclear PARP, is rapidly activated by DNA strand breaks and stress signals, catalyzing PARylation to recruit and organize DNA repair complexes. MARylation and PARylation play distinct cellular roles—MAR often regulates individual protein activity or interactions, while PAR serves broader functions as a dynamic structural element in processes such as DNA repair, chromatin remodeling, stress granule formation, mitotic spindle assembly, and nucleolar integrity. The balance between synthesis by PARPs and degradation by enzymes like poly(ADP-ribose) glycohydrolase (PARG) and macrodomain hydrolases is critical for maintaining cellular homeostasis.

References
https://pubmed.ncbi.nlm.nih.gov/24914234/

Reference Table for Selecting PVDF Membrane Pore Size Specifications

Protein Size (kDa)	PVDF Transfer Membrane Pore Size (μm)
< 10	0.22
10-20	0.22
20-30	0.45
30-45	0.45
45-70	0.45
80-100	0.45
100-140	0.45
> 140	0.45

Tech Support

Answers to questions you may have can be found in the inhibitor handling instructions. Topics include how to prepare stock solutions, how to store inhibitors, and issues that need special attention for cell-based assays and animal experiments.

Handling Instructions

Tel: +1-832-582-8158 Ext:3
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Protein Size (kDa)	Separating Gel Percentage
4-40	20%
12-45	15%
10-70	12.5%
15-100	10%
25-100	8%
60-210	5%