Antibody data
- Antibody Data
- Antigen structure
- References [0]
- Comments [0]
- Validations
- Western blot [1]
- Immunocytochemistry [1]
- Other assay [1]
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Validation data
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- Product number
- PA5-17027 - Provider product page
- Provider
- Invitrogen Antibodies
- Product name
- H4K20me1 Polyclonal Antibody
- Antibody type
- Polyclonal
- Antigen
- Synthetic peptide
- Description
- It is not recommended to aliquot this antibody.
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Supportive validation
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Western blot analysis was performed on acid extracts (20 µg lysate) of HeLa (Lane 1), MDA-MB-231 (Lane 2) and U-2OS (Lane 3). The blot was probed with anti-Methyl-Histone H4 (Lys20) (Product # PA5-17027, 1:1000 dilution) and detected by chemiluminescence using Goat anti-Rabbit IgG (H+L) Superclonal™ Secondary Antibody, HRP conjugate (Product # A27036, 0.25 µg/mL, 1:4000 dilution). A 14 kDa band corresponding to Methyl-Histone H4 (Lys20) was observed across the cell lines tested.
Supportive validation
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Immunofluorescence analysis of Methyl-Histone H4 (Lys20) was performed using 70% confluent log phase HeLa cells. The cells were fixed with 4% paraformaldehyde for 10 minutes, permeabilized with 0.1% Triton™ X-100 for 10 minutes, and blocked with 1% BSA for 1 hour at room temperature. The cells were labeled with Rabbit Polyclonal Antibody (Product # PA5-17027) at 1:250 dilution in 0.1% BSA and incubated overnight at 4 degree and then labeled with Goat anti-Rabbit IgG (H+L) Superclonal™ Secondary Antibody, Alexa Fluor® 488 conjugate (Product # A27034) at a dilution of 1:2000 for 45 minutes at room temperature (Panel a: green). Nuclei (Panel b: blue) were stained with SlowFade® Gold Antifade Mountant with DAPI (Product # S36938). F-actin (Panel c: red) was stained with Rhodamine Phalloidin (Product # R415, 1:300). Panel d represents the merged image showing nuclear localization. Panel e represents control cells with no primary antibody to assess background. The images were captured at 60X magnification.
Supportive validation
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Antibody specificity for modified targets can be established using peptide arrays by quantifying detection of the target protein along with closely related proteins. Peptide array of Histone H4K20me1 using Anti-Methyl-Histone H4 (Lys20) Polyclonal Antibody: An array of the specific peptide and other relevant peptides when tested using Anti-Methyl-Histone H4 (Lys20) Polyclonal Antibody (Product # PA5-17027), showed that the Histone H4K20me1 modification was specifically recognized by the antibody.