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- Antibody Data
- Antigen structure
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- Product number
- MA5-14816 - Provider product page
- Provider
- Invitrogen Antibodies
- Product name
- Histone H4 Monoclonal Antibody (S.99.5)
- Antibody type
- Monoclonal
- Antigen
- Synthetic peptide
- Description
- It is not recommended to aliquot this antibody.
- Antibody clone number
- S.99.5
- Concentration
- 25 µg/mL
Submitted references Novel detection of post-translational modifications in human monocyte-derived dendritic cells after chronic alcohol exposure: Role of inflammation regulator H4K12ac.
Parira T, Figueroa G, Laverde A, Casteleiro G, Gomez Hernandez ME, Fernandez-Lima F, Agudelo M
Scientific reports 2017 Sep 11;7(1):11236
Scientific reports 2017 Sep 11;7(1):11236
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Supportive validation
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- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Western blot was performed using Anti-Histone H4 Monoclonal Antibody (S.99.5) (Product # MA5-14816) and an 11kDa band corresponding to Histone H4 was observed across the cell lines tested along with an uncharacterized band around 20 kDa (*). Acid extracts (15 µg lysate) of SK-O-V3 (Lane 1), Raji (Lane 2) and HeLa (Lane 3) were electrophoresed using NuPAGE™ 12% Bis-Tris Protein Gel (Product # NP0341BOX). Resolved proteins were then transferred onto a nitrocellulose membrane (Product # IB23001) by iBlot® 2 Dry Blotting System (Product # IB21001). The blot was probed with the primary antibody (1:1000 dilution) and detected by chemiluminescence with Goat anti-Mouse IgG (H+L) Superclonal™ Recombinant Secondary Antibody, HRP (Product # A28177,1:4000 dilution) using the iBright™ FL1500 Imaging System (Product # A44115). Chemiluminescent detection was performed using Novex® ECL Chemiluminescent Substrate Reagent Kit (Product # WP20005).
Supportive validation
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Figure 2 Chronic treatment of MDDCs with 0.2% EtOH increases H4K12ac as measured by Immunoblotting. Western blot was carried out for H4K12ac and H4 in histone extracts from untreated control or 0.2% EtOH treated MDDCs. Panel a- shows representative blot highlighting the protein of interest (full length blots are included in the Supplementary Figure S4 ), where first 3 lanes have histone extracts from three biological replicates of untreated MDDCs and next 3 lanes contain histone extracts from three biological replicates of MDDCs chronically treated with 0.2% EtOH. The H4K12ac band appears at 11 kDa while the H4 band appears at 14 kDa. Panel b - optical density of H4K12ac normalized to H4 when analyzed by ImageJ and represented as % control. MDDCs chronically treated with 0.2% EtOH (367.3 %+- 132.9, p = 0.05) showed a higher expression of H4K12ac in comparison to untreated MDDCs (100 %+- 15.05). Panel c - graphical representation of the optical density (OD) values of the H4 band. 0.2% EtOH treated MDDCs (134.6% +- 25.15) contain increased amount of H4 compared to control (100% +- 17.19). Western blot was carried out with histone extracts for control and 0.2% EtOH treated MDDCs from at least 13 different buffy coats. Statistical differences were calculated using t-test when compared to untreated control.
