This antibody gave a positive signal in the following whole cell lysates: HeLa; HEK293; U2OS; Caco2; HCT116; NIH3T3; SW480.
This antibody gave a positive result when used in the following methanol fixed cell lines: HeLa.
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pH: 7.40 Preservative: 0.02% Sodium azide Constituent: PBS Note: Batches of this product that have a concentration < 1mg/ml may have BSA added as a stabilising agent. If you would like information about the formulation of a specific lot, please contact our scientific support team who will be happy to help.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
Use a concentration of 1 µg/ml. Detects a band of approximately 60 kDa (predicted molecular weight: 60 kDa). Abcam recommends using milk as the blocking agent - 3%
Use with paraformaldehyde or methanol fixed cells.
May have oxidoreductase activity. Regulates p38 MAP kinase activity by mediating stress activation of p38alpha/MAPK14 and specifically regulating MAPK14 signaling. Indirectly promotes phosphorylation of MAPK14 and activation of ATF2. The phosphorylation of MAPK14 requires upstream activity of MAP2K4 and MAP2K6. Recruited on chromatin, recognizes and binds trimethylated 'Lys-36' of histone H3 (H3K36me3).
Belongs to the 3-hydroxyisobutyrate dehydrogenase family. NP60 subfamily. Contains 1 A.T hook DNA-binding domain. Contains 1 PWWP domain.
The A.T hook DNA-binding domain is required for the interaction with MAPK14. The PWWP domain probably mediates the binding to H3K36me3.
ICC/IF image of ab124615 stained HeLa cells. The cells were 100% methanol fixed (5 min) and then incubated in 1%BSA / 10% normal goat serum / 0.3M glycine in 0.1% PBS-Tween for 1h to permeabilise the cells and block non-specific protein-protein interactions. The cells were then incubated with the antibody ab124615 at 5µg/ml overnight at +4°C. The secondary antibody (green) was DyLight® 488 goat anti- rabbit (ab96899) IgG (H+L) used at a 1/250 dilution for 1h. Alexa Fluor® 594 WGA was used to label plasma membranes (red) at a 1/200 dilution for 1h. DAPI was used to stain the cell nuclei (blue) at a concentration of 1.43µM.
Immunocytochemistry/ Immunofluorescence - Anti-GLYR1 antibody (ab124615)Image courtesy of an abreview submitted by Dr. Kirk Mcmanus, Univ. of Manitoba/Cancer Care MICB, Canada.
ab124615 (1/400) staining GLYR1 in assynchronous HeLa cells (green). Cells were fixed in paraformaldehyde, permeabilised in 0.5% Triton X100/ PBS and counterstained with DAPI in order to highlight the nucleus (red). For further experimental details please refer to Abreview.
Secondary Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/10000 dilution Developed using the ECL technique
Performed under reducing conditions.
Predicted band size : 60 kDa Observed band size : 60 kDa Additional bands at : 36 kDa,62 kDa,76 kDa. We are unsure as to the identity of these extra bands.
Exposure time : 4 minutesThis blot was produced using a 4-12% Bis-tris gel under the MOPS buffer system. The gel was run at 200V for 50 minutes before being transferred onto a Nitrocellulose membrane at 30V for 70 minutes. The membrane was then blocked for an hour using 3% Milk before being incubated with ab124615 overnight at 4°C. Antibody binding was detected using an anti-rabbit antibody conjugated to HRP, and visualised using ECL development solution.