The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
Use a concentration of 5 µg/ml. Detects a band of approximately 55, 85 kDa (predicted molecular weight: 58 kDa).
Use a concentration of 5 µg/ml.
1/250. Perform heat mediated antigen retrieval before commencing with IHC staining protocol.
Use 1µg for 106 cells. ab170191-Mouse monoclonal IgG2a, is suitable for use as an isotype control with this antibody.
Transcriptional regulator. Recognizes and binds to the DNA sequence 5'-CGCCCCCGC-3'(EGR-site). Activates the transcription of target genes whose products are required for mitogenesis and differentiation.
Belongs to the EGR C2H2-type zinc-finger protein family. Contains 3 C2H2-type zinc fingers.
Egr1 was immunoprecipitated using 0.5mg Hela whole cell extract, 5µg of Rabbit polyclonal to Egr1 and 50µl of protein G magnetic beads (+). No antibody was added to the control (-).
The antibody was incubated under agitation with Protein G beads for 10min, Hela whole cell extract lysate diluted in RIPA buffer was added to each sample and incubated for a further 10min under agitation.
Proteins were eluted by addition of 40µl SDS loading buffer and incubated for 10min at 70°C; 10µl of each sample was separated on a SDS PAGE gel, transferred to a nitrocellulose membrane, blocked with 5% BSA and probed with ab55160.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Egr1 antibody [mAbcam55160] (ab55160)This image is courtesy of an abreview submitted by Antibody Solutions Ltd.
ab55160 (1/250) staining Egr1 in paraffin-embedded Human placenta tissue. Tissue underwent fixation in formaldehyde, peroxidase blocking, protein blocking and heat mediated antigen retrieval. The secondary antibody was goat anti rabbit/mouse conjugated to HRP. For further experimental details please refer to abreview.
Overlay histogram showing HeLa cells stained with ab55160 (red line). The cells were fixed with 80% methanol (5 min) and then permeabilized with 0.1% PBS-Tween for 20 min. The cells were then incubated in 1x PBS / 10% normal goat serum / 0.3M glycine to block non-specific protein-protein interactions followed by the antibody (ab55160, 1µg/1x106 cells) for 30 min at 22ºC. The secondary antibody used was DyLight® 488 goat anti-mouse IgG (H+L) (ab96879) at 1/500 dilution for 30 min at 22ºC. Isotype control antibody (black line) was mouse IgG2a [ICIGG2A] (ab91361, 1µg/1x106 cells) used under the same conditions. Acquisition of >5,000 events was performed.
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