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Acetyl-Histone H2A (Lys5) Antibody detects endogenous levels of histone H2A only when acetylated at lysine 5.
1. HOW DID OUR ANTIBODY PERFORM? CUSTOMER REVIEW: Histone H2A (Acetyl Lys5) Polyclonal Antibody (STJ97171)
2. WHAT WAS USED? Primary antibody: STJ97171. Histone H2A (Acetyl Lys5) Polyclonal Antibody. Provider: St John’s Laboratory Dilution ratio: 1:250 Application: Immunofluorescence Materials for Validation: Drosophila 3rd instar imaginal discs
3. WHAT WAS THE PROTOCOL? Fixation: 4% paraformaldehyde in PBS + 0.1% Triton-X for 5 minutes. Blocking: 5% FBS in PBS + 0.4% Triton-X for 45 minutes. Primary antibody probing: primary antibody was added to the solution of blocking buffer, 4¡ãC overnight. Wash: PBS + 0.4% Triton-X wash for 3 times. Secondary antibody probing: secondary antibody (Alexa- conjugated goat anti-rabbit 568) was added to the blocking buffer with 1:500 dilution ratio, for 2 hrs. Wash: 1X PBS + 0.4% Triton-X wash for 3 times. Mounting: DAPI-staining conducted in PBS for 10 minutes, followed by mounting with Prolong Gold Antifade. Visualization: Deltavision fluorescence microscope.
4. WHAT WAS THE PROTOCOL? Fixation: 4% paraformaldehyde in PBS + 0.1% Triton-X for 5 minutes. Blocking: 5% FBS in PBS + 0.4% Triton-X for 30 minutes. No. Antigen Primary antibody Primary antibody dilution ratio Secondary antibody dilution ratio Co-staining Antibody Mounting Medium 1 Diploid imaginal disc cells STJ97171. Histone H2A (Acetyl Lys5) Polyclonal Antibody 1:250 1:500 DAPI Prolong Gold
5. WHAT DID THE CUSTOMER THINK? Testing results were provided by Serafin Colmenares from Lawrence Berkeley National Laboratory. They stated that ¡°H2AK5Ac signal overlaps with complete nucleus, except for DAPI bright heterochromatic region. This is in line with the expected signal for this specific modification.¡±
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