AvivaBlot Femto ECL Reagent (OKCF00002)

Catalog No: OKCF00002

Size:250 mL

AvivaBlot™ Femto Competitor Analysis. Hela whole cell lysates were 2 fold serially diluted from 5 ug/lane to 0.156 ug/lane Total Protein. An Anti-HDAC1 antibody was used at 1:5000, with a Goat anti-Rabbit IgG HRP used at 1:75,000. Exposure time of 180 seconds on an GE Imagequant LAS 4000 was used for image acquisition.

AvivaBlot™ Femto Signal Duration. Quadruplicate blots were prepared with 2 fold dilutions (from 5 ug to 156 ng) of HeLa whole cell lysate were incubated with Rabbit Anti-Human HDAC1 at 1:5000 dilution, and Goat Anti-Rabbit IgG HRP at 1:75,000. Images were acquired at various timepoints up to 20 hrs post substrate addition.

AvivaBlot™ Femto Signal Duration Quantitation. Hela lysate (2.5 ug band #2) HDAC-1 immunoblot band was quantified using an exposure time of 180 seconds per time point over 20 hrs. Relative percentage of signals are reflected, with 5 ug band representing 100%. Values shown for each blot used for competitor analysis.

AvivaBlot™ Femto Signal to Noise Analysis. Triplicate blots for each substrate were prepared using 2 fold dilution of HeLa whole cell lysate from 5 ug to 16 ng. Anti-HDAC1 was used at 1:5000 and Goat Anti-Rabbit IgG HRP at 1:75000 with signals imaged for 180s using a GE Imagequant LAS 4000. Values from background were subtracted from light units from signal and plotted for S/N relative values.

AvivaBlot™ Femto Precision. Western blotting detection of HDAC-1 in HeLa cell lysates with Avivablot Femto. A. Triplicate blots containing 2 fold dilutions of whole cell lysates were incubated with anti-HDAC1 (Human) at 1:5,000, and Goat anti-Rabbit-IgG HRP at 1:75,0000. Blots were imaged for 180 seconds with ImageQuant LAS 40000 (GE Healthcare). B. Integrated averaged signal intensity, showing linear response with amount of lysate over a wide range (DR = Dynamic Range, R2 = linearity).

Product Info

• Product Format: Liquid
• Application: WB
• Additional Information:
  1. Avivablot Femto Part A (5 mL or 125 mL), amber bottle, store at Room Temperature
  2. Avivablot Femto Part B (5 mL or 125 mL), store at Room Temperature
  Mix 1:1 volume/volume prior to use
• Storage: Store at Room Temperature
• Protocol Information:
  1. Perform electrophoresis, membrane transfer and antibody incubation and washes.
  2. Mix Femto Part A and Femto Part B 1:1 volume:volume in a conical tube thoroughly. 7 mLs final is required for a standard mini blot (~64 cm2 ). Working solutions can be stored at 4° C for up to 3 days but should be warmed to room temperature before use.
  3. Apply Femto working solution at room temperature directly to semi-wet blot using 1 mL per 10 cm3 of membrane.
  4. Incubate 2 minutes at RT.
  5. Expose membrane using chemiluminescent imager or x-ray film. Use plastic wrap to avoid wetting film where appropriate.
  6. Exposure time is empirically determined, typical exposures require 60 to 180 seconds.
• Dilution: Primary: 1:1.000 - 1:15.000 Secondary: 1:25.000 - 1:150.000 Varies with chosen antibodies, dilutions from 1 mg/mL stock
• Intended Use: Chemiluminescent substrate for Horseradish Peroxidase based Western Blot detection.
• Description:

  AvivaBlot™ Femto ECL Reagent is a two component enhanced chemiluminescent detection reagent, formulated pre-mixed and ready to use for immunoblot applications. Available as sample size (2 X 5 mL) for a single application (100 cm2) or as 2 X 125 mL bottles or Part A and Part B reagent for 2500 cm2, sufficient for approximately 35 mini gel applications. The signal duration is >24 hours, and the reagent can be stored at room temperature for up to 1 year from date of receipt.

Target Info

• Alias Symbols: OKCF00002, immunoblot, western blot, blot, ECL, HRP, substrate, "Aviva Blot"