CL300

ProtoGlow ECL

$175.00 – $325.00

Get a free sample of ProtoGlow ECL! Call us at (800) 526-3867 for details!

  • Enhanced chemiluminescent substrate
  • Extended signal life
  • Long shelf life/extra reliability
  • Less antibody needed

 

 
SKU: CL-300 Categories: , , Tag:

Description

Catalog Number: CL-300

  • Enhanced chemiluminescent substrate
  • Extended signal life
  • Long shelf life/extra reliability
  • Less antibody needed

National Diagnostics’ ProtoGlow ECL delivers the latest technology developed for enhanced chemiluminescent detection on Western blots. The unique chemistry of the ProtoGlow ECL system increases the sensitivity of Western blots by up to 20 fold. This allows the detection of proteins at the lower abundance and/or the use of higher dilutions of primary and secondary antibodies, economizing on these expensive reagents.

The improved sensitivity of ProtoGlow ECL is demonstrated below. Serial dilutions of transferrin were blocked and probed with HRP labeled antibody, and then subjected to detection using ProtoGlow ECL. ProtoGlow ECL detected down to 5ng of transferrin.

Gel detected with ProtoGlow ECL
Gel electroblotted onto nitrocellulose and probed with HRP-labelled antibody at 1:30,000 dilution. Detection using ProtoGlow ECL. Blots were exposed for 90 seconds. Lane 1) 100 ng; Lane 2) 50 ng; Lane 3) 25 ng; Lane 4) 10 ng; Lane 5) 5 ng.

 

Long-Lasting Signal

ProtoGlow ECL generates a consistent light output for as long as 120 minutes. This ensures results are more reproducible: exposure to exposure, blot to blot. It also allows multiple exposures of a single blot to be taken to optimize the signal: noise ratio.

Gel detected with ProtoGlow ECL
Sequential exposures of serially diluted HRP conjugated anti-transferrin antibody detected with ProtoGlow ECL. Exposures of 60 seconds were taken at 0, 30, 60 and 180 minutes. Dilutions: Lane 1) 1:10,000; Lane 2) 1:20,000; Lane 3) 1:40,000; Lane 4) 1:80,000; Lane 5) 1:160,000.

Economical

ProtoGlow ECL allows the researcher to use fewer antibodies. The extremely enhanced signal from ProtoGlow ECL allows you to use anywhere from 4 to 40 times less antibody per blot while retaining the same detection sensitivity. This in turn allows you to economize on the consumption of expensive antisera.

Get a free sample of ProtoGlow ECL! Call us at (800) 526-3867 for details

Additional information

Weight 1 lbs
Dimensions 7 × 7 × 11 in

Protocol

ProtoGlow ECL fits into any existing Western blot workflow using the same basic protocol as standard ECL kits. Note: The extreme sensitivity of this substrate may require additional dilution of antibody stocks and/or alterations in blocking protocols to avoid excessive background.

BRIEF PROTOCOL

    1. Prepare your protein blot. PVDF or nitrocellulose may be used.
    2. Block membrane for 60 minutes at room temperature. Use BLOTTO, BSA or nonprotein blocking agents.
    3. Incubate blot with primary antibody for 60 minutes at RT with gentle agitation.
    4. Wash blot in PBST or TBST as follows:
      • One brief wash
      • Once at 15 minutes with 1.0 mL/cm2 membrane
      • Three times for 15 minutes with 0.5 mL/cm2 membrane per wash
    5. Incubate blot with secondary antibody for 60 minutes at RT on an oscillating plate or rocker.
    6. Wash blot three times in TBST for 5 minutes each with at least 0.5 mL/cm2 membrane per wash.
    7. Make the ProtoGlow ECL reagent by mixing equal amounts of parts A and B. Provide 0.1-0.2 mL/cm2 membrane and place on blot for 2 minutes.
    8. Drain excess reagent.
    9. Cover damp blot with plastic wrap and place in cassette.
    10. Insert film into cassette and expose. In most cases, 10-60 seconds is sufficient exposure.

Catalog Number: CL-300: View/Download Full Protocol >

Safety Overview

ProtoGlow ECL Reagent A Safety Overview

Safety Summary (see SDS for complete information before using the product):

Appearance and Odor
Clear, colorless solution

EMERGENCY OVERVIEW – IMMEDIATE HAZARD
CAUSES IRRITATION TO SKIN, EYES, AND RESPIRATORY TRACT. HARMFUL IF SWALLOWED OR INHALED.

EMERGENCY OVERVIEW – CHRONIC HAZARD WARNING
CHRONIC DERMATITIS MAY FOLLOW SKIN CONTACT.

ProtoGlow ECL Reagent B Safety Overview

Safety Summary (see SDS for complete information before using the product):

Appearance and Odor
Clear, colorless solution

EMERGENCY OVERVIEW – IMMEDIATE HAZARD
N/A

EMERGENCY OVERVIEW – CHRONIC HAZARD WARNING
N/A

Full SDS

ProtoGlow ECL Reagent A

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ProtoGlow ECL Reagent B

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