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    products/assay-kits/ecl-substrate-kit-high-sensitivity-ab133406.pdf

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Kits/ Lysates/ Other Tools and Reagents Western Blot Tools and Reagents ECL kits
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ECL Substrate Kit (High Sensitivity) (ab133406)

  • Datasheet
  • SDS
  • Protocol Booklet
Reviews (5)Q&A (4)References (38)

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Western blot comparison 1/25000 anti-Transferrin primary antibody, 1/50000 secondary antibody, 20 second exposure
  • Western Blot 1/25000 primary antibody, 1/5000 secondary antibody, 20 second exposure

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Overview

  • Product name

    ECL Substrate Kit (High Sensitivity)
    See all ECL Substrate kits
  • Product overview

    High Sensitivity ECL Substrate Kit ab133406 is designed for the detection of proteins with 23pg-187ng of protein per band. It uses an enhanced chemiluminescent substrate for western blotting that was developed for film imaging and is also compatible with CCD imaging. The high sensitivity ECL substrate produces a strong signal with very low background. Additionally, the ECL signal is long lasting, allowing repeated exposures without fear of losing data.


    Our ECL kits include our popular ECL Substrate Kit ab65623 and our high sensitivity ECL substrate kits:
    - this kit (High Sensitivity ECL Substrate Kit ab133406) to detect 23pg-187ng of protein per band
    - Very High Sensitivity ECL Substrate Kit ab133408 to detect 4.6pg-4.7ng of protein per band
    - Ultra High Sensitivity ECL Substrate Kit ab133409 to detect 1.2pg-2ng of protein per band 


    Detection ranges in pg and ng stated above should be used for guidance only as detection range is dependent on the molecular weight of a protein.


    This product was previously called Optiblot ECL Detect Kit (23pg-187ng).

  • Notes

    Use:
    - 200ml kit for 2000cm2 membrane
    - 500ml kit for 5000cm2 membrane

    The primary antibody can often be diluted 5 to 10 fold more than usual when using this ECL substrate. A typical primary antibody dilution range using the substrate is 1/5000 - 1/20,000, with a typical secondary antibody dilution range of 1/20,000 - 1/100,000. Some optimisation may be required.

     

  • Tested applications

    Suitable for: WBmore details

Properties

  • Storage instructions

    Store at +4°C. Please refer to protocols.
  • Components 200 ml 20 ml 500 ml
    Luminol/Enhancer Solution 1 x 100ml 1 x 10ml 1 x 250ml
    Peroxide Chemiluminescent Detection Reagent 1 x 100ml 1 x 10ml 1 x 250ml
  • Research areas

    • Kits/ Lysates/ Other
    • Tools and Reagents
    • Western Blot Tools and Reagents
    • ECL kits
    • Kits/ Lysates/ Other
    • Tools and Reagents
    • Western Blot Tools and Reagents
    • Detection
    • ECL

Associated products

  • Related Products

    • Prestained Protein Ladder – Mid-range molecular weight (10 - 180 kDa) (ab116027)
    • Prestained Protein Ladder – Broad molecular weight (10-245 kDa) (ab116028)
    • InstantBlue® Coomassie Protein Stain (ISB1L) (ab119211)
    • Bradford Reagent (ab119216)
    • Low Fluorescence Western Membrane (PVDF) (ab133411)

Applications

The Abpromise guarantee

Our Abpromise guarantee covers the use of ab133406 in the following tested applications.

The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.

Application Abreviews Notes
WB
Use at an assay dependent concentration.
Notes
WB
Use at an assay dependent concentration.

Images

  • Western blot comparison 1/25000 anti-Transferrin primary antibody, 1/50000 secondary antibody, 20 second exposure
    Western blot comparison 1/25000 anti-Transferrin primary antibody, 1/50000 secondary antibody, 20 second exposure

    Anti-Transferrin antibody (ab1223) at 1/25000 dilution. Lanes 1-14 Transferrin protein (ab91435), Loading dilution (Lanes 1-8): 12.5, 6.2, 3.1, 1.6, 0.8, 0.3, 0.19µg protein.

     

    Secondary

    HRP conjugated polyclonal to Rabbit IgG (ab97080) at 1/50000 developed using Optiblot ECL Detect and Vendor A ECL subtrate.

    Exposure time: 20 seconds

  • Western Blot 1/25000 primary antibody, 1/5000 secondary antibody, 20 second exposure
    Western Blot 1/25000 primary antibody, 1/5000 secondary antibody, 20 second exposure

    Each lane contains the following amount of COX2 recombinant protein (ab58868): 1) 100ng 2) 50ng 3) 25ng 4) 12.5ng 5) 6.25ng 6) 3.13ng 7) 1.56ng 8) 780pg 9) 390pg 10) 195pg 11) 98pg. The blot was probed with rabbit anti-COX2 antibody (ab15191) at 1/25000 dilution and with a rabbit secondary (ab97080) at 1/5000 dilution. The blot was developed with 20ml of Optiblot ECL Detect.

    Exposure time: 20 seconds

Protocols

  • Protocol Booklet

Click here to view the general protocols

Datasheets and documents

  • SDS download

  • Datasheet download

    Download

References (38)

Publishing research using ab133406? Please let us know so that we can cite the reference in this datasheet.

ab133406 has been referenced in 38 publications.

  • Bo-Yin Z  et al. Unlocking the Recovery Potential: JMJD3 Inhibition-Mediated SAPK/JNK Signaling Inactivation Supports Endogenous Oligodendrocyte-Lineage Commitment Post Mammalian Spinal Cord Injury. Neurochem Res 46:792-803 (2021). PubMed: 33428096
  • Lee-Huang S  et al. Endothelial Nitric Oxide Synthase Knockdown in Human Stem Cells Impacts Mitochondrial Biogenesis and Adipogenesis: Live-Cell Real-Time Fluorescence Imaging. J Clin Med 10:N/A (2021). PubMed: 33562279
  • Tian Y  et al. TRPM2-AS Promotes Bladder Cancer by Targeting miR-22-3p and Regulating GINS2 mRNA Expression. Onco Targets Ther 14:1219-1237 (2021). PubMed: 33658791
  • Wang G  et al. FAM83A and FAM83A-AS1 both play oncogenic roles in lung adenocarcinoma. Oncol Lett 21:297 (2021). PubMed: 33732373
  • Wang K  et al. Vertebral-specific activation of the CX3CL1/ICAM-1 signaling network mediates non-small-cell lung cancer spinal metastasis by engaging tumor cell-vertebral bone marrow endothelial cell interactions. Theranostics 11:4770-4789 (2021). PubMed: 33754027
View all Publications for this product

Customer reviews and Q&As

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1-9 of 9 Abreviews or Q&A

Abreview for ab133406 - ECL Substrate Kit (High Sensitivity)

Excellent Excellent 5/5 (Ease of Use)
Abreviews
Abreviews
abreview image
Super high sensitivity, clear bands were detected in 10 seconds exposure!
Sample: Rat Tissue lysate - whole (cardiac muscle)
Loading amount: 20 µg
Gel Running Conditions: Non-reduced Denaturing
The reviewer received a reward from Abcam’s Loyalty Program in thanks for submitting this Abreview and for helping the scientific community make better-informed decisions.

Abcam user community

Verified customer

Submitted Mar 28 2022

abreview for ab133406 - ECL Substrate Kit (High Sensitivity)

Excellent Excellent 5/5 (Ease of Use)
Abreviews
Abreviews
ECL Substrate Kit (High Sensitivity) provides the user perfect performance when compared to other ECL substrates products. As the product's name suggests, “High Sensitivity” enables the detection from 23pg-187ng of protein per band, which achieve by the enhanced chemiluminescent technic.
The reviewer received a reward from Abcam’s Loyalty Program in thanks for submitting this Abreview and for helping the scientific community make better-informed decisions.

Abcam user community

Verified customer

Submitted Jan 24 2022

ECL kit works on abundant proteins perfectly fine

Excellent Excellent 5/5 (Ease of Use)
Abreviews
Abreviews
We've tried this product on loading control proteins (beta-actin, beta-tubulin, and GAPDH) and it has worked perfectly fine. It has even worked on SOME proteins that are far less abundant such as membrane-bound progesterone receptors in rat brain tissue lysate.
The reviewer received a reward from Abcam’s Loyalty Program in thanks for submitting this Abreview and for helping the scientific community make better-informed decisions.

Collin Gagne

Verified customer

Submitted Oct 14 2019

Optiblot ECL Detect Kit for developing western blots.

Good Good 4/5 (Ease of Use)
Abreviews
Abreviews
abreview image
The solution is highly sensitive, generating signal very fast (beta actin came in 20-30 sec, using the optiblot ECL detection solution developed in chem doc instrument).
The reviewer received a reward from Abcam’s Loyalty Program in thanks for submitting this Abreview and for helping the scientific community make better-informed decisions.

Abcam user community

Verified customer

Submitted Nov 09 2015

Visualization of Western blots

Good Good 4/5 (Ease of Use)
Abreviews
Abreviews
abreview image
This abcam optiblot detect kit gives signal very fast like that of the west dura. For my protein of interest, Signal appeared between 50sec to 1.3mins in chemidoc, B-actin comes even faster (30-40sec). The concentration of the total protein loaded was 40ug).
The reviewer received a reward from Abcam’s Loyalty Program in thanks for submitting this Abreview and for helping the scientific community make better-informed decisions.

Abcam user community

Verified customer

Submitted Aug 07 2015

Question

Can any of Abcam's ECL HRP subtrates be used for ELISA?

Read More

Abcam community

Verified customer

Asked on Aug 06 2013

Answer

ab133409 has been used for ELISA applications.

However, since there are many different readers, plates, etc., testing/evaluation is necessary to assess utility for your specific experimental conditions.

Read More

Jeremy Kasanov

Abcam Scientific Support

Answered on Aug 06 2013

Question

Inquiry: is it suitable to use with AP conjugated secondary antibody ?

Read More

Abcam community

Verified customer

Asked on Jan 18 2013

Answer

Thank you for your enquiry.

I am sorry to confirm that ab133406 is not suitable for detection of AP. This detection system has been designed for optimal detection of HRP, and will not be as sensitive for detection of AP.

I am sorry I am not sure which application you are using, but the following two detection substrate kits are suitable for western blotting, and should also be suitable for IHC:

ab7413 Alkaline Phosphatase chromogen (BCIP/TNBT) https://www.abcam.com/Alkaline-Phosphatase-chromogen-BCIPTNBT-ab7413.html https://www.abcam.com/Alkaline-Phosphatase-chromogen-BCIPTNBT-ab7413.html.

ab7468 Alkaline Phosphatase chromogen (BCIP/NBT) https://www.abcam.com/NBT--BCIP-Solution-Ready-to-Use-Alkaline-Phosphatase-chromogen-ab7468.html https://www.abcam.com/NBT--BCIP-Solution-Ready-to-Use-Alkaline-Phosphatase-chromogen-ab7468.html.

I can recommend to review the online datasheets carefully for further information.

I hope this information will be helpful to you. If you have any further questions, please do not hesitate to contact us.

Read More

Abcam Scientific Support

Answered on Jan 18 2013

Question

Pl advice the alternate to

Super Signal West-Pico Chemiluminiscent Substrate

Read More

Abcam community

Verified customer

Asked on Dec 27 2012

Answer

Thank you for contacting us.

The alternative we have is ab133406; https://www.abcam.com/Optiblot-ECL-Detect-Kit-23pg-187ng-ab133406.html

I hope this information is helpful to you. Please do not hesitate to contact us if you need any more advice or information.

Read More

Abcam Scientific Support

Answered on Dec 27 2012

Question

https://www.abcam.com/Optiblot-Fluorescent-Western-Blot-Kit-ab133410.html
can I quote this against
Roche BM chemiluminesence western blotting Kit (mouse/rabbit)- cat # 11520709001
Regards

Read More

Abcam community

Verified customer

Asked on Dec 10 2012

Answer

Thank you for contacting us. I hope that you are well.

The ab133410 is a fluorescence based kit however the Roche kit cat # 11520709001 is a chemiluminescence based so this will not be a right alternative.

Please suggest ab133406, https://www.abcam.com/Optiblot-ECL-Detect-Kit-23pg-187ng-ab133406.html

This kit is vigorously tested and is fully guaranteed. Other alternatives are ab65623, ab65628, ab133408 and ab133409.

Please note that the kits ab133406 does not have secondary antibody as the Roche kit. However all HRP conjugated secondary would be suitable.

I hope this information is helpful to you. Please do not hesitate to contact us if you need any more advice or information.

Free Rabbit monoclonal antibody with any purchase of a primary antibody, while stocks last! Quote “RABMAB-XBSMG” in your next primary antibody order. For more information, visit the following link: https://www.abcam.com/index.html?pageconfig=resource&rid=15447

Read More

Abcam Scientific Support

Answered on Dec 10 2012

Please note: All products are "FOR RESEARCH USE ONLY. NOT FOR USE IN DIAGNOSTIC PROCEDURES"
For licensing inquiries, please contact partnerships@abcam.com

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