Lightning-Link HRP Antibody Labeling Kit (701-0010)
Lightning-Link HRP Antibody Labeling Kit [HRP] 1x 100ug Reaction
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Lightning-Link HRP Antibody Labeling Kit [HRP] 1x 100ug Reaction Summary: | | | | Specificity: | Lightning-Link is the easiest and quickest method available for making antibody (protein) conjugates, requiring just 30 seconds hands-on time. The antibody is covalently bonded to the label in a directional and controlled process at near-neutral pH.
Enzyme conjugates (such as antibody-HRP) give a highly sensitive and specific immuno-reagent. Horseradish peroxidase is frequently the label of choice for ELISA, immunoblotting and immunocytochemistry, due to its small size (40kDa) and high turnover rate, producing a measurable result over a short time period.
The amount of antibody used for labeling should be the same as the pack size of LL-HRP, with specific information available on our protocol page. For any new antibody, optimization of the ratio of antibody to dye is often worthwhile.
If your antibody needs to be concentrated, purified, or cleaned up, please look at our antibody clean up kits. |
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| Publications: | Antibody has been mentioned in at least 4 Publications. |
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| | Note: Not all species have been tested for usefulness with this product. Only those species listed have been tested. We cannot make any guarantees about additional reactivities which may or may not occur. |
View Additional Lightning-Link HRP Antibody Labeling Products Lightning-Link HRP Antibody Labeling Kit [HRP] 1x 100ug Reaction Details: | Uses: | This kit contains 1 x 100ug HRP (up to 400ug Ab) |
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| Unit Size: | 1 x 100ug Reaction |
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| Concentration: | Concentration is not relevant for this product. Please see the protocols for proper use of this product. |
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| Conjugated Antibodies: | 3 Conjugated Antibodies |
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Notes: Storage and components
The kit is shipped at ambient temperature in a tamper-evident polypropylene container. Store at -20 degrees Celsius upon receipt.
Summary of features/benefits:
Easy
No solvents
No powders to weigh out
No desalting/dialysis
High efficiency
100% recovery
Freely scalable (custom pack sizes if required)
Kit contents:
Glass vial(s) of Lightning-Link mix (1 or 3 vials, depending on pack size)
1 vial of LL-AP Modifier reagent
1 vial of LL-AP Quencher reagent
Frequently Asked Questions:
Q1. What functional groups do I need on my protein?
Lightning-Link requires amine groups on the molecule to be labeled. Most proteins have lysine and/or alpha-amino groups. All antibodies will have multiple amine functions.
Q2. Can non-antibody molecules be labeled?
Yes. While labeling of antibodies is a major application, the only requirement is that the protein to be labeled has amine functionality.
Q3. Do I need to purify the conjugate?
No. The chemicals used in Lightning-Link are deactivated by the quencher, and the by-products are benign and do not need to be removed.
Learn more about Lightning-Link Conjugation Kits by reading FAQs Packaging: | Storage: | The kit is shipped at ambient temperature in a tamper-evident polypropylene container. Store at -20 °C upon receipt. |
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| Limitations: | This product is for research use only and is not approved for use in humans or in clinical diagnosis. Products are guaranteed for 6 months from date of receipt, except for peptides and proteins which are guaranteed for 3 months. |
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Background: Easy HRP labeling of your antibody with 30 seconds hands-on time. The Lightning-Link conjugation system represents a quantum leap forward in conjugation technology. It allows you to make antibody conjugates with minimal hands-on time less than 30 seconds. Lightning-Link simplifies immunoassay techniques, such as western blotting, ELISA and immunohistochemistry, by eliminating secondary reagents and cutting the number of incubation and wash steps.
Despite its simplicity, Lightning-Link is a very sophisticated conjugation system in which the antibody is directionally coupled to the label (and not to itself) in a controlled fashion, creating high quality conjugates. As the procedure is not interrupted by desalting steps, trial conjugates can be prepared with microgram quantities of protein and then scaled up with ease. Lightning-Link eliminates problems associated with scale up; hands-on time is essentially independent of process scale. Consistency from batch to batch is easy to achieve, and recoveries approach 100%.
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Images (1) Reviews (5) Related Products (6) Working with Lightning-Link HRP Antibody Labeling
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Jump to: Publications, Images, Reviews, Protocols/Procedures, Ask a Question
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Working with Lightning-Link HRP Antibody Labeling
Lightning-Link HRP Antibody Labeling Kit [HRP] 1x 100ug Reaction Images (1) ELISA Titration: Lightning-Link Antibody Labeling Kit [HRP] [701-0010]-A titration of a typical monoclonal antibody prepared using Lightning-Link and a standard method. |
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Reviewed by:
Anonymous 8/16/2010
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Application:
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Elisa
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Sample Tested:
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DECPG:BSA
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Species:
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Bovine
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Sample Concentration:
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Stock Samples
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Dilution Ratio:
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1:5000
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Incubation Time:
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2 hour
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Incubation Temperature:
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25 Celsius
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Incubation Dilution Buffer:
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10mM PBS
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Method:
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TMB Substrate
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Positive Control:
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DECPG:BSA
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Negative Control:
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BSA
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Spec Wavelength:
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450
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Reviewed by:
Anonymous 8/16/2010
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Application:
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Elisa
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Sample Tested:
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Anti-phosphoserine
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Species:
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Bovine
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Sample Concentration:
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stock
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Results:
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Quick and easy conjugation of lighting link HRP to polyclonal antibody. This was used b/c I wanted to do a sandwich Elisa using the same antibodies so one needed to have and HRP conjugated to it. Worked perfect.
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Dilution Ratio:
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1:5000
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Incubation Time:
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1 hour
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Incubation Temperature:
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25 Celsius
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Incubation Dilution Buffer:
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PBS pH 7.4
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Method:
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TMB Substrate
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Positive Control:
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BSA phosphoserine
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Negative Control:
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BSA
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Spec Wavelength:
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450
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Reviewed by:
Anonymous 2/2/2009
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Application:
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Elisa
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Sample Tested:
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Wheat Gluten
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Species:
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Plant
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Species Other:
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Wheat Gluten
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Sample Concentration:
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2ug/ml
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Lot:
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427
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Results:
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It's hard for me to rate the product because we were using it to label a previously untested antibody that we had in such small quantities that we couldn't label it any other way. The antibody actually did not work in the sandwich assay, but did work in an antigen down test, so I think the conjugation worked OK.
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Incubation Time:
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1 Hour
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Incubation Temperature:
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25 Celsius
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Incubation Dilution Buffer:
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PBS/BSA
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Description:
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anti-gluten
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Dilution Ratio:
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1:500 - 1:64000
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Reviewed by:
Anonymous 10/7/2009
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Application:
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Western Blot
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Sample Tested:
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liver tissue
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Sample Loaded:
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20ug
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Lot:
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664
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Results:
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We are doing IP experiment for pulling down only one protein using our primary antibody. We did experiment to compare with your lightening link kit and 2nd Ab\'s detection pattern. We incubated half of samples (each blot of 4 samples) with primeary and secondary ab, on the other hand, samples were incubated with only the lightning conjuated Ab. When you see the blot, left side is primary and secondary Ab used one, and right side is only incubated with lightning link conjuation 1 st Ab. Clearly, secondary Ab detected something else.
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Blocking Buffer:
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non-fat dry milk
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Concentration:
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5
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Blocking Time:
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1h
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Blocking Temperature:
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37 Celsius
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Incubation Time:
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1h
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Incubation Temperature:
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37 Celsius
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Incubation Dilution Buffer:
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5% non-fat dry milk
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Positive Control:
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no IP sample
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Did you observe bands?:
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True
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Specific Bands:
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100 kDa
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Reviewed by:
Anonymous 2/26/2010
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Application:
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Western Blot
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Sample Tested:
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Pancreatic Cancer Cell protein extract
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Sample Loaded:
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30 ug
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Results:
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This was a trial of the labelling system - which appeared to work great!
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Blocking Buffer:
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Milk blotto
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Concentration:
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5
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Blocking Time:
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1 hr
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Blocking Temperature:
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24 Celsius
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Dilution Ratio:
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1/500
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Incubation Time:
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overnight
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Incubation Temperature:
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4 Celsius
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Incubation Dilution Buffer:
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3% BSA/PBS/0.1% Tween 20
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Method:
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ECL
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Exposure Time:
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10 min
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Positive Control:
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HRP labelled 2nd Antibody
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Did you observe bands?:
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True
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Specific Bands:
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57 kDa
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Non-specific Bands:
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37 120 kDa
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Publications (4)Lightning-Link HRP Antibody Labeling Kit [HRP] 1x 100ug Reaction Product Specific:- Fluorescence Linked Immunosorbant Assays Using Microtiter Plates – Journal of Immunological Methods (2008), 336 pp 135-141.
- Targeting Cancer Stem Cells Through L1CAM Suppresses Glioma Growth - Cancer Research (2008), 68 pp 6043-6048.
more... - Development of a Highly Sensitive ELISA for Aldosterone in Mouse Urine: Validation in Physiological Pathophysiological States of Aldosterone Excess Depletion - Steroids (2008).
- Bovine lactoferrin lactoferricin interfere with intracellular trafficking of Herpes simplex virus-1 – Biochimie (2009), 91 (1) pp 160-164.
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