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    streptavidin-conjugation-kit-lightning-link-ab102921.pdf

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Streptavidin Conjugation Kit - Lightning-Link® (ab102921)

  • Datasheet
  • SDS
Submit a review Q&A (2)References (33)

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Conjugation - Streptavidin Conjugation Kit - Lightning-Link® (ab102921)
  • Conjugation - Streptavidin Conjugation Kit - Lightning-Link® (ab102921)
  • Conjugation - Streptavidin Conjugation Kit - Lightning-Link® (ab102921)
  • Streptavidin Conjugation Kit

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関連製品

製品の概要

  • 製品名

    Streptavidin Conjugation Kit - Lightning-Link®
  • 製品の概要

    Streptavidin Conjugation Kit / Streptavidin Labeling Kit ab102921 uses a simple and quick process for streptavidin labeling / conjugation of antibodies. It can also be used to conjugate other proteins or peptides. Learn about our antibody labeling kits and their advantages.


    To conjugate an antibody to Streptavidin using this kit:
    - add modifier to antibody and incubate for 3 hrs
    - add quencher and incubate for 30 mins
    The conjugated antibody can be used immediately in WB, ELISA, IHC etc. No further purification is required and 100% of the antibody is recovered for use.


    Learn about buffer compatibility below; for incompatible buffers and low antibody concentrations, use our rapid antibody purification and concentration kits. Use the FAQ to learn more about the technology, or about conjugating other proteins and peptides to Streptavidin.


    Custom size conjugation kits up to 100 mg are available on demand. Please contact us to discuss your requirements.

  • 特記事項

    This product is manufactured by Expedeon, an Abcam company, and was previously called Lightning-Link® Streptavidin Labeling Kit. 708-0005 is the same as the 100 µg size. 708-0010 is the same as the 3 x 100 ug size. 708-0030 is the same as the 3 x 10 ug size. 708-0015 is the same as the 1 mg size.

    Amount and volume of antibody for conjugation to Streptavidin

     Kit size Recommended maximum
    amount of antibody
     
    Maximum antibody 
    volume1
    3 x 10 µg 3 x 10 µg 3 x 10 µL
    100 µg 100 µg  100 µL
    3 x 100 µg 3 x 100 µg  3 x 100 µL
    1 x 1 mg 1 x 1 mg 1 x 1 mL

    1 Ideal antibody concentration is 1mg/ml. 0.5 - 1 mg/ml can be used if the maximum antibody volume is not exceeded. Antibodies > 5mg/ml or < 0.5 mg/ml should be diluted /concentrated.

     

    Buffer Requirements for Conjugation

    Buffer should be pH 6.5-8.5.

    Compatible buffer constituents 
    If a concentration is shown, then the constituent should be no more than the concentration shown. If several constituents are close to the limit of acceptable concentration, then this can inhibit conjugation.

    50mM / 0.6% Tris1 0.1% BSA 50% glycerol
    0.1% sodium azide PBS Potassium phosphate
    Sodium chloride HEPES Sucrose
    Sodium citrate EDTA Trehalose

    1 Tris buffered saline is almost always ≤ 50 mM / 0.6%

    Incompatible buffer constituents

    Thiomerosal Proclin Glycine
    Arginine Glutathione DTT

    If a constituent of the buffer containing your antibody or protein is not listed above, please check the FAQ or contact us.

    Only purified antibodies are suitable for use, ie. where other proteins, peptides, or amino acids are not present: antibodies in ascites fluid, serum or hybridoma culture are not suitable for conjugation.

    Storing and handling conjugation kits

    Lyophilized Lightning-Link® components are hygroscopic.

    Kits are intentionally shipped at ambient temperature with silica gel to avoid exposure to moisture. Upon receipt, store the kit frozen and protect from moisture. Before opening the outer container, allow the lyophilized components to reach room temperature to minimize condensation.

製品の特性

  • 保存方法

    Store at -20°C. Please refer to protocols.
  • 内容 1 mg 100 µg 3 x 10 µg 3 x 100 µg
    ab274106 - Modifier reagent 1 x 200µl 1 x 200µl 1 x 200µl 1 x 200µl
    ab274296 - Quencher reagent 1 x 200µl 1 x 200µl 1 x 200µl 1 x 200µl
    ab274136 - Streptavidin mix 1 x 1mg 1 x 100µg 3 x 10µg 3 x 100µg
  • 研究分野

    • Tags & Cell Markers
    • Epitope Tags
    • Conjugates
    • Kits/ Lysates/ Other
    • Kits
    • Conjugation Kits
    • Streptavidin

関連製品

  • Related Products

    • Antibody Concentration And Clean-Up Kit (ab102778)
    • Antibody Purification Kit (Protein A) (ab102784)

画像

  • Conjugation - Streptavidin Conjugation Kit&nbsp;- Lightning-Link&reg; (ab102921)
    Conjugation - Streptavidin Conjugation Kit - Lightning-Link® (ab102921)Image from Pereira et al., Traffic, 21(5):375-385; doi: 10.1111/tra.12728. Reproduced under the Creative Commons license https://creativecommons.org/licenses/by/4.0/

    Pereira, Pedro M., et al used Streptavidin Conjugation Kit - Lightning-Link® (ab102921) as part of examining super-resolution fluorescent probes for Single-molecule localization microscopy. They used the kit to conjugate Streptavidin to anti-mouse secondary antibody for use in fixed-cell imaging.
    D, Pre-acquisition snapshot (WF) and SMLM reconstruction (STORM) of β-tubulin immunolabelling with B3S9L SB (B3S9L-ATTO555-BHQ2) and control probe (B3S9L-ATTO555). Scale bars are 5 µm. E, Schematic representation of the strategy used to perform β-tubulin immunolabelling (left), measurement of Full With Half Maximum (FWHM) and distances between structures on the highlighted regions in D

  • Conjugation - Streptavidin Conjugation Kit&nbsp;- Lightning-Link&reg; (ab102921)
    Conjugation - Streptavidin Conjugation Kit - Lightning-Link® (ab102921)Image from Hoffmann et al., Sci Rep., 10(1):8869; doi: 10.1038/s41598-020-65860-x.Reproduced under the Creative Commons license https://creativecommons.org/licenses/by/4.0/

    Hoffmann, Ricarda M., et al used Streptavidin Conjugation Kit - Lightning-Link® (ab102921) as part of examining development of anticancer Antibody-Drug Conjugates. They used the kit to conjugate Streptavidin to Trastuzumab and trastuzumab emtansine antibodies for use in streptavidin-biotin based assay.
    Investigation of cell viability upon treatment with the naked antibody (black), antibody-Streptavidin-Biotin-Saporin conjugate (orange) or Saporin alone (grey). The ribosome inhibitor Saporin, unable to enter the cell alone, can be used to investigate antibody internalization by measuring viability (MTS) of antibody-SB-Saporin conjugate-treated cells.

  • Conjugation - Streptavidin Conjugation Kit&nbsp;- Lightning-Link&reg; (ab102921)
    Conjugation - Streptavidin Conjugation Kit - Lightning-Link® (ab102921)Image from Wang et al., Redox Biol. 24:101153; doi: 10.1016/j.redox.2019.101153. Reproduced under the Creative Commons license https://creativecommons.org/licenses/by/4.0/

    Wang, Xinzhi et al used Streptavidin Conjugation Kit - Lightning-Link® (ab102921) as part of examining. They used the kit to conjugate Streptavidin to recombinant proteins for use in bio-layer interferometry.
    Comparison of the binding activity between PepE and TrxR1 protein (red line), PepE and TrxR1 nSec498 protein (blue line), and PepA and TrxR1 protein (black line) through BLI analysis.

  • Streptavidin Conjugation Kit
    Streptavidin Conjugation Kit

    This illustration demonstrates a general procedure and will slightly vary dependent on the conjugate used.

プロトコール

  • Protocol Booklet - Chinese Version
  • Protocol Booklet

Click here to view the general protocols

データシートおよび資料

  • SDS download

  • Datasheet download

    Download

参考文献 (33)

ab102921 を使用した論文を発表された方は、こちらまでお知らせください。データシートに掲載させていただきます。

ab102921 は 33 報の論文で使用されています。

  • Pereira PM  et al. Super-beacons: Open-source probes with spontaneous tuneable blinking compatible with live-cell super-resolution microscopy. Traffic 21:375-385 (2020). PubMed: 32170988
  • Hoffmann RM  et al. Rapid conjugation of antibodies to toxins to select candidates for the development of anticancer Antibody-Drug Conjugates (ADCs). Sci Rep 10:8869 (2020). PubMed: 32483228
  • Sillerud LO  et al. Longitudinal monitoring of microglial/macrophage activation in ischemic rat brain using Iba-1-specific nanoparticle-enhanced magnetic resonance imaging. J Cereb Blood Flow Metab 40:S117-S133 (2020). PubMed: 32960690
  • Wang X  et al. Peperomin E and its orally bioavailable analog induce oxidative stress-mediated apoptosis of acute myeloid leukemia progenitor cells by targeting thioredoxin reductase. Redox Biol 24:101153 (2019). PubMed: 30909158
  • Wang Z  et al. Endothelial capture using antibodies and nanoparticles in human tissues: Antigen identification and liver segment imaging. Acta Biomater 97:474-489 (2019). PubMed: 31398471
View all Publications for this product

レビューと Q&A

Show All レビュー Q&A
レビューを送る 質問を送る

1-2 of 2 Abreviews or Q&A

Question

I came across your Easylink Biotin conjugation kit online and I would like to know if you have a way of determining your MSR after biotinylation. I would like to know the ratio of biotin per antibodies. Is this kit able to reach biotinylation levels between 6-8 biotins per antibodies?
I also would like to know if your Strepavidin Conjugation kit to conjugate antibodies provides a way to determine the ratios of strepavidin per antibodies. Is it a robust process that would conjugate all antibodies (polyclonals and monoclonals) in a consistent way?

Read More

Abcam community

Verified customer

Asked on Nov 16 2012

Answer

Thank you for contacting us.

With regard to the number of biotins, each antibody will be conjugated with multiple biotins, getting up to 6 or 8 biotins per antibody is feasible. It is important to remember that you only need one biotin attached to enable the biotinylated antibody to bind to streptavidin. There are assays available to measure biotinylation. However, in this case they will not be suitable as there is no separation step post conjugation.

With regard to Streptavidin, you can only confirm the streptavidin is attached to the antibody by size. The easiest way is to run an ID gel and measure the molecular weight. An antibody is 160KD and streptavidin is 53KD. A conjugate will therefore by 160 + 53KD (one strep) or 160 + 106 (two streps) or 160 + 159KD (3 streps). In reality you will get a mixture of all three. Also you must run a non-reducing gel otherwise you will get a mess due to all the possible combinations as both proteins are composed of subunits.

You can control the molar ratio by adding different amounts of antibody. This will guide the actual labelling to a desired ratio. For example, adding the following amounts of antibody will give different molar ratios:

100ug of antibody to 100ug of strep = 3 streps on each antibody

150ug of antibody to 100ug of strep = 2 streps on each antibody

300ug of antibody to 100ug of strep = 1 strep on each antibody

This is only a guide and it will not be absolute, as no reaction is 100% efficient.

As for the question “Is it a robust process that would conjugate all antibodies (polyclonals and monoclonals) in a consistent way?” If the same antibody, either monoclonal or polyclonal, is used in the same conditions (buffers, incubations times, etc) the results will be very reproducible. If labeling is carried out with different antibodies then there might some variation between results.

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 Nov 16 2012

Question

Hello I bought from you Streptavidin Conjugation kit – ab102921 – and I used 200ul of Ab (conc 500ug/ml) – so I had 100ug in 200ul and I realized that conjugation efficiency was not good (a lot of Ab`s were uncojugated). In recommended amount and volume of Ab for optimal results you suggest to have 100-200ug of Ab in max 100ul. So my question is – what should I do to obtain good efficiency with 200ul of Ab in concentration 500ug/ml Best wishes

Read More

Abcam community

Verified customer

Asked on Jan 12 2012

Answer

Thank you for your inquiry. I am sorry to hear that ab102921 did not deliver optimal results for you. I recommend to use the kit as instructed with a maximum of 100ul and 100ug of antibody. These kits are optimized for this specific protocol. There is the option of concentrating the antibody in order to be able to use 100ug for the reaction while not exceeding the maximum volume. I can recommend ab102778 for this. Click here (or use the following: https://www.abcam.com/Antibody-Concentration-And-Clean-Up-Kit-ab102778.html). The Easy Link Kits work by targeting 'available' amines on the antibody. They will also label proteins etc that have amines. This is the reason not to use more volume as stated and to use purified antibody. Otherwise the streptavidin will label the proteins in the buffer instead of the antibodies. Many antibodies come in buffers that contain additional protein to keep the antibody stable. I suggest to purify the antibody in this case. I can recommend ab102784 for purification. Click here (or use the following: https://www.abcam.com/Antibody-Purification-Kit-Protein-A-ab102784.html). I hope this information is helpful. Please do not hesitate to contact me again with any further questions.

Read More

Abcam Scientific Support

Answered on Jan 12 2012

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