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    products/conjugation-kits/fitc-conjugation-kit-lightning-link-ab102884.pdf

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FITC Conjugation Kit - Lightning-Link® (ab102884)

  • Datasheet
  • SDS
Reviews (3)Q&A (45)References (42)

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Immunohistochemistry (PFA perfusion fixed frozen sections) - FITC Conjugation Kit - Lightning-Link® (ab102884)
  • Immunocytochemistry - FITC Conjugation Kit- Lightning-Link
  • Flow Cytometry - FITC Conjugation Kit- Lightning-Link
  • Conjugation - FITC Conjugation Kit (ab102884)

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Overview

  • Product name

    FITC Conjugation Kit - Lightning-Link®
    See all FITC kits
  • Product overview

    FITC Conjugation Kit / FITC Labeling Kit ab102884 uses a simple and quick process for FITC labeling / conjugation of antibodies. It can also be used to conjugate other proteins or peptides. Learn about our antibody labeling kits and their advantages.


    We recommend FITC Conjugation Kit ab188285 as an alternative this kit using a faster, newer protocol.


    To conjugate an antibody to FITC using this kit:
    - add modifier to antibody and incubate for 3 hrs
    - add quencher and incubate for 30 mins
    The FITC 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 FITC.


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

  • Notes

    This product is manufactured by Expedeon, an Abcam company, and was previously called Lightning-Link® Fluorescein Labeling Kit. 707-0005 is the same as the 100 µg size. 707-0010 is the same as the 3 x 100 ug size. 707-0030 is the same as the 3 x 10 ug size. 707-0015 is the same as the 1 mg size.

    Amount and volume of antibody for conjugation to FITC 

     Kit size

    Recommended 
    amount of antibody1 

    Maximum 
    amount of antibody

    Maximum antibody 
    volume2

    3 x 10 µg

    3 x 10 µg

     3 x 20 µg

    3 x 10 µL

    100 µg

    1 x 100 µg

    1 x 200 µg

    1 x 100 µL

    3 x 100 µg

    3 x 100 µg 

    3 x 200 µg

    3 x 100 µL

    1 mg

    1 x 1 mg

    1 x 2 mg

    1 x 1 mL

     

    1 Using the maximum amount of antibody may result in less labelling per antibody.

    2 Ideal antibody concentration is 1mg/ml. 0.5 - 1 mg/ml can be used if the maximum antibody volume is not exceeded. Antibodies > 2 mg/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.

    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.

Properties

  • Storage instructions

    Store at -20°C. Please refer to protocols.
  • Components 1 mg 100 µg 3 x 10 µg 3 x 100 µg
    ab274132 - FITC mix 1 x 1mg 1 x 100µg 3 x 10µg 3 x 100µg
    ab274106 - Modifier reagent 1 x 200µl 1 x 200µl 1 x 200µl 1 x 200µl
    ab274133 - Quencher reagent 1 x 200µl 1 x 200µl 1 x 200µl 1 x 200µl
  • Research areas

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

Associated products

  • Related Products

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

Images

  • Immunohistochemistry (PFA perfusion fixed frozen sections) - FITC Conjugation Kit&nbsp;- Lightning-Link&reg; (ab102884)
    Immunohistochemistry (PFA perfusion fixed frozen sections) - FITC Conjugation Kit - Lightning-Link® (ab102884)Image from Wills, Jonathan, et al., PLoS One, 6(3): e17953; doi: 10.1371/journal.pone.0017953. Reproduced under the Creative Commons license https://creativecommons.org/licenses/by/4.0/

    Wills, Jonathan, et al used FITC Conjugation Kit - Lightning-Link® (ab102884) as part of examining the aggregation and co-localization of α-Syn with pGSK-3β and p-Tau in the striatum of A53T α-Syn mutant mice (A53T Tg) and wild-type mice (WT). They used the kit to conjugate FITC to Anti-p-GSK-3β (pY216) antibody for use in immunohistochemistry (PFA perfusion fixed frozen sections).
    Right panels constitute merged image of left panels. Sections of striatum of A53T Tg and age-matched WT mice were stained with anti-α-Syn antibody conjugated to Texas Red (red) and anti-p-GSK3β conjugated to FITC (green). Nuclei were stained with DAPI (blue).

  • Immunocytochemistry - FITC Conjugation Kit- Lightning-Link
    Immunocytochemistry - FITC Conjugation Kit- Lightning-LinkImage from Lim, Tony KY, et al., J Neurosci., 35(8):3346-59, doi: 10.1523/JNEUROSCI.4040-14.2015. Reproduced under the Creative Commons license https://creativecommons.org/licenses/by/4.0/

    Lim, Tony KY, et al used FITC Conjugation Kit - Lightning-Link® (ab102884) as part of examining traumatic nerve injury. They used the kit to conjugate FITC to mouse anti-neurofilament 160/200  for use in immunohistochemistry.B, Contralateral nerves were stained for hypoxyprobe-1, DAPI, and neurofilament 160/200 (NF-M/L, axons). E, Ipsilateral nerves were subjected to the same staining. The area in proximity to the injury site is shown. Hypoxia, as shown by hypoxyprobe-1 staining, is observed in a region close to the injury site, in which axons, macrophages, and Schwann cells are all found to be hypoxic. This hypoxic region, in which nerve fibers were ligated, is delineated by the white dotted line. Areas outside of the dotted regions containing nonligated nerve fibers still have more hypoxyprobe deposition than contralateral nerves.

  • Flow Cytometry - FITC Conjugation Kit- Lightning-Link
    Flow Cytometry - FITC Conjugation Kit- Lightning-LinkImage from Weagel, Evita G., et al., Cancer cell international 18.1 (2018): 1-14. Reproduced under the Creative Commons license https://creativecommons.org/licenses/by/4.0/

    Weagel, Evita G., et al used FITC Conjugation Kit - Lightning-Link® (ab102884) as part of examining thymidine kinase 1 in lung, breast, and colorectal malignancies. They used the kit to conjugate FITC to three custom and a commercial anti-human thymidine Kinase 1 (TK1) antibodies for use in flow cytometry.
    Membrane TK1 expression in of colon, breast, and lung cancer cell lines. Flow cytometry analysis of cell lines treated with anti-TK1 antibodies. a Quantification of TK1 expression on the cell membrane of HT-29 and SW620 cell lines stained with FITC or APC-conjugated anti-TK1 antibodies. b Quantification of TK1 expression on the cell membrane of MCF7 and MDA-MB-231 cell lines. The top bar graph shows MCF7 and MDA-MB-231 cell lines stained with FITC or APC-conjugated anti-TK1 antibodies. c Quantification of TK1 expression on the cell membrane of NCI-H460 and A549 cell lines stained with FITC or APC-conjugated anti-TK1 antibodies. Statistical analysis was performed by comparing the mouse isotype control fluorescent levels to those of A72, A74, CB1, or ab91651. *P ≤ 0.05; **P ≤ 0.005; ***P ≤ 0.001; ns = P > 0.05

  • Conjugation - FITC Conjugation Kit (ab102884)
    Conjugation - FITC Conjugation Kit (ab102884)

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

Protocols

  • Protocol Booklet - Chinese Version
  • Protocol Booklet

Click here to view the general protocols

Datasheets and documents

  • SDS download

  • Datasheet download

    Download

References (42)

Publishing research using ab102884? Please let us know so that we can cite the reference in this datasheet.

ab102884 has been referenced in 42 publications.

  • Huang KY  et al. Humanized COVID-19 decoy antibody effectively blocks viral entry and prevents SARS-CoV-2 infection. EMBO Mol Med 13:e12828 (2021). PubMed: 33159417
  • Micheva-Viteva SN  et al. Increased Mortality in Mice following Immunoprophylaxis Therapy with High Dosage of Nicotinamide in Burkholderia Persistent Infections. Infect Immun 87:N/A (2019). PubMed: 30323029
  • Liu G  et al. Fibulin-1c regulates transforming growth factor-ß activation in pulmonary tissue fibrosis. JCI Insight 5:N/A (2019). PubMed: 31343988
  • Dumigan A  et al. A Porcine Ex Vivo Lung Perfusion Model To Investigate Bacterial Pathogenesis. mBio 10:N/A (2019). PubMed: 31796543
  • Zhdanov DD  et al. Murine regulatory T cells induce death of effector T, B, and NK lymphocytes through a contact-independent mechanism involving telomerase suppression and telomere-associated senescence. Cell Immunol 331:146-160 (2018). PubMed: 29935763
View all Publications for this product

Customer reviews and Q&As

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1-10 of 48 Abreviews or Q&A

FITC Conjugation Kit

Excellent Excellent 5/5 (Ease of Use)
Abreviews
Abreviews
abreview image
We conjugated FITC to BSA protein. Then, the protein was bound to our microparticles for viewing. The left image is a brightfield image of the microparticles and the right image is the microparticles with BSA-FITC conjugate bound to them.
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 17 2020

FITC Conjugated full length EpCAM Protein

Excellent Excellent 5/5 (Ease of Use)
Abreviews
Abreviews
FITC was succesfully Conjugated with Human EpCAM Protein. It was quite useful.
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 02 2015

Better results than with secondary antibody.

Excellent Average 3/5 (Ease of Use)
Abreviews
Abreviews
The kit was easy enough for me to hand this off to an undergraduate, albeit with some explanation of the directions. These were a tad bit confusing at first, and not really clear on the volume of antibody to add (they give you a couple options since the Ab concentration of course matters, but didn't make it clear they were different options).
Otherwise the kit work fantastically with some store bought, pure, concentrated antibody! It actually gave better results than our previous method of using a conjugated secondary antibody when used in Flow Cytometry after doing an Intracellular Stain.
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.

Hans Wilms

Verified customer

Submitted Jun 03 2014

Question

Is there a difference in sensitivity between these 2 types of antibody labeling kits?

Read More

Abcam community

Verified customer

Asked on Sep 04 2014

Answer

There is no difference in sensitivity between the rapid (eg. ab188285) and the standard (eg. ab102884) labeling kits. The only difference between the two is the incubation time required for the reaction.

Read More

Jeremy Kasanov

Abcam Scientific Support

Answered on Sep 04 2014

Question





One of our customers is looking for an Anti-Enterococcus feacalis Ab, they would prefer that the Ab will be labeled by any florescence dye (FITS for example ).

Unfortunately I didn't find, is it correct or I missed it?

We offer them the Ab- Anti-Enterococcus antibody #ab19980 .On the detailed description it is written that the Immunogen is , E.faecalis,and on the alternative names are E. faecium antibody, E.faecalis antibody.

Base on this is there any information about cross-reactivity of the Ab between the different types?

Read More

Abcam community

Verified customer

Asked on Mar 19 2014

Answer


This antibody will detect the whole cells, and because the antiserum has not been absorbed (only Protein A purified) it may react with related microorganisms.


As this antibody is suitable for conjugation purpose according to the datasheet, I would like to sduggest your customer is having a look at our FITC conjugation kit.


Read More

Abcam Scientific Support

Answered on Mar 19 2014

Question

Inquiry: Dear Abcam, I have a novel protein that has been fused to Fc. I was wondering if your easylink antibody kits would be able to create a protein:fluorochrome conjugate if there is an N-terminal Fc fusion?

Read More

Abcam community

Verified customer

Asked on Aug 22 2013

Answer

Thank you for your enquiry.

The EasyLink conjugation kits are designed so that they can be used to label proteins other than antibodies. In this case you should be able to label the Fc fused protein in the same way as labeling an antibody. However as the size of their protein is likely different to that of an average antibody, please find advice we give in these situations below:

As the protocols provided were optimised for labeling IgGs, we would recommend to adjust the amount of material added to the label vial to allow for molecular weight difference. This should be done without changing the volume added to the vial, as this could affect the conjugation efficiency. As a rough guideline, we would recommend changing the amount of material proportionally to the size difference with IgGs. An average IgG is about 160kD, therefore for a target that is ½ the size of an antibody (about 80kD), add ½ as much to the vial.

Finally, please note the usual EasyLink requirements will still apply: the target should be purified, in a suitable amine free buffer and not contain any additives such as Azide, BSA, Tris or Glycine.

I hope that this helps. If you have any additional questions please do not hesitate to contact me.

Read More

Sam Washer

Abcam Scientific Support

Answered on Aug 22 2013

Question



Inquiry: In labeling an antibody with your Easylink, I would like to know to which residue you are attaching the reagent. How do you know it isn't attaching to the FAB portion?

Read More

Abcam community

Verified customer

Asked on Dec 19 2012

Answer

Thank you for contacting us.

The labeling kit works by targeting free amines groups on your antibody. We therefore cannot guarantee that the label will not bind to the FAB area, however this would be the case for any other amine orientated labeling technology. Although this might seem like an issue , it does not mean that the FAB will become unreactive, and we see successful conjugations in the great majority of cases.

The amine binding would only become an issue if there is an amine group within the antibody binding site itself.

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 19 2012

Question

Dear Representative,

I am interested in buying few products from your company which i will be ordering frequently if satisfied.
1. Anti-biotin Antibody (ab36406)
2. Anti-RAd54B (ab57036)
are these antibodies glycosylated?

3. easy link FITC conjugation kit (ab102884)
4. easy linkcy5 conjugation kit (ab102876)
i observed that no purification is required after conjugating the labels to antibodies. what is the basis of such reliability? Does all labels bind to the Abs and there is no free label?

Please answer my highlighted doubts and send me an electronic quote for the below mentioned address giving any discount (if applicable).

Read More

Abcam community

Verified customer

Asked on Nov 16 2012

Answer

Thank you for your inquiry and your interest in our products.

I will have to double-check with the lab regarding glycosylation of the antibodies since I do not have this information readily at hand. I will let you know once I have received this information.

For the EasyLink kits, there is a quenching step which removes any excess label and reduces background. You can find more information about the labeling process here: https://www.abcam.com/index.html?pageconfig=resource&rid=13148&source=pagetrap&viapagetrap=easylink

Additional FAQs regarding these kits can be found here: https://www.abcam.com/index.html?pageconfig=resource&rid=13156

I will email you a quote for the 4 products. Currently we are running a promotion where if you buy any primary, you can get a RabMAb for free! Just mention the code RABMAB-XBSMG on the order and the cost of the rabbit monoclonal antibody will be removed from the order.

I hope this information helps. I will be in touch again with more information regarding the antibodies you requested.

Read More

Abcam Scientific Support

Answered on Nov 16 2012

Question

How much FITC (moles, ideally) is in each vial of the 3 x 100-200ug conjugation kit? We hope to achieve a ratio of 2 FITC molecules perIgG molecule. Is there any data for a reference antibody, demonstrating the number of FITC molecules conjugated per IgG, for a given starting concentration of IgG (e.g., 1mg/ml)and a given formulation (e.g., PBS, azide-free, carrier--free)?

Read More

Abcam community

Verified customer

Asked on Sep 24 2012

Answer

Thank you for your interest in the FITC conjugation kit. Unfortunately, it will be difficult to achieve a consistent ratio of FITC : IgG without extensive testing of the kit to determine the appropriate conditions. We have not done this testing, as the kit was developed for ease of producing a conjugate that simply gives a bright signal, rather than producing a conjugate of a known ratio. Further, the components of the reagents are considered proprietary, which makes knowing where to start difficult. To do what you want, I think it will make more sense to work with a standard FITC conjugation protocol, using known reagents and quantities, and testing various combination of FITC and antibody.

Read More

Abcam Scientific Support

Answered on Sep 24 2012

Question

What is the CAS number of FITC molecules used in kit ab102884?

Read More

Abcam community

Verified customer

Asked on Sep 21 2012

Answer

Thank you for contacting us.

The fluorescein in ourKit is not simply fluorescein and it is not FITC either. Our fluorescein has been specially treated with proprietary Easy-Link chemistry to make the unique one-step conjugation possible.

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

Use our products? Submit an Abreview. Earn rewards!
https://www.abcam.com/abreviews

Read More

Abcam Scientific Support

Answered on Sep 21 2012

1-10 of 48 Abreviews or Q&A

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