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Gold Conjugation Kit (40nm, 20 OD) (ab154873)

Submit a review Q&A (3)References (12)
Conjugation - Gold Conjugation Kit (40nm, 20 OD) (ab154873)
  • Lateral flow assay - Gold Conjugation Kit (40nm, 20 OD) (ab154873)
  • Lateral flow assay - Gold Conjugation Kit (40nm, 20 OD) (ab154873)
  • Lateral flow data – Gold Conjugation Kit vs traditional gold nanoparticle passive absorption techniques
  • GOLD Conjugation Kit (40nm, 20 OD)

Overview

  • Product name

    Gold Conjugation Kit (40nm, 20 OD)
    See all Gold kits

  • Product overview

    Abcam’s Gold Conjugation Kit allows antibodies or proteins to be covalently attached to ultra-stable Gold nanoparticles at very high OD quickly and easily. The nanoparticles in the Abcam’s Gold Conjugation Kit have a protective surface coat that can withstand the most extreme conditions (e.g. 2.5M NaOH at 70°C for > 1 hour). The hands-on time for the Abcam Gold conjugation procedure is around 2 minutes and the conjugate is ready to use within 20 minutes. The researcher simply pipettes the biomolecule into a vial containing the nanoparticles of the Gold Conjugation Kit.


    The nanoparticles in this kit are supplied as a freeze-dried mixture. The conjugation reaction is initiated simply by reconstituting the freeze-dried nanoparticles with the antibody, which becomes attached (through lysine residues) to the surface of the nanoparticles.


    The resulting covalent conjugates are more stable than those prepared by passive adsorption methods. Moreover, unlike passive methods, the coating process is independent of the isoelectric point of the antibody, avoiding the need for extensive trials at different pH values. All antibodies can be labelled at a single pH.


    Learn more about buffer compatibility, protein/secondary antibody conjugation and labeling chemistry in our FAQs.


    Benefits


    Easy and rapid conjugation – only 2 minutes hands-on time and 100% recovery of materials


    Site-specific labelling – Antigen binding sites of antibodies are free to bind the target molecule


    Proprietary surface coating prevents metal-protein interactions, and enables covalent attachment to the Gold – Stable conjugates formed


    Fully scalable – Easy transfer from R&D to manufacturing


    Uniform spherical shape and narrow size distribution – Consistent high quality and excellent batch-to-batch reproducibility


    Buffer requirements:


    The biomolecule to be conjugated should ideally be in 10 mM amine-free buffer (e.g. MES, MOPS, HEPES), pH range 6.5 to 8.5. Sugars have no effect on conjugation efficiency. For incompatible buffers and low antibody concentrations, use our rapid antibody purification and concentration kits for Nanoparticles. To learn more about incompatible buffers, please refer to the protocol booklet.

  • Notes

    This product is manufactured by Expedeon, an Abcam company, and was previously called 40nm InnovaCoat® GOLD. 230-0010 is the same as the 10 x 1 µg size. 230-0015 is the same as the 1 x 10 µg size. 230-0005 is the same as the 3 x 1 µg size.

    The 3 and 10 Test Conjugation Kits are designed to label 12 µl of antibody per vial.

    The 1 Test Conjugation Kit is designed to label 120 µl of antibody per vial.

Properties

  • Storage instructions

    Store at -20°C. Please refer to protocols.
  • Components 3 x 1 µg 10 x 1 µg 1 x 10 µg
    ab273949 - Gold 40nm 3 x 1µg 10 x 1µg 1 x 10µg
    ab273943 - Gold Antibody Diluent 1 x 1ml 1 x 4ml 1 x 4ml
    ab273942 - Gold Quencher Reagent 1 x 700µl 1 x 700µl 1 x 700µl
    ab273941 - Gold Reaction Buffer 1 x 750µl 1 x 750µl 1 x 750µl

  • Research areas

Images

  • Conjugation - Gold Conjugation Kit (40nm, 20 OD) (ab154873)
    Conjugation - Gold Conjugation Kit (40nm, 20 OD) (ab154873)Image from Guzman et al., Biosensors (Basel), 10(10):130; doi: 10.3390/bios10100130.Reproduced under the Creative Commons license https://creativecommons.org/licenses/by/4.0/

    Guzman, John Mello Camille C., Sheng-Min Hsu, and Han-Sheng Chuang used Gold Conjugation Kit (40nm, 20 OD) (ab154873) as part of developping a colorimetric diagnostic capillary device for the rapid detection of biomarkers. They used the kit to conjugate Gold to probe rabbit polyclonal anti-LCN antibody for use in the device.
    (a) Illustrations for preparing sandwiched immunocomplex and detection process. (b) Sandwiched immunocomplex detection process. (c-g) Quantitative assessment of the detection platform: (c) grayscale images of diagnostic tool inside the detection box; (d) calibration of the grayscale intensity of sandwiched immunocomplex with respect to different lipocalin-1 (LCN-1) concentrations ranging from 100 pg/mL to 1 µg/mL; (e) binding specificity of the capture and probe particles in the presence of different antigens; (f) binding stability of sandwiched immunocomplex with respect to different times; (g) reliability of the detection platform. Comparison of LCN-1 concentration detection results between the proposed diagnostic tool and spectrophotometer. The symbols "*", "**", "***", "ns" denote p < 0.05, p < 0.01, p < 0.001, and p > 0.05, respectively, under student's t test (n = 5). The error bars represent standard deviation.

  • Lateral flow assay - Gold Conjugation Kit (40nm, 20 OD) (ab154873)
    Lateral flow assay - Gold Conjugation Kit (40nm, 20 OD) (ab154873)Image from Lee et al., Scientific reports 6 (2016): 28237. Sci Rep., 6:28237; doi: 10.1038/srep28237. Reproduced under the Creative Commons license https://creativecommons.org/licenses/by/4.0/

    Lee, Seoho, et al used Gold Conjugation Kit (40nm, 20 OD) (ab154873) as part of examining low-cost point-of-care quantification of vitamin B12 concentrations. They used the kit to conjugate Gold to anti-vitamin B12 IgG for use in competitive lateral flow assay.
    (A) Colorimetric variation of the test and control line regions on the silver enhanced B12 lateral flow test strip at different known concentrations of standard vitamin B12 samples (B) image processing algorithm used by the NutriPhone platform (C) Test and control line signals detected by the NutriPhone app as local intensity minima for 0 and 369 pmol/L of B12 standard samples (D) Calibration curve showing the T/C ratios of the colorimetric signals at different standard B12 concentrations.

  • Lateral flow assay - Gold Conjugation Kit (40nm, 20 OD) (ab154873)
    Lateral flow assay - Gold Conjugation Kit (40nm, 20 OD) (ab154873)Image from Vemulapati et al., Sci Rep., 7(1):14142; doi: 10.1038/s41598-017-13044-5. Reproduced under the Creative Commons license https://creativecommons.org/licenses/by/4.0/
    Vemulapati, S., et al used Gold Conjugation Kit (40nm, 20 OD) (ab154873) as part of examining vitamin D3 Deficiency. They used the kit to conjugate Gold to monoclonal anti-25(OH)D3 IgG antibody for use in lateral flow assay.
    Vitamin D3 Lateral Flow Assay. (A) Image and schematic of the 25(OH)D3 strip architecture and components. (B) Image and intensity plot of a participant with low Vitamin D3 and high T/C ratio (C) Image and intensity plot of a participant with healthy levels of 25(OH)D3 and low T/C ratio.
  • Lateral flow data – Gold Conjugation Kit vs traditional gold nanoparticle passive absorption techniques
    Lateral flow data – Gold Conjugation Kit vs traditional gold nanoparticle passive absorption techniques

    Antibody conjugation using the Gold Conjugation Kit vs traditional gold nanoparticle passive absorption techniques with uncoated gold nanoparticles, showing both enhanced signal intensity and improved specificity. 40 nm Gold particles were labeled with anti-IgA antibody and used to measure IgA concentration in a lateral flow inhibition assay, with IgA bound to a lateral flow strip.

  • GOLD Conjugation Kit (40nm, 20 OD)
    GOLD Conjugation Kit (40nm, 20 OD)

    Please see the protocol booklet for a detailed method. 

References (12)

Publishing research using ab154873? Please let us know so that we can cite the reference in this datasheet.

ab154873 has been referenced in 12 publications.

  • Lee JH  et al. A novel rapid detection for SARS-CoV-2 spike 1 antigens using human angiotensin converting enzyme 2 (ACE2). Biosens Bioelectron 171:112715 (2021). PubMed: 33099241
  • Elter A  et al. Carbohydrate binding module-fused antibodies improve the performance of cellulose-based lateral flow immunoassays. Sci Rep 11:7880 (2021). PubMed: 33846482
  • Guzman JMCC  et al. Colorimetric Diagnostic Capillary Enabled by Size Sieving in a Porous Hydrogel. Biosensors (Basel) 10:N/A (2020). PubMed: 32977557
  • Srinivasan B  et al. Rapid diagnostics for point-of-care quantification of soluble transferrin receptor. EBioMedicine 42:504-510 (2019). PubMed: 30885726
  • Chuang HS  et al. Enhanced diffusometric immunosensing with grafted gold nanoparticles for detection of diabetic retinopathy biomarker tumor necrosis factor-a. Biosens Bioelectron 101:75-83 (2018). PubMed: 29040917
View all Publications for this product

Customer reviews and Q&As

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1-3 of 3 Abreviews or Q&A

Question



Inquiry: Respected Sir/Madam The Gold conjugation kits are available at various sizes of the gold nanoparticles. What will be the effect of these sizes? What will be the difference in the signals? Kindly help me choose the most suitable size for a lateral flow chromatography colorimetric detection.

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Answer

Thank you for getting in touch.

I can confirm that 40nm gold nanoparticles are the most popular choice for lateral flow assays. This is due to an optimal combination of high contrast colour which appears cherry red and surface area for effective and efficient analyte testing,

30nm and 60nm gold nanoparticle also have specific advantages depending on the design and application of the lateral flow test.

30nm gold nanoparticles have a darker red appearance that improves contrast against white lateral flow assay membranes compared to 40 nm spheres. With increased contrast, due to the smaller size 30 nm gold nanoparticle require more conjugated antibodies to achieve an equivalent mass concentration.

60nm gold nanoparticles have a lighter red appearance which trades contrast efficiency for increased optical absorbance per particle. Thus, less particles are needed to produce a measurable reading. This means 60 nm gold are ideal for immunoassays with low target analyte concentration samples, or when the targeting moiety is very expensive.


In addition to this, the following information may be helpful:

In principle, the smaller gold particles produce a higher labelling intensity. This is due to reduced steric hindrance to antigen detection.  For example, a 1nm gold particle, attached to the Fc region should not significantly affect antibody binding. A 20nm particle, however, while being more visible, will produce a greater steric hindrance. In addition, the increased charge repulsion between larger particles reduces the number of labelled antibodies gaining access to the target antigen.The surface of gold nanoparticles have free electron which resonate with the oscillating electric field of a light ray propagating near it, creating a concerted oscillation of electron charge called surface plasmons.

These surface plasmons will cause absorption and reflection of lights, and these absorption and reflection are dependent on the size of the gold nanoparticle. For smaller gold nanoparticle, the reflected light is red – whereas the bigger nanoparticle will start to absorb redder wavelength and reflects more blue wavelength, yielding purple or more blue colour. Since larger nanoparticles have a higher magnitude of absorption and available surface area for antibody conjugation, they can provide better assay sensitivity. However, when larger nanoparticles are used, absorption in the longer wavelengths reduces the contrast on the test strip.


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Question

Tengo una duda sobre el kit, no entiendo a qué se refiere el término "OD", significa densidad óptica? en el caso de que sea densidad óptica, a qué longuitud de onda hbría que medirlo? cuál sería la equivalencia con su concentración?

Read More
Answer



El termino OD se refiere efectivamente a la densidad óptica para el oro, que es la manera más común de expresar la concentración de las nano partículas de oro. La longitud de onda se mide a 530nm.

Me temo que no podemos proporcionar el equivalente en concentración, pero quizás os ayude saber que hay aproximadamente 9x1010 nano partículas por 50µl de conjugado.

Read More

Question

Dear Tech Support Team,

Our customer is interested in ab154873 but he is concerned about high isoelectric point of his protein (PI=9). Please advise if ab154873 is suitable for the conjugation for this kind of the protein (it is a core of Hepatitis C).

Thanks in advance for your assistance and reply.

Read More
Answer

Thank you for your enquiry regarding ab154873.
I can confirm that the high isoelectric point of the protein should not be a problem when using ab154873: EasyLink GOLD Conjugation Kit.
The main points to bear in mind about the protein buffer prior to labeling with this kit are:
● Protein must be purified
● Avoid amino acids (e.g. glycine)
● Avoid other primary amines (e.g. Tris)
● Avoid thiols (e.g. mercaptoethanol, DTT)
● Avoid carboxylic acids (e.g. EDTA)

Your customer should also note that the protocol will be the same as when labeling an antibody. However, if the protein is significantly larger or smaller than a typical IgG (about 160kD) they might want to explore varying ratios of protein.
The optimum amount of antibody or protein in this case (which will influence the number of antibody/protein molecules per particle) may be application-dependent and you may need to conjugate different amounts of antibody to optimize your assay.
I would advise your customer to download the protocol booklet (click on the blue hyperlink bottom section on the on-line datasheet) and read the document carefully.
The initial amount of antibody recommended corresponds to 10ug antibody per ml of 10 OD gold, which is about half of that normally used for passive (non-covalent) conjugations. However lower or higher concentrations can be explored as there is no risk of aggregation because of the protective surface coat.
I hope that this helps. If you need any further assistance in the future, please do not hesitate to contact me.

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