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Lysosomal Intracellular Activity Assay Kit (ab234622) provides a proprietary Lysosome-Specific Self-Quenched Substrate which has low background fluorescence, high signal to background ratio and is pH insensitive.

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Images

Functional Studies - Lysosomal Intracellular Activity Assay Kit (AB234622), expandable thumbnail
  • Functional Studies - Lysosomal Intracellular Activity Assay Kit (AB234622), expandable thumbnail
  • Functional Studies - Lysosomal Intracellular Activity Assay Kit (AB234622), expandable thumbnail

Publications

Key facts

Detection method

Fluorescent

Sample types

Suspension cells, Adherent cells

Assay type

Cell-based

Reactive species

Mammals

What's included?

50 Test
Components
100X Cytochalasin D
1 x 50 µL
Buffer I
1 x 1.8 mL
Self-Quenched Substrate
1 x 1 Vial

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Lysosomal Intracellular Activity Assay Kit (ab234622) provides a proprietary Lysosome-Specific Self-Quenched Substrate which has low background fluorescence, high signal to background ratio and is pH insensitive.

Key facts

Detection method

Fluorescent

Sample types

Suspension cells, Adherent cells

Assay type

Cell-based

Reactive species

Mammals

Assay Platform

Flow cytometer, Fluorescence microscope

Storage

Shipped at conditions

Blue Ice

Appropriate short-term storage conditions

-20°C

Appropriate long-term storage conditions

-20°C

Storage information

-20°C

Notes

Lysosomal Intracellular Activity Assay Kit (ab234622) provides a proprietary Lysosome-Specific Self-Quenched Substrate which has low background fluorescence, high signal to background ratio and is pH insensitive. The substrate, acting as endocytic cargo, can be taken up by cells and degraded within an endo-lysosomal vesicle. The fluorescent signal is recovered from the Self-Quenched Substrate. The fluorescence signal, generated by degradation, is proportional to the intracellular lysosomal activity and can be measured using a fluorescence microscopy and/or flow cytometry. The kit includes Cytochalasin D, a cell-permeable inhibitor of the lysosomal membrane V-ATPase proton pump, that serves as an anti-lysosomal experimental control. This easy-to-use, non-radioactive kit allows imaging and accurate measurement of de-quenching substrate in cultured cells.

This product is manufactured by BioVision, an Abcam company and was previously called K448 Lysosomal Intracellular Activity Assay Kit. K448-50 is the same size as the 50 test size of ab234622.

Lysosomes are membrane-bound organelles important for various cellular processes. They contain hydrolytic enzymes that are utilized in the metabolism of some biomolecules. The extracellular cargo (e.g. nutrients toxins) binds to the cell membrane and is subsequently transported into membrane-bound endosomes for further degradation by lysosomes while intracellular components are transported to lysosomes through autophagy. Lysosomal dysfunction is associated with many human conditions such as aging and neurodegenerative disease.

Product promise

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Full details and terms and conditions can be found here:
Terms & Conditions.

3 product images

  • Functional Studies - Lysosomal Intracellular Activity Assay Kit (ab234622), expandable thumbnail

    Functional Studies - Lysosomal Intracellular Activity Assay Kit (ab234622)

    Release of self-Quenched Substrate in U937 cells.

    1x106 U937 cells were pretreated with vehicle or 1X Cytochalasin D for 1 hour. After pre-treatment, cells were incubated with Self-Quenched Substrate and the same concentration of Cytochalasin D for additional hour in medium supplemented with 0.5% FBS according to kit's protocol. Lysosomal staining with Lysosomal Associated Membrane Protein 1 (Lamp-1 RFP, a lysosomal marker). Cells were stained with nuclear dye for 10 minutes, washed with 1X PBS and mounted on the slide. Images were taken using a fluorescence microscope with a 60X objective lens.

  • Functional Studies - Lysosomal Intracellular Activity Assay Kit (ab234622), expandable thumbnail

    Functional Studies - Lysosomal Intracellular Activity Assay Kit (ab234622)

    Release of self-Quenched Substrate in U937 cells. 1x106 U937 cells were pretreated with or without 1X Cytochalasin D for 1 hr. After pretreatment, remove the medium, cells were incubated with Self-Quenced Substrate, and the same concentration of Cytochalasin D for another 1 hr in medium with 0.5% FBS according to kit's protocol. (A) Comparison of histograms from flow analysis showing the inhibition of De-Quenching of Substrate by Cytochalasin D in U937 cells (Black: negative control cells; Pink: in the presence of 1X Cytochalasin D; Green: without 1X Cytochalasin D).

  • Functional Studies - Lysosomal Intracellular Activity Assay Kit (ab234622), expandable thumbnail

    Functional Studies - Lysosomal Intracellular Activity Assay Kit (ab234622)

    Release of self-Quenched Substrate in U937 cells.

    1x106 U937 cells were pretreated with vehicle or 1X Cytochalasin D for 1 hour. After pre-treatment, cells were incubated with Self-Quenched Substrate and the same concentration of Cytochalasin D for additional hour in medium supplemented with 0.5% FBS according to kit's protocol. Images of U937 cells obtained using fluorescence microscope. Top: positive control cells treated Self-quenched substrate only. Bottom: negative control cells treated with 1X Cytochalasin D. U937 cells showing the release of Self-quenched substrate in the endocytotic vesicle (punctured pattern).

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Product protocols

For this product, it's our understanding that no specific protocols are required. You can:

Please note: All products are 'FOR RESEARCH USE ONLY. NOT FOR USE IN DIAGNOSTIC OR THERAPEUTIC PROCEDURES'.

For licensing inquiries, please contact partnerships@abcam.com