Key picks and particulars

  • Sensitivity: 40 cells/effectively
  • Sample choice: Adherent cells, Suspension cells
  • Detection technique: Colorimetric
  • Assay choice: Sandwich (qualitative)

5-bromo-2-deoxyuridine (BrdU) is a pyrimidine analog. It’s going to get built-in into the newly synthesized DNA of proliferating cells as a substitute of thymidine. Biovision’s BrdU Cell Proliferation Assay Bundle deal detects built-in BrdU using a mouse anti-BrdU antibody. An anti-mouse HRP-linked secondary antibody is used to detect the anti-BrdU antibody sure to BrdU, which is adopted by addition of TMB (a HRP substrate). The extent of coloration enchancment is proportional to the quantity of BrdU built-in into the cells and will presumably be utilized straight as an indicator of cell proliferation. In distinction with totally completely completely different cell proliferation assays, this bundle deal detects solely the proliferating cells and by no means the seeded cells. This terribly delicate, non-radioactive bundle deal detects as a lot so much a lot much less as 50-100 proliferating cells.

Background

Halogenated nucleotides such due to the pyrimidine analog bromodeoxyuridine (BrdU) are useful for labeling nascent DNA in residing cells and tissues. BrdU turns into built-in into replicating DNA as a substitute of thymidine and subsequent immunodetection of BrdU using specific monoclonal antibodies permits labeling of cells in S a part of the cell cycle. After pulse-labeling cells or tissues with bromodeoxyuridine, BrdU (Bu20a) Mouse mAb may presumably be utilized to detect BrdU built-in into single stranded DNA.

Product Description

The BrdU Cell Proliferation Assay Bundle deal detects 5-bromo-2’-deoxyuridine (BrdU) built-in into cellular DNA all by means of cell proliferation using an anti-BrdU antibody. When cells are cultured with labeling medium that comes with BrdU, this pyrimidine analog is built-in as a substitute of thymidine into the newly synthesized DNA of proliferating cells.

Brdu Cell

Brdu Cell

After eradicating labeling medium, cells are fixed and the DNA is denatured with our fixing/denaturing reply. Then a BrdU mouse mAb is added to detect the built-in BrdU (The denaturing of DNA is essential to bolster the accessibility of the built-in BrdU to the detection antibody). Anti-mouse IgG, HRP-linked antibody is then used to acknowledge the sure detection antibody. HRP substrate TMB is added to develop coloration. The magnitude of the absorbance for the developed coloration is proportional to the quantity of BrdU built-in into cells, which is a direct indication of cell proliferation.

Product Consists of Quantity (with Rely) Reply Shade
BrdU 1 x 150 µl
Fixing/denaturing Reply 2 x 25 ml
BrdU Mouse Detection mAb 1 x 500 µl Inexperienced
Anti-mouse IgG, HRP-Linked Antibody 1 x 500 µl Pink
Detection Antibody Diluent 1 x 50 ml Inexperienced
HRP Diluent 1 x 50 ml Pink
20X Wash Buffer 1 x 50 ml
TMB Substrate 7004 1 x 50 ml
STOP Reply 7002 2 x 25 ml

 

BrdU Cell Proliferation ELISA Kit (colorimetric)

BrdU Cell Proliferation ELISA Package (colorimetric)

METABOLIC PROLIFERATION ASSAYS

Assays that measure metabolic put together are acceptable for analyzing proliferation, viability, and cytotoxicity. The low value of tetrazolium salts an identical to MTT, XTT, and WST-1 to colored formazan compounds or the bioreduction of resazurin occurs solely in metabolically energetic cells. Actively proliferating cells improve their metabolic put together, whereas cells uncovered to toxins might have decreased put together.

MTT Cell Proliferation Assays

MTT (3-[4,5-dimethylthiazol-2-yl]-2,5- diphenyltetrazolium bromide; thiazolyl blue) is a water soluble tetrazolium salt yielding a yellowish reply when prepared in media or salt picks lacking phenol pink. Dissolved MTT is remodeled to an insoluble purple formazan by cleavage of the tetrazolium ring by dehydrogenase enzymes. This water-insoluble formazan is probably solubilized using isopropanol or totally completely completely different solvents, and the dissolved gives is measured spectrophotometrically using absorbance as a function of focus of remodeled dye.

XTT Cell Proliferation Assays

In distinction to MTT, the cleavage product of XTT is soluble in water; a solubilization step is subsequently not required. The tetrazolium salt XTT is cleaved to formazan by a elaborate cellular mechanism. This bioreduction occurs in viable cells solely, and is expounded to NAD(P)H manufacturing by way of glycolysis. The amount of formazan dye frequent straight correlates to the number of metabolically energetic cells contained within the personalized.

WST-1 Cell Proliferation Assays

The frequent tetrazolium salt WST-1 is cleaved to a soluble formazan by a elaborate cellular mechanism that occurs utterly on the cell flooring. This bioreduction is particularly relying on the glycolytic manufacturing of NAD(P)H in viable cells. The amount of formazan dye frequent straight correlates to the number of metabolically energetic cells contained within the personalized.

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