At the peak stage (18–20 days) of neurological impairment, NADPH oxidase activity was measured using a colorimetric NADP/NADPH assay kit (Abcam, Cambridge, UK) according to the manufacturer's instructions. Lumber spinal cords (n = 3 per group) were lysed in an assay buffer provided in the kit. The lysates were deproteinized by passing through a 10 kD Spin column (Abcam). The assay was performed in a 96-well plate and absorbance was measured with a microplate luminometer (Molecular devices, VERSAmax™ Tunable Microplate Reader San Jose, CA, USA) at 450nm. For every sample, NADPH oxidase activity was calculated as the difference between activities obtained in the presence and absence of NADPH.
Colorimetric nadp nadph assay kit
Manufactured by Abcam
Sourced in United States
The Colorimetric NADP/NADPH Assay Kit is a laboratory tool used to quantify the levels of NADP (nicotinamide adenine dinucleotide phosphate) and NADPH (the reduced form of NADP) in biological samples. It provides a colorimetric detection method for measuring these cofactors.
Automatically generated - may contain errors
Lab products found in correlation
7 protocols using colorimetric nadp nadph assay kit
1
Measuring NADPH Oxidase Activity
2
Colorimetric NADP/NADPH Assay
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
NADP/NADPH levels were assessed using a colorimetric NADP/NADPH assay kit (Abcam, Cambridge, MA, USA) following the manufacturer’s instructions. Cells were lysed in an assay buffer provided in the kit. The lysates were deproteinized by passing through a 10 kD Spin column (Biovision, Milpitas, CA, USA). The assay was performed in a 24-well plate, and absorbance was measured with a Multimode Plate Reader Victor X3, P (Perkin Elmer, Hopkinton, MA, USA) at 450 nm.
3
NADP/NADPH Colorimetric Assay
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
The NADP/NADPH levels were assessed using a colorimetric NADP/NADPH assay kit (Abcam, Cambridge, MA, USA), following the manufacturer’s instructions. The cells were lysed in the assay buffer provided in the kit. The lysates were deproteinized by passing through a 10-kD spin column (Biovision, Milpitas, CA, USA). Thereafter, the assay was performed in a 24-well plate, and the absorbance of the samples was measured using a Multimode Plate Reader Victor X3, P (Perkin Elmer, Hopkinton, MA, USA) at 450 nm.
4
Colorimetric NADP/NADPH Quantification
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
NADP/NADPH levels were assessed using a colorimetric NADP/NADPH assay kit (Abcam) following the manufacturer's instructions. Absorbance was measured with a Multimode Plate Reader Victor X3, P (PerkinElmer, Hopkinton, MA, USA) at 450 nm.
5
Quantifying Cellular NADP/H Levels
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Total nictoinamide adenine dinucleotide phosphate (NADP/H) and NADPH were measured using the NADP/NADPH colorimetric assay kit (Abcam). Equal number of sorted LSK cells (Live, lineage marker negative, cKit+Sca1+) were washed 3 times in ice-cold PBS and lysed in NADP/NADPH extraction buffer by performing 2 freeze/thaw cycles (20 minutes on dry ice followed by 10 minutes at room temperature). Lysates were centrifuged at 13,000g for 10 minutes at 4°C and the supernatant was retained. Lysate supernatant was split in half, with one half remaining on ice and the other half incubated at 60°C for 30 minutes to remove NADP+. Total NADP/H (NADPt) and NADPH only lysates were run in 96-well plates with freshly made standards as per the manufacturers’ instructions. NADP/NADPH ratio was calculated as (NADPt-NADPH)/NADPH.
6
NADP/NADPH Quantification in AML Cell Lines
7
NADP/NADPH Quantification in AML Cell Lines
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Total NADP/H and NADPH were measured using the NADP/NADPH colorimetric assay kit (Abcam). AML cell lines were seeded at 2x105 cells/ml one day prior to treatment with the indicated drugs (day 0). The following day (day 1), drugs were added to culture wells at the indicated concentration and cells were harvested after 8 hours. All cells were collected from the wells and washed three times in ice-cold PBS. Cells were lysed in NADP/NADPH extraction buffer by performing two freeze/thaw cycles (20 mins on dry ice followed by 10 mins at room temperature). Lysates were centrifuged at 13,000g for 10minutes and the supernatant was retained. Lysate supernatant was split in half, with one half remaining on ice and the other half incubated at 60C for 30mins to remove NADP+. Total NADP/H (NADPt) and NADPH only lysates were run in 96 well plates with freshly made standards as per the manufacturers’ instructions. NADP/NADPH ratio was calculated as (NADPt-NADPH)/NADPH.
About PubCompare
Our mission is to provide scientists with the largest repository of trustworthy protocols and intelligent analytical tools, thereby offering them extensive information to design robust protocols aimed at minimizing the risk of failures.
We believe that the most crucial aspect is to grant scientists access to a wide range of reliable sources and new useful tools that surpass human capabilities.
However, we trust in allowing scientists to determine how to construct their own protocols based on this information, as they are the experts in their field.
Ready to get started?
Sign up for free.
Registration takes 20 seconds.
Available from any computer
No download required
Revolutionizing how scientists
search and build protocols!