I'm writing to you because I'm struggling with the WB to detect this protein. Even though it's in the right MW and not unspecific bands, I'm not able to obtain a band but a kind of spot with some smear that is difficult for the quantification. Please, could you send me a detailed WB protocol for this protein? I'm using this conditions: 1. boil: 70C, 10 min. 2. Load: 60, 30 and 15 ng of protein (triplicates, because I want to get a standard curve). I load 1 ul per well. 3. Running buffer: 1X MOPS.

Name: Recombinant Human Nrf2 GST (N-Term) Protein
Brand: Novus Biologicals
SKU: H00004780-P01

In regards to your inquiry I have attached the Western Blot conditions that are recommended by Abnova the company that produces this recombinant protein that we distribute. Please see their Western Blot Protocol for full details (http://www.abnova.com/protocol_pdf/Western_Blot.pdf).

Recombinant Human Nrf2 GST (N-Term) Protein

Novus Biologicals, part of Bio-Techne | Catalog # H00004780-P01

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Product Datasheet / COA / SDS

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Product Specifications
Scientific Data
Preparation & Storage
Background
Product Documents
Specific Notices

Key Product Details

Source

Wheat germ

Tag

GST (N-Term)

Conjugate

Unconjugated

Applications

ELISA, Affinity Purification, Enzyme Activity, Microarray, Western Blot

View all Nrf2 Proteins and Enzymes »

Product Specifications

Description

Recombinant protein with GST tag at N-terminal corresponding to the amino acids 1-605 of Human NFE2L2

Source: Wheat Germ (in vitro)

Amino Acid Sequence: MMDLELPPPGLPSQQDMDLIDILWRQDIDLGVSREVFDFSQRRKEYELEKQKKLEKERQEQLQKEQEKAFFAQLQLDEETGEFLPIQPAQHIQSETSGSANYSQVAHIPKSDALYFDDCMQLLAQTFPFVDDNEVSSATFQSLVPDIPGHIESPVFIATNQAQSPETSVAQVAPVDLDGMQQDIEQVWEELLSIPELQCLNIENDKLVETTMVPSPEAKLTEVDNYHFYSSIPSMEKEVGNCSPHFLNAFEDSFSSILSTEDPNQLTVNSLNSDATVNTDFGDEFYSAFIAEPSISNSMPSPATLSHSLSELLNGPIDVSDLSLCKAFNQNHPESTAEFNDSDSGISLNTSPSVASPEHSVESSSYGDTLLGLSDSEVEELDSAPGSVKQNGPKTPVHSSGDMVQPLSPSQGQSTHVHDAQCENTPEKELPVSPGHRKTPFTKDKHSSRLEAHLTRDELRAKALHIPFPVEKIINLPVVDFNEMMSKEQFNEAQLALIRDIRRRGKNKVAAQNCRKRKLENIVELEQDLDHLKDEKEKLLKEKGENDKSLHLLKKQLSTLYLEVFSMLRDEDGKPYSPSEYSLQQTRDGNVFLVPKSKKPDVKKN

Purity

>80% by SDS-PAGE and Coomassie blue staining

Predicted Molecular Mass

94.2 kDa.
Disclaimer note: The observed molecular weight of the protein may vary from the listed predicted molecular weight due to post translational modifications, post translation cleavages, relative charges, and other experimental factors.

Activity

This protein was produced in an in vitro wheat germ expression system that should preserve correct conformational folding that is necessary for biological function. While it is possible that this protein could display some level of activity, the functionality of this protein has not been explicitly measured or validated.

Protein / Peptide Type

Recombinant Protein

Scientific Data Images for Recombinant Human Nrf2 GST (N-Term) Protein

SDS-PAGE: Recombinant Human Nrf2 GST (N-Term) Protein [H00004780-P01]

SDS-Page: Recombinant Human Nrf2 Protein [H00004780-P01] - 12.5% SDS-PAGE Stained with Coomassie Blue.

Formulation, Preparation and Storage

H00004780-P01

Preparation Method	in vitro wheat germ expression system
Formulation	50 mM Tris-HCl, 10 mM reduced Glutathione, pH 8.0 in the elution buffer.
Preservative	No Preservative
Concentration	Please see the vial label for concentration. If unlisted please contact technical services.
Shipping	The product is shipped with dry ice or equivalent. Upon receipt, store it immediately at the temperature recommended below.
Stability & Storage	Store at -80C. Avoid freeze-thaw cycles.

Background: Nrf2

NFE2 (MIM 601490), NFE2L1 (MIM 163260), and NFE2L2 comprise a family of human genes encoding basic leucine zipper (bZIP) transcription factors. They share highly conserved regions that are distinct from other bZIP families, such as JUN (MIM 165160) and FOS (MIM 164810), although remaining regions have diverged considerably from each other (Chan et al., 1995 [PubMed 7868116]).[supplied by OMIM]

Long Name

NF-E2-related Factor 2

Alternate Names

NFE2L2

Gene Symbol