PRODUCT CODE: ET1705-86

NRF1 Recombinant Rabbit Monoclonal Antibody [JM89-63] (ET1705-86)

  • Recombinant

Applications

  • WB

  • ICC

  • IF

  • IHC-P

  • FC

  • IP

REACTIVITY

  • Human

  • Mouse

  • Rat

Western blot analysis of NRF1 on Hela cell lysates. Proteins were transferred to a PVDF membrane and blocked with 5% BSA in PBS for 1 hour at room temperature. The primary antibody (ET1705-86, 1/500) was used in 5% BSA at room temperature for 2 hours. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1:5,000 dilution was used for 1 hour at room temperature.
  • Western blot analysis of NRF1 on Hela cell lysates. Proteins were transferred to a PVDF membrane and blocked with 5% BSA in PBS for 1 hour at room temperature. The primary antibody (ET1705-86, 1/500) was used in 5% BSA at room temperature for 2 hours. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1:5,000 dilution was used for 1 hour at room temperature.
  • ICC staining of NRF1 in Hela cells (green). Formalin fixed cells were permeabilized with 0.1% Triton X-100 in TBS for 10 minutes at room temperature and blocked with 1% Blocker BSA for 15 minutes at room temperature. Cells were probed with the primary antibody (ET1705-86, 1/50) for 1 hour at room temperature, washed with PBS. Alexa Fluor®488 Goat anti-Rabbit IgG was used as the secondary antibody at 1/1,000 dilution. The nuclear counter stain is DAPI (blue).
  • ICC staining of NRF1 in MCF-7 cells (green). Formalin fixed cells were permeabilized with 0.1% Triton X-100 in TBS for 10 minutes at room temperature and blocked with 1% Blocker BSA for 15 minutes at room temperature. Cells were probed with the primary antibody (ET1705-86, 1/50) for 1 hour at room temperature, washed with PBS. Alexa Fluor®488 Goat anti-Rabbit IgG was used as the secondary antibody at 1/1,000 dilution. The nuclear counter stain is DAPI (blue).
  • ICC staining of NRF1 in SH-SY5Y cells (green). Formalin fixed cells were permeabilized with 0.1% Triton X-100 in TBS for 10 minutes at room temperature and blocked with 1% Blocker BSA for 15 minutes at room temperature. Cells were probed with the primary antibody (ET1705-86, 1/50) for 1 hour at room temperature, washed with PBS. Alexa Fluor®488 Goat anti-Rabbit IgG was used as the secondary antibody at 1/1,000 dilution. The nuclear counter stain is DAPI (blue).
  • Immunohistochemical analysis of paraffin-embedded rat brain tissue using anti-NRF1 antibody. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) for 20 minutes. The tissues were blocked in 5% BSA for 30 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (ET1705-86, 1/200)  for 30 minutes at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.
  • Immunohistochemical analysis of paraffin-embedded human tonsil tissue using anti-NRF1 antibody. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) for 20 minutes. The tissues were blocked in 5% BSA for 30 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (ET1705-86, 1/200)  for 30 minutes at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.
  • Immunohistochemical analysis of paraffin-embedded human thyroid tissue using anti-NRF1 antibody. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) for 20 minutes. The tissues were blocked in 5% BSA for 30 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (ET1705-86, 1/200)  for 30 minutes at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.
  • Immunohistochemical analysis of paraffin-embedded mouse colon tissue using anti-NRF1 antibody. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) for 20 minutes. The tissues were blocked in 5% BSA for 30 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (ET1705-86, 1/200)  for 30 minutes at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.
  • Flow cytometric analysis of NRF1 was done on 293T cells. The cells were fixed, permeabilized and stained with the primary antibody (ET1705-86, 1/50) (red). After incubation of the primary antibody at room temperature for an hour, the cells were stained with a Alexa Fluor 488-conjugated Goat anti-Rabbit IgG Secondary antibody at 1/1000 dilution for 30 minutes.Unlabelled sample was used as a control (cells without incubation with primary antibody; black).
Western blot analysis of NRF1 on Hela cell lysates. Proteins were transferred to a PVDF membrane and blocked with 5% BSA in PBS for 1 hour at room temperature. The primary antibody (ET1705-86, 1/500) was used in 5% BSA at room temperature for 2 hours. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1:5,000 dilution was used for 1 hour at room temperature.

Applications

  • WB

  • ICC

  • IF

  • IHC-P

  • FC

  • IP

REACTIVITY

  • Human

  • Mouse

  • Rat

SPECIFICATIONS

Product Type

Recombinant Rabbit monoclonal primary

Product Name

NRF1 Recombinant Rabbit Monoclonal Antibody [JM89-63] (ET1705-86)

Immunogen

Synthetic peptide within human nrf1 aa 333-382 / 503.

Host

Rabbit

Positive Control

Hela cell lysates, Hela, MCF-7, SH-SY5Y, rat brain tissue, human tonsil tissue, human thyroid tissue, mouse colon tissue, 293T.

Conjugation

Unconjugated

Clonality

Monoclonal

Clone Number

JM89-63

PROPERTIES

Form

Liquid

Storage Condition

Store at +4C after thawing. Aliquot store at -20C or -80C. Avoid repeated freeze / thaw cycles.

Storage Buffer

1*TBS (pH7.4), 0.05% BSA, 40% Glycerol. Preservative: 0.05% Sodium Azide.

Concentration

1 ug/ul

PURIFICATION

Protein A affinity purified.

MOLECULAR WEIGHT

56 kDa/65kDa(Observed)

Isotype

IgG

APPLICATION DILUTION

  • WB

  • 1:500

  • ICC/IF

  • 1:50-1:200

  • IHC-P

  • 1:50-1:200

  • FC

  • 1:50-1:100

TARGET

UNIPROT #

PROTEIN NAME

NRF1

SYNONYMS

alpha pal antibody; alpha palindromic binding protein antibody; Alpha palindromic-binding protein antibody; Alpha-pal antibody; locus control region factor 1 antibody; NFE2 related factor 1 antibody; NRF-1 antibody; Nrf1 antibody; NRF1_HUMAN antibody; Nuclear respiratory factor 1 antibody

SEQUENCE SIMILARITIES

Belongs to the NRF1/Ewg family.

TISSUE SPECIFICITY

Ubiquitously expressed with strongest expression in skeletal muscle.

POST-TRANSLATIONAL MODIFICATION

Phosphorylation enhances DNA binding.

SUBCELLULAR LOCATION

Nucleus.

FUNCTION

The NF-E2 DNA binding protein is composed of two subunits, p45 and MafK, and it regulates expression of globin genes in developing erythroid cells through interaction with Maf recognition elements (MAREs). A family of NF-E2 related proteins, which are collectively known as the Cap 'n' collar (CNC) family and include Nrf1 (also designated TCF11), Nrf2 and Nrf3, are bZIP transcription factors that heterodimerize with Maf proteins to bind MARE sequences. The Nrf proteins also bind the antioxidant response element (ARE) and are implicated in the regulation of detoxification enzymes and the oxidative stress response. They do so by heterodimerizing with Jun family members (c-Jun, JunB and JunD) to activate gene expression, specifically the detoxifying enzyme, NQO1. Nrf2 is widely expressed and is thought to translocate to the nucleus after treatment with xenobiotics and antioxidants, which stimulate its release from a repressor protein Keap1. Nrf3 is highly expressed in placenta, B cells and monocytes.