Key Product Details

Species Reactivity

Validated:

Human, Mouse

Cited:

Human, Mouse

Applications

Validated:

Western Blot, Immunocytochemistry/ Immunofluorescence

Cited:

Western Blot

Label

Unconjugated

Antibody Source

Polyclonal Rabbit IgG

Format

BSA Free
View all FXR/NR1H4 Primary Antibodies »

Product Specifications

Immunogen

A synthetic peptide made to the C-terminus of the human FXR protein. [UniProt# Q96RI1]

Reactivity Notes

Mouse reactivity reported in scientific literature (PMID: 29491146).

Localization

Cytoplasmic, nuclear

Clonality

Polyclonal

Host

Rabbit

Isotype

IgG

Theoretical MW

66 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.

Description

Novus Biologicals Rabbit FXR/NR1H4 Antibody - BSA Free (NB400-153) is a polyclonal antibody validated for use in WB and ICC/IF. Anti-FXR/NR1H4 Antibody: Cited in 4 publications. All Novus Biologicals antibodies are covered by our 100% guarantee.

Scientific Data Images for FXR/NR1H4 Antibody - BSA Free

Western Blot: FXR/NR1H4 Antibody [NB400-153]

Western Blot: FXR/NR1H4 Antibody [NB400-153]

Western Blot: FXR/NR1H4 Antibody [NB400-153] - Total protein from human A549, U2OS cell and mouse 3T3 cells was separated on a 7.5% gel by SDS-PAGE, transferred to PVDF membrane and blocked in 5% non-fat milk in TBST. The membrane was probed with 2.0 ug/ml anti-FXR (NB400-153) in block buffer and detected with an anti-rabbit HRP secondary antibody using NovaLume chemiluminescence detection reagent (NPB2-61915).
Immunocytochemistry/ Immunofluorescence: FXR/NR1H4 Antibody [NB400-153]

Immunocytochemistry/ Immunofluorescence: FXR/NR1H4 Antibody [NB400-153]

Immunocytochemistry/Immunofluorescence: FXR/NR1H4 Antibody [NB400-153] - A549 cells were fixed for 10 minutes using 10% formalin and then permeabilized for 5 minutes using 1X PBS + 0.5% Triton-X100. The cells were incubated with anti-FXR/NR1H4 at 2 ug/ml overnight at 4C and detected with an anti-rabbit Dylight 488 (Green) at a 1:500 dilution. Alpha tubulin (DM1A) NB100-690 was used as a co-stain at a 1:1000 dilution and detected with an anti-mouse Dylight 550 (Red) at a 1:500 dilution. Nuclei were counterstained with DAPI (Blue). Cells were imaged using a 40X objective.
Western Blot: FXR/NR1H4 Antibody [NB400-153]

Western Blot: FXR/NR1H4 Antibody [NB400-153]

Western Blot: FXR Antibody [NB400-153] - Detection of FXR in HeLa nuclear extract using NB400-153. ECL 30 minute detection.
Immunocytochemistry/ Immunofluorescence: FXR/NR1H4 Antibody [NB400-153]

Immunocytochemistry/ Immunofluorescence: FXR/NR1H4 Antibody [NB400-153]

Immunocytochemistry/Immunofluorescence: FXR/NR1H4 Antibody [NB400-153] - NIH3T3 cells were fixed for 10 minutes using 10% formalin and then permeabilized for 5 minutes using 1X PBS + 0.5% Triton-X100. The cells were incubated with anti-FXR/NR1H4 at 2 ug/ml overnight at 4C and detected with an anti-rabbit Dylight 488 (Green) at a 1:500 dilution. Alpha tubulin (DM1A) NB100-690 was used as a co-stain at a 1:1000 dilution and detected with an anti-mouse Dylight 550 (Red) at a 1:500 dilution. Nuclei were counterstained with DAPI (Blue). Cells were imaged using a 40X objective.

Applications for FXR/NR1H4 Antibody - BSA Free

Application
Recommended Usage

Immunocytochemistry/ Immunofluorescence

1:100

Western Blot

1 - 4 ug/mL
Application Notes
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.

Reviewed Applications

Read 4 reviews rated 4.5 using NB400-153 in the following applications:

Formulation, Preparation, and Storage

Purification

Immunogen affinity purified

Formulation

Tris-Glycine, 0.15M NaCl

Format

BSA Free

Preservative

0.05% Sodium Azide

Concentration

1 mg/ml

Shipping

The product is shipped with polar packs. Upon receipt, store it immediately at the temperature recommended below.

Stability & Storage

Store at 4C short term. Aliquot and store at -20C long term. Avoid freeze-thaw cycles.

Background: FXR/NR1H4

FXR (farnesoid X receptor; NR1H4) belongs to nuclear receptor superfamily of ligand-activated transcription factors (NR1 subfamily) and acts as a receptor for bile acids such as chenodeoxycholic acid, lithocholic acid and deoxycholic acid. FXR is mainly expressed in liver, intestine, kidney, and adrenal glands. FXR heterodimerizes with retinoid X receptor (RXR alpha/NR2B1), associates with FXREs (FXR response elements) that are usually but not exclusively inverted repeat-1 (IR-1), and regulates the transcription of its target genes. FXR interacts with CARM1, SMARD1 and after activation by agonist binding, it interacts with a coactivator, NCOA1 or NCOA2. FXR has been shown to repress transcription of CYP7A1 (cholesterol 7-alpha-hydroxylase gene) through induction of NR0B2 or FGF19 expression, via two distinct mechanisms. FXR activates the intestinal bile acid-binding protein (IBABP) and transcription of bile salt export pump ABCB11 by directly recruiting histone methyltransferase CARM1 to this locus. Altogether, FXR is critical for maintaining a homeostasis in bile acid, cholesterol, lipoprotein, and triglyceride metabolism.

Long Name

Farnesoid X-activated receptor

Alternate Names

BAR, HRR-1, NR1H4, RIP14

Gene Symbol

NR1H4

Additional FXR/NR1H4 Products

Product Documents for FXR/NR1H4 Antibody - BSA Free

Certificate of Analysis

To download a Certificate of Analysis, please enter a lot or batch number in the search box below.

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Product Specific Notices for FXR/NR1H4 Antibody - BSA Free

This product is for research use only and is not approved for use in humans or in clinical diagnosis. Primary Antibodies are guaranteed for 1 year from date of receipt.

Citations for FXR/NR1H4 Antibody - BSA Free

Customer Reviews for FXR/NR1H4 Antibody - BSA Free (4)

4.5 out of 5
4 Customer Ratings
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4 Stars
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Showing  1 - 4 of 4 reviews Showing All
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  • FXR/NR1H4 Antibody
    Name: Anonymous
    Application: Western Blot
    Sample Tested: HepG2 liver cells and Huh-7 human hepatoma cell line
    Species: Human
    Verified Customer | Posted 12/18/2023
    It works well for the WB analysis
    FXR/NR1H4 Antibody - BSA Free NB400-153
  • FXR/NR1H4 Antibody
    Name: Anonymous
    Application: Immunocytochemistry
    Sample Tested: hela cell
    Species: Human
    Verified Customer | Posted 04/04/2020
    FXR/NR1H4 Antibody - BSA Free NB400-153
  • FXR/NR1H4 Antibody
    Name: Anonymous
    Application: Western Blot
    Sample Tested: Hela whole cell lysate
    Species: Human
    Verified Customer | Posted 12/27/2019
    FXR/NR1H4 Antibody - BSA Free NB400-153
  • FXR/NR1H4 Antibody
    Name: Anonymous
    Application: Western Blot
    Sample Tested: HepG2 and Human liver cancer cell line (HuH7)
    Species: Human
    Verified Customer | Posted 02/19/2019
    FXR/NR1H4 Antibody - BSA Free NB400-153

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Protocols

View specific protocols for FXR/NR1H4 Antibody - BSA Free (NB400-153):

FXR/NR1H4 Antibody:
Immunocytochemistry Protocol

Culture cells to appropriate density in 35 mm culture dishes or 6-well plates.

1. Remove culture medium and add 10% formalin to the dish. Fix at room temperature for 30 minutes.
2. Remove the formalin and add ice cold methanol. Incubate for 5-10 minutes.
3. Remove methanol and add washing solution (i.e. PBS). Be sure to not let the specimen dry out. Wash three times for 10 minutes.
4. To block nonspecific antibody binding incubate in 10% normal goat serum from 1 hour to overnight at room temperature.
5. Add primary antibody at appropriate dilution and incubate at room temperature from 2 hours to overnight at room temperature.
6. Remove primary antibody and replace with washing solution. Wash three times for 10 minutes.
7. Add secondary antibody at appropriate dilution. Incubate for 1 hour at room temperature.
8. Remove antibody and replace with wash solution, then wash for 10 minutes. Add Hoechst 33258 to wash solution at 1:25,0000 and incubate for 10 minutes. Wash a third time for 10 minutes.
9. Cells can be viewed directly after washing. The plates can also be stored in PBS containing Azide covered in Parafilm (TM). Cells can also be cover-slipped using Fluoromount, with appropriate sealing.

*The above information is only intended as a guide. The researcher should determine what protocol best meets their needs. Please follow safe laboratory procedures.

FXR/NR1H4 Antibody:
Western Blot Protocol
1. Perform SDS-PAGE (4-12%) on samples to be analyzed, loading 20 ug of total transfected protein per lane.
2. Transfer proteins to Nitrocellulose according to the instructions provided by the manufacturer of the transfer
apparatus.
3. Stain the blot using ponceau S for 1-2 minutes to access the transfer of proteins onto the nitrocellulose membrane. Rinse the blot in water to remove excess stain and mark the lane locations and locations of molecular weight markers using a pencil.
4. Rinse the blot in TBS for approximately 5 minutes.
5. Block the membrane using 5% non-fat dry milk in TBS + 0.5% BSA for 1 hour.
6. Dilute the rabbit anti-FXR primary antibody (NB 400-153) in blocking buffer and incubate 2 hours at room temperature.
7. Wash the membrane in water for 5 minutes and apply the diluted rabbit-IgG HRP-conjugated secondary antibody in blocking buffer (as per manufacturers instructions) and incubate 1 hour at room temperature.
8. Wash the blot in TBS containing 0.05-0.1% Tween-20 for 10-20 minutes.
9. Wash the blot in type I water for an additional 10-20 minutes (this step can be repeated as required to reduce background).
10. Apply the detection reagent of choice in accordance with the manufacturers instructions (Amersham's ECL is the standard reagent used at Novus Biologicals).
Note: Tween-20 can be added to the blocking buffer at a final concentration of 0.05-0.2%, provided it does not interfere with antibody-antigen binding.

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