Key Product Details

Validated by

Biological Validation

Species Reactivity

Validated:

Human, Mouse

Cited:

Human, Mouse

Predicted:

Primate (94%), Rat (94%). Backed by our 100% Guarantee.

Applications

Validated:

Immunohistochemistry, Immunohistochemistry-Paraffin, Western Blot, Immunocytochemistry/ Immunofluorescence, Gel Super Shift Assays, Gel Supershift Assay

Cited:

Western Blot, Gel Supershift Assay

Label

Unconjugated

Antibody Source

Polyclonal Rabbit IgG

Format

BSA Free
View all IRP2 Primary Antibodies »

Product Specifications

Immunogen

A synthetic peptide made to an internal portion of the murine IRP2 protein sequence (between residues 100-200). [UniProt Q811J3]

Reactivity Notes

Preliminary results have shown poor reactivity against human protein. Immunogen displays the following percentage of sequence identity for non-tested species: primate (94%) and rat (94%) proteins. Human reactivity reported in scientific literature (PMID: 28888202).

Localization

Cytoplasmic

Clonality

Polyclonal

Host

Rabbit

Isotype

IgG

Scientific Data Images for IRP2 Antibody - BSA Free

Western Blot: IRP2 Antibody [NB100-1798]

Western Blot: IRP2 Antibody [NB100-1798]

Western Blot: IRP2 Antibody [NB100-1798] - Raw macrophages (mouse) were either left untreated (lane 1) or treated overnight with 100mM DFO (lane 2) or 100mM hemin (lane 3). Proteins extracts were subjected to western blotting using IRP2 or b-actin antibodies diluted 1:500 in TBS-Tween.
Immunohistochemistry-Paraffin: IRP2 Antibody [NB100-1798]

Immunohistochemistry-Paraffin: IRP2 Antibody [NB100-1798]

Immunohistochemistry-Paraffin: IRP2 Antibody [NB100-1798] - IHC analysis of a formalin fixed paraffin-embedded (FFPE) human placenta using 1:300 conc. of IRP2 antibody on a Bond Rx autostainer (Leica Biosystems). The assay involved 30 minutes of heat induced antigen retrieval (HIER) using 10mM sodium citrate buffer (pH 9.0) and endogenous peroxidase quenching with peroxide block. The sections were incubated with primary antibody for 15 minutes and Bond Polymer Refine Detection (Leica Biosystems) with DAB was used for signal development followed by counterstaining with hematoxylin. Cytoplasmic staining was observed in villi of endothelial cells.
Western Blot: IRP2 Antibody [NB100-1798]

Western Blot: IRP2 Antibody [NB100-1798]

Western Blot: IRP2 Antibody [NB100-1798] - Detection of IRP2 in mouse liver lysate.
Western Blot: IRP2 Antibody [NB100-1798]

Western Blot: IRP2 Antibody [NB100-1798]

Western Blot: IRP2 Antibody [NB100-1798] - Detection of IRP2 in RAW 264.7 lysate using NB100-1798. Lane 1: Lot B Lane 2: Lot C

Applications for IRP2 Antibody - BSA Free

Application
Recommended Usage

Gel Super Shift Assays

reported in scientific literature (PMID 26752519)

Immunocytochemistry/ Immunofluorescence

1:500

Immunohistochemistry

1:200-1:500

Immunohistochemistry-Paraffin

1:200-1:500

Western Blot

1:500-1:1000

Formulation, Preparation, and Storage

Purification

Immunogen affinity purified

Formulation

Tris-Glycine and 0.15M NaCl

Format

BSA Free

Preservative

0.05% Sodium Azide

Concentration

1.15 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: IRP2

Iron regulatory proteins -1 and -2 (IRP1, IRP2) are homologous cytoplasmic RNA-binding proteins that controls cellular iron metabolism and maintain systemic iron homeostasis in vertebrates along with serving some additional iron-independent functions. IRP2 belongs to aconitase/IPM isomerase family and it binds to IRES (iron-responsive elements), which are stem-loop structures found in 5'-UTR of ferritin, and delta aminolevulinic acid synthase mRNAs, and in the 3'-UTR of transferrin receptor mRNA. IRP2-IRE binding results in the repression of its mRNA translation and its binding to transferrin receptor mRNA inhibits the degradation of this otherwise quickly degradable mRNA. IRP2 is a substrate of FBXL5, an F-box protein that assembles together with Skp1, Cul1 and Rbx1 into an E3 ubiquitin ligase complex. When iron/O2 are high, IRP2 gets ubiquitinated/degraded by SCF complex containing FBXL5, whereas under conditions of low iron/O2, FBXL5 is degraded preventing IRP2 ubiquitination leading to its RNA-binding activity. IRP2 deficient mice depicts microcytosis, misregulated iron metabolism (in CNS, intestine as well as liver associated with FTH1 and FTL up-regulation) and iron deficiency in splenic macrophages associated with down-regulation of FTH1, FTL, and FPN1.

Alternate Names

ACO3, EC 4.2.1.3, FLJ23381, IRE-BP 2, Iron regulatory protein 2, iron-responsive element binding protein 2, iron-responsive element-binding protein 2, IRP2IRP2AD

Entrez Gene IDs

3658 (Human); 64602 (Mouse)

Gene Symbol

IREB2

UniProt

Q811J3 (Mouse)

Additional IRP2 Products

Product Documents for IRP2 Antibody - BSA Free

Certificate of Analysis

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Product Specific Notices for IRP2 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 IRP2 Antibody - BSA Free

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Protocols

View specific protocols for IRP2 Antibody - BSA Free (NB100-1798):

IRP2 Antibody:
Western Blot Protocol

1. Perform SDS-PAGE (4-12%) on samples to be analyzed, loading 50 ug of total protein per lane.
2. Transfer proteins to Nitrocellulose according to the instructions provided by the manufacturer of the transfer apparatus.
3. Rinse membrane with dH2O and then 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 + 1% BSA in TBS for 2 hours at room temperature.
6. Rinse the membrane in dH2O and then wash the membrane in wash buffer [TBS + 0.1% Tween] 3 times for 10 minutes each.
7. Dilute the rabbit anti-IRP2 primary antibody (NB 100-1798) in blocking buffer and incubate 1 hour at room temperature.
8. Rinse the membrane in dH2O and then wash the membrane in wash buffer [TBS + 0.1% Tween] 3 times for 10 minutes each.
9. Apply the diluted rabbit-IgG HRP-conjugated secondary antibody in blocking buffer (as per manufacturers instructions) and incubate 1 hour at room temperature.
10. Wash the blot in wash buffer [TBS + 0.1% Tween] 3 times for 10 minutes each (this step can be repeated as required to reduce background).
11. Apply the detection reagent of choice in accordance with the manufacturers instructions (Pierce's ECL).

Note: Tween-20 can be added to the blocking or antibody diultion buffer at a final concentration of 0.05-0.2%, provided it does not interfere with antibody-antigen binding.

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FAQs

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