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Key Product Details

Species Reactivity

Human, Mouse

Applications

Flow (Intracellular), Flow Cytometry, Immunocytochemistry/ Immunofluorescence, Immunohistochemistry, Immunohistochemistry-Frozen, Immunohistochemistry-Paraffin, Immunoprecipitation, Western Blot

Label

Unconjugated

Antibody Source

Monoclonal Mouse IgG1 Clone # 2D9

Format

BSA Free

Concentration

1.0 mg/ml

Product Summary for NOD2 Antibody (2D9) - BSA Free

Immunogen

Recombinant human NOD2 protein corresponding to residues 28-301. [UniProt# Q9HC29]

Predicted Species

Primate (92%). Backed by our 100% Guarantee.

Localization

Cytoplasmic

Clonality

Monoclonal

Host

Mouse

Isotype

IgG1

Theoretical MW

110 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.

Scientific Data Images for NOD2 Antibody (2D9) - BSA Free

Western Blot: NOD2 Antibody (2D9) [NB100-524] - HCMV infection induces NOD2 mRNA and protein in HFFs and U373 cells. E. U373 glioma cells were infected with HCMV Towne strain and levels of NOD1, NOD2 and GAPDH mRNAs were measured by qRT-PCR at indicated time points. F. HFFs were infected with HCMV (Towne) at MOI of 1 PFU/cell and levels of NOD2 protein and B-actin were determined 48 and 72 hpi. G. HFFs were infected with HCMV (Towne) strain at MOI of 0.03 or 3 PFU/cell and levels of NOD2 protein and B-actin were determined at 48 hpi. Quantitative data represent mean values (+/-SD) of triplicate determinations from three independent experiments (*p<0.05, **p<0.01, ***p<0.001, one-way ANOVA test). Image collected and cropped by CiteAb from the following publication (//doi.org/10.1371/journal.pone.0092704.g001) licensed under a CC-BY license.
Immunohistochemistry-Frozen: NOD2 Antibody (2D9) [NB100-524] - Overlay of NOD2-DyLight 488 (green) with phase contrast of murine colon. Image from verified customer review.
Flow (Intracellular): NOD2 Antibody (2D9) [NB100-524] - An intracellular stain was performed on THP-1 cells with NOD2 (2D9) antibody NB100-524AF488 (blue) and a matched isotype control NBP2-27287AF488 (orange). Cells were fixed with 4% PFA and then permeablized with 0.1% saponin. Cells were incubated in an antibody dilution of 5 ug/mL for 30 minutes.

Applications for NOD2 Antibody (2D9) - BSA Free

Application
Recommended Usage

Flow Cytometry

reported in scientific literature (PMID 28241127)

Immunocytochemistry/ Immunofluorescence

1:50-1:500

Immunohistochemistry

1:10-1:500

Immunohistochemistry-Paraffin

1:10-1:500

Immunoprecipitation

1:10-1:500

Western Blot

1:100-1:2000
Application Notes
In WB, this antibody has been tested with NOD2 transfected 293T lysates and HT29 human colorectal adenocarcinoma lysates (endogenous detection has some background) where a band is seen at ~110 kDa. 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.
Please Note: Optimal dilutions of this antibody should be experimentally determined.

Reviewed Applications

Read 1 review rated 5 using NB100-524 in the following applications:

Published Applications

Read 26 publications using NB100-524 in the following applications:

Formulation, Preparation, and Storage

Purification

Protein G purified

Formulation

PBS

Format

BSA Free

Preservative

0.05% Sodium Azide

Concentration

1.0 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: NOD2

The NOD2 (nucleotide-binding oligomerization domain containing 2) protein is expressed in the antigen-presenting cells (monocytes and macrophages) and in intestinal Paneth cells, keratinocytes, T cells, NK cells, and CD34+ stem cells. Alongwith Apaf-1 and CIITA, NOD2 belongs to NLR/NOD leucine-rich repeat containing protein family. NOD2 generally localizes to the cytoplasm from where it translocates to cell membranes, a mechanism suggested as important for its functionality. After interacting with CARD-CARD, NOD2 may bind/activate RIP2/RICK and this active NOD2:RIP2 complex then stimulates MAPK (JNK, ERK, p38), NF-kB, autophagy and antigen presentation. NOD2 has an ability to senses the presence of MDP (muramyl dipeptide), a breakdown product of peptidoglycan found in bacteria, and after recognition of intracellular bacteria, NOD2 pathway execute directing the secretion of antimicrobial peptides and cytokines/chemokines to induce innate immune clearance of the pathogen as well as to tailor the adaptive immune system to help fight the bacterial infection. Defects in NOD2 are the cause of Blau syndrome (BS), susceptibility to inflammatory bowel disease type 1 (IBD1) and sarcoidosis early-onset (EOS).

Alternate Names

BLAU, CARD15caspase recruitment domain protein 15, caspase recruitment domain family, member 15, Caspase recruitment domain-containing protein 15, CDACUG, CLR16.3, IBD1NLR family, CARD domain containing 2, Inflammatory bowel disease protein 1, NLRC2, NOD-like receptor C2, nucleotide-binding oligomerization domain 2, nucleotide-binding oligomerization domain containing 2, nucleotide-binding oligomerization domain, leucine rich repeat and CARD domaincontaining 2, nucleotide-binding oligomerization domain-containing protein 2, PSORAS1NOD2B

Gene Symbol

NOD2

Product Documents for NOD2 Antibody (2D9) - BSA Free

Certificate of Analysis

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

Product Specific Notices for NOD2 Antibody (2D9) - 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.

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Protocols

View specific protocols for NOD2 Antibody (2D9) - BSA Free (NB100-524):

[[URL:https://www.novusbio.com/products/nod2-antibody-2d9_nb100-524]][[Captio… Antibody (2D9)]]
Western Blot Protocol

1. Perform SDS-PAGE (3-8%) on samples to be analyzed, loading 20ug (transfected lysates) or 50ug (endogenous) of total 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 mouse anti-NOD2 primary antibody (NB 100-524) in blocking buffer and incubate 2 hours at room temperature.

7. Wash the membrane in water for 5 minutes and apply the diluted mouse-IgG HRP-conjugated secondary antibody in blocking buffer (as per manufacturer's 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 manufacturer's 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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