HMGB2 Antibody (JE52-82)

Western Blot: HMGB2 Antibody (JE52-82) [NBP3-32436] - Western blot analysis of HMGB2 on different lysates with Rabbit anti-HMGB2 antibody (NBP3-32436) at 1/1,000 dilution. Lane 1: HeLa cell lysate Lane 2: MCF7 cell lysate Lane 3: HEK-293 cell lysate Lane 4: K-562 cell lysate Lane 5: NIH/3T3 cell lysate Lane 6: PC-12 cell lysate Lane 7:Mouse testis tissue lysate Cell lysates/proteins at 20 ug/Lane. Tissue lysates/proteins at 30 ug/Lane. Predicted band size: 24kDa Observed band size: 28 kDa Exposure time: 3 minutes 10 seconds; 4-20% SDS-PAGE gel. Proteins were transferred to a PVDF membrane and blocked with 5% NFDM/TBST for 1 hour at room temperature. The primary antibody (NBP3-32436) at 1/1,000 dilution was used in 5% NFDM/TBST at 4 overnight. Goat Anti-Rabbit IgG - HRP Secondary Antibody at 1/50,000 dilution was used for 1 hour at room temperature.

Immunohistochemistry: HMGB2 Antibody (JE52-82) [NBP3-32436] - Immunohistochemical analysis of paraffin-embedded human breast carcinoma tissue with Rabbit anti-HMGB2 antibody (NBP3-32436) at 1/500 dilution. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) for 2 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (NBP3-32436) at 1/500 dilution for 1 hour 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.

Immunocytochemistry/ Immunofluorescence: HMGB2 Antibody (JE52-82) [NBP3-32436] - Immunocytochemistry analysis of MCF7 cells labeling HMGB2 with Rabbit anti-HMGB2 antibody (NBP3-32436) at 1/200 dilution. Cells were fixed in 4% paraformaldehyde for 20 minutes at room temperature, permeabilized with 0.1% Triton X-100 in PBS for 5 minutes at room temperature, then blocked with 1% BSA in 10% negative goat serum for 1 hour at room temperature. Cells were then incubated with Rabbit anti-HMGB2 antibody (NBP3-32436) at 1/200 dilution in 1% BSA in PBST overnight at 4 ℃. Goat Anti-Rabbit IgG H&L (iFluor™ 488) was used as the secondary antibody at 1/1,000 dilution. PBS instead of the primary antibody was used as the secondary antibody only control. Nuclear DNA was labelled in blue with DAPI. Beta tubulin (red) was stained at 1/100 dilution overnight at +4℃. Goat Anti-Mouse IgG H&L (iFluor™ 594) was used as the secondary antibody at 1/1,000 dilution.

Flow Cytometry: HMGB2 Antibody (JE52-82) [NBP3-32436] - Flow cytometric analysis of MCF7 cells labeling HMGB2. Cells were fixed and permeabilized. Then stained with the primary antibody (NBP3-32436, 1μg/mL) (red) compared with Rabbit IgG Isotype Control (green). After incubation of the primary antibody at +4℃ for an hour, the cells were stained with a iFluor™ 488 conjugate-Goat anti-Rabbit IgG Secondary antibody at 1/1,000 dilution for 30 minutes at +4℃. Unlabelled sample was used as a control (cells without incubation with primary antibody; black).

• Reactivity: Hu, Mu, Rt
• Applications: WB, Flow, ICC/IF, IHC
• Clone: JE52-82
• Clonality: Monoclonal
• Host: Rabbit
• Conjugate: Unconjugated
• Concentration: 1 mg/ml

HMGB2 Antibody (JE52-82) Summary

• Description: Novus Biologicals Rabbit HMGB2 Antibody (JE52-82) (NBP3-32436) is a recombinant monoclonal antibody validated for use in IHC, WB, Flow and ICC/IF. All Novus Biologicals antibodies are covered by our 100% guarantee.
• Additional Information: Recombinant Monoclonal Antibody
• Immunogen: Synthetic peptide within Human HMGB2 aa 1-50 / 209. (Uniprot: P26583)
• Localization: Chromosome. Cytoplasm. Nucleus.
• Source: HEK293
• Isotype: IgG
• Clonality: Monoclonal
• Host: Rabbit
• Gene: HMGB2
• Purity: Protein A purified

Applications/Dilutions

• Dilutions:
  • Flow Cytometry 1:1000
  • Immunocytochemistry/ Immunofluorescence 1:100-1:250
  • Immunohistochemistry
  • Immunohistochemistry-Paraffin 1:500-1:2000
  • Western Blot 1:1000
• Theoretical MW: 24 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.

Packaging, Storage & Formulations

• Storage: Store at 4C short term. Aliquot and store at -20C long term. Avoid freeze-thaw cycles.
• Buffer: 1*TBS (pH7.4), 0.05% BSA and 40% Glycerol
• Preservative: 0.05% Sodium Azide
• Concentration: 1 mg/ml
• Purity: Protein A purified

Alternate Names for HMGB2 Antibody (JE52-82)

• High mobility group protein 2
• high mobility group protein B2
• high-mobility group box 2
• HMG-2
• HMG2high-mobility group (nonhistone chromosomal) protein 2

Background

HMGB2 encodes a member of the non-histone chromosomal high mobility group protein family. The proteins of this family are chromatin-associated and ubiquitously distributed in the nucleus of higher eukaryotic cells. In vitro studies have demonstrated that this protein is able to efficiently bend DNA and form DNA circles. These studies suggest a role in facilitating cooperative interactions between cis-acting proteins by promoting DNA flexibility. This protein was also reported to be involved in the final ligation step in DNA end-joining processes of DNA double-strand breaks repair and V(D)J recombination.

Limitations

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.

Bioinformatics

• Gene Symbol: HMGB2