High mobility group protein B3 (HMGB3; also HMG-4 and HMG-2a) is a 23 kDa nuclear protein and member of the HMGB family. Human HMGB3 is synthesized as a 200 amino acid (aa) precursor, which is demethylated to produce the 199 aa mature chain. The protein contains two HMG box DNA-binding domains (aa 9‑79 and 93‑161) and an acidic Asp/Glu-rich region (aa 181‑200). Human HMGB3 shares 98% aa sequence identity with mouse and bovine HMGB3. HMGB3 is expressed predominantly in the placenta. It binds preferentially single-stranded DNA and unwinds double stranded DNA.
Key Product Details
Species Reactivity
Human, Mouse
Applications
Western Blot, Immunocytochemistry
Label
Unconjugated
Antibody Source
Monoclonal Mouse IgG1 Clone # 546519
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Product Specifications
Immunogen
E. coli-derived recombinant human HMGB3
Met1-Glu180
Accession # O15347
Met1-Glu180
Accession # O15347
Specificity
Detects human and mouse HMGB3 in direct ELISAs and Western blots.
In direct ELISAs and Western blots, no
cross-reactivity with recombinant human HMG-1, A1A, A1B, or A2 is observed.
Clonality
Monoclonal
Host
Mouse
Isotype
IgG1
Scientific Data Images for Human/Mouse HMGB3 Antibody
Detection of Human HMGB3 by Western Blot.
Western blot shows lysates of human placenta tissue. PVDF Membrane was probed with 2 µg/mL of Human/Mouse HMGB3 Monoclonal Antibody (Catalog # MAB55071) followed by HRP-conjugated Anti-Mouse IgG Secondary Antibody (Catalog # HAF007). A specific band was detected for HMGB3 at approximately 23 kDa (as indicated). This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 1.
HMGB3 in Detroit 551 Human Cell Line.
HMGB3 was detected in immersion fixed Detroit 551 human skin fibroblast cell line using Human/Mouse HMGB3 Monoclonal Antibody (Catalog # MAB55071) at 10 µg/mL for 3 hours at room temperature. Cells were stained using the NorthernLights™ 557-conjugated Anti-Mouse IgG Secondary Antibody (red, upper panel; Catalog # NL007) and counterstained with DAPI (blue, lower panel). Specific staining was localized to nuclei. View our protocol for Fluorescent ICC Staining of Cells on Coverslips.
Applications for Human/Mouse HMGB3 Antibody
Application
Recommended Usage
Immunocytochemistry
8-25 µg/mL
Sample: Immersion fixed Detroit 551 human skin fibroblast cell line
Sample: Immersion fixed Detroit 551 human skin fibroblast cell line
Western Blot
2 µg/mL
Sample: Human placenta tissue
Sample: Human placenta tissue
Formulation, Preparation, and Storage
Purification
Protein A or G purified from hybridoma culture supernatant
Reconstitution
Sterile PBS to a final concentration of 0.5 mg/mL. For liquid material, refer to CoA for concentration.
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Formulation
Lyophilized from a 0.2 μm filtered solution in PBS with Trehalose. *Small pack size (SP) is supplied either lyophilized or as a 0.2 µm filtered solution in PBS.
Shipping
Lyophilized product is shipped at ambient temperature. Liquid small pack size (-SP) is shipped with polar packs. Upon receipt, store immediately at the temperature recommended below.
Stability & Storage
Use a manual defrost freezer and avoid repeated freeze-thaw cycles.
- 12 months from date of receipt, -20 to -70 °C as supplied.
- 1 month, 2 to 8 °C under sterile conditions after reconstitution.
- 6 months, -20 to -70 °C under sterile conditions after reconstitution.
Calculators
Background: HMGB3
Long Name
High Mobility Group Box 3
Alternate Names
HMG2A, HMG4
Gene Symbol
HMGB3
UniProt
Additional HMGB3 Products
Product Documents for Human/Mouse HMGB3 Antibody
Product Specific Notices for Human/Mouse HMGB3 Antibody
For research use only
Related Research Areas
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Protocols
Find general support by application which include: protocols, troubleshooting, illustrated assays, videos and webinars.
- Appropriate Fixation of IHC/ICC Samples
- Cellular Response to Hypoxia Protocols
- Detection & Visualization of Antibody Binding
- ICC Cell Smear Protocol for Suspension Cells
- ICC Immunocytochemistry Protocol Videos
- ICC for Adherent Cells
- Immunocytochemistry (ICC) Protocol
- Immunocytochemistry Troubleshooting
- Immunofluorescence of Organoids Embedded in Cultrex Basement Membrane Extract
- Immunohistochemistry (IHC) and Immunocytochemistry (ICC) Protocols
- Preparing Samples for IHC/ICC Experiments
- Preventing Non-Specific Staining (Non-Specific Binding)
- Primary Antibody Selection & Optimization
- Protocol for VisUCyte™ HRP Polymer Detection Reagent
- Protocol for the Fluorescent ICC Staining of Cell Smears - Graphic
- Protocol for the Fluorescent ICC Staining of Cultured Cells on Coverslips - Graphic
- Protocol for the Preparation and Fluorescent ICC Staining of Cells on Coverslips
- Protocol for the Preparation and Fluorescent ICC Staining of Non-adherent Cells
- Protocol for the Preparation and Fluorescent ICC Staining of Stem Cells on Coverslips
- Protocol for the Preparation of a Cell Smear for Non-adherent Cell ICC - Graphic
- R&D Systems Quality Control Western Blot Protocol
- TUNEL and Active Caspase-3 Detection by IHC/ICC Protocol
- The Importance of IHC/ICC Controls
- Troubleshooting Guide: Western Blot Figures
- Western Blot Conditions
- Western Blot Protocol
- Western Blot Protocol for Cell Lysates
- Western Blot Troubleshooting
- Western Blot Troubleshooting Guide
- View all Protocols, Troubleshooting, Illustrated assays and Webinars