HMGB1 Recombinant antibody

• Featured Product
• KD/KO Validated

HMGB1 Uni-rAb^TM Recombinant Antibody for WB, IHC, IF/ICC, FC (Intra), ELISA

Host / Isotype

Rabbit / IgG

Reactivity

human, mouse, rat

Applications

WB, IHC, IF/ICC, FC (Intra), ELISA

Conjugate

Unconjugated

CloneNo.

230182G6

Cat no : 82973-1-RR

Synonyms

HMG1, HMG 1, High mobility group protein B1, High mobility group protein 1, high mobility group box 1

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Validation Data Gallery

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  WB analysis using 82973-1-RR

  Various lysates were subjected to SDS PAGE followed by western blot with 82973-1-RR (HMGB1 antibody) at dilution of 1:10000 incubated at room temperature for 1.5 hours.

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  WB analysis of HeLa using 82973-1-RR

  WB result of HMGB1 antibody (82973-1-RR; 1:3000; incubated at room temperature for 1.5 hours) with negative control and HMGB1 knockout HeLa cells.

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  IHC staining of mouse brain using 82973-1-RR

  Immunohistochemical analysis of paraffin-embedded mouse brain tissue slide using 82973-1-RR (HMGB1 antibody) at dilution of 1:1000 (under 40x lens). Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0).

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  IHC staining of mouse brain using 82973-1-RR

  Immunohistochemical analysis of paraffin-embedded mouse brain tissue slide using 82973-1-RR (HMGB1 antibody) at dilution of 1:1000 (under 10x lens). Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0).

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  IHC staining of human normal colon using 82973-1-RR

  Immunohistochemical analysis of paraffin-embedded human colon tissue slide using 82973-1-RR (HMGB1 antibody) at dilution of 1:2000 (under 20x lens). Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0).

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  IHC staining of human normal colon using 82973-1-RR

  Immunohistochemical analysis of paraffin-embedded human colon tissue slide using 82973-1-RR (HMGB1 antibody) at dilution of 1:2000 (under 20x lens). Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0).

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  IF Staining of HeLa using 82973-1-RR

  Immunofluorescent analysis of (4% PFA) fixed HeLa cells using HMGB1 antibody (82973-1-RR, Clone: 230182G6 ) at dilution of 1:250 and CoraLite®488-Conjugated AffiniPure Goat Anti-Rabbit IgG(H+L) (SA00013-2), CL594-Phalloidin (red).

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  IF Staining of HeLa using 82973-1-RR

  Immunofluorescent analysis of (4% PFA) fixed HeLa cells using HMGB1 antibody (82973-1-RR, Clone: 230182G6 ) at dilution of 1:250 and CoraLite®488-Conjugated AffiniPure Goat Anti-Rabbit IgG(H+L) (SA00013-2), CL594-Phalloidin (red).

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  FC experiment of HeLa using 82973-1-RR

  1x10^6 HeLa cells were intracellularly stained with 0.4 ug HMGB1 Recombinant antibody (82973-1-RR, Clone:230182G6) and CoraLite®488-Conjugated AffiniPure Goat Anti-Rabbit IgG(H+L) (SA00013-2)(red), or 0.4 ug Isotype Control (blue). Cells were fixed and permeabilized with Transcription Factor Staining Buffer Kit (PF00011).

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  Affinity and Kinetic Characterization of 82973-1-RR

  Biolayer interferometry (BLl) kinetic assays of 82973-1-RR against Human HMGB1 were performed. The affinity constant is 0.246 nM.

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Tested Applications

• Positive WB detected in: L02 cells, HeLa cells, Jurkat cells, HEK-293 cells, K-562 cells, HCT 116 cells, mouse brain tissue, rat brain tissue
• Positive IHC detected in: human colon tissue, mouse brain tissue; Note: suggested antigen retrieval with TE buffer pH 9.0; (*) Alternatively, antigen retrieval may be performed with citrate buffer pH 6.0
• Positive IF/ICC detected in: HeLa cells
• Positive FC (Intra) detected in: HeLa cells

Recommended dilution

• Western Blot (WB): WB : 1:5000-1:50000
• Immunohistochemistry (IHC): IHC : 1:1000-1:4000
• Immunofluorescence (IF)/ICC: IF/ICC : 1:125-1:500
• Flow Cytometry (FC) (INTRA): FC (INTRA) : 0.40 ug per 10^6 cells in a 100 µl suspension

It is recommended that this reagent should be titrated in each testing system to obtain optimal results.

Sample-dependent, Check data in validation data gallery.

Product Information

82973-1-RR targets HMGB1 in WB, IHC, IF/ICC, FC (Intra), ELISA applications and shows reactivity with human, mouse, rat samples.

• Tested Reactivity: human, mouse, rat
• Host / Isotype: Rabbit / IgG
• Class: Recombinant
• Type: Antibody
• Immunogen: HMGB1 fusion protein Ag1264
• Full Name: high-mobility group box 1
• Calculated Molecular Weight: 25 kDa
• Observed Molecular Weight: 25-30 kDa
• GenBank Accession Number: BC003378
• Gene Symbol: HMGB1
• Gene ID (NCBI): 3146
• Conjugate: Unconjugated
• Form: Liquid
• Purification Method: Protein A purification
• Storage Buffer: PBS with 0.02% sodium azide and 50% glycerol pH 7.3.
• Storage Conditions: Store at -20°C. Stable for one year after shipment. Aliquoting is unnecessary for -20^oC storage. 20ul sizes contain 0.1% BSA.

Background Information

The HMG (high mobility group) proteins are nonhistone chromosomal proteins that is present in almost all eukaryotic cells, and it functions to stabilize NUCLEOSOME formation and acts as a transcription-factor-like protein that regulates the expression of several genes[PMID: 18160415]. Once injury, infection or other inflammatory stimuli, activated macrophages, mature dendritic cells and natural killer cells can secret HMGB1, which act as a crucial cytokine[PMID: 20163887]. HMGB1 also involved in V(D)J recombination by acting as a cofactor of the RAG complex, stimulating cleavage and RAG protein binding at the 23 bp spacer of conserved recombination signal sequences (RSS)[PMID: 19360789 ]. Act as a Heparin-binding protein that has a role in the extension of neurite-type cytoplasmic processes in developing cells. HMGB1 (high mobility group box 1) modulates gene expression in the nucleus, but certain immune cells secrete HMGB1 as an extracellular Alarmin to signal tissue damage.The nuclear HMGB1 relocalizes to the extracellular milieu in senescent human and mouse cells in culture and in vivo, which stimulated cytokine secretion through TLR-4 signaling (23649808).