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Home > Product > Antibody > Rabbit Anti-HMGB1 antibody
High mobility group protein B1; Amphoterin; High mobility group 1; High Mobility Group Box 1; High mobility group protein 1; HMG3; HMGB 1; HMGB-1; Hmgb1 protein; Nonhistone chromosomal protein HMG1; SBP 1; SBP-1; Sulfoglucuronyl carbohydrate binding prote
Cat:
SL0664R
Species Reactivity：
Human,Mouse,Rat,(predicted: Cow,)
Immunogen：
KLH conjugated synthetic peptide derived from human HMGB1:75-170/215
Format：
Liquid
Storage instructions：
Shipped at 4℃. Store at -20 °C for one year. Avoid repeated freeze/thaw cycles.
Concentration：
1mg/ml
Clonality：
Polyclonal
Isotype：
IgG
Applications：
WB=1:500-2000ELISA=1:5000-10000IHC-P=1:100-500IHC-F=1:100-500Flow-Cyt=1μg/TestICC=1:100IF=1:100-500（Paraffin sections need to do antigen repair）not yet tested in other applications.optimal dilutions/concentrations should be determined by the end user.
Host：
Rabbit
Product Overview：
Sample: bone (mouse) Lysate at 40 ugPrimary: Anti- HMGB1(SL0664R)at 1/300 dilutionSecondary: IRDye800CW Goat Anti-Rabbit IgG at 1/20000 dilutionPredicted band size: 25 kDObserved band size: 27kDParaformaldehyde-fixed, paraffin embedded (Rat stomach); Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15min; Block endogenous peroxidase by 3% hydrogen peroxide for 20 minutes; Blocking buffer (normal goat serum) at 37°C for 30min; Antibody incubation with (HMGB1) Polyclonal Antibody, Unconjugated (SL0664R) at 1:400 overnight at 4°C, followed by operating according to SP Kit(Rabbit) (sp-0023) instructions and DAB staining.Tissue/cell: human endometrium carcinoma; 4% Paraformaldehyde-fixed and paraffin-embedded; Antigen retrieval: citrate buffer ( 0.01M, pH 6.0 ), Boiling bathing for 15min; Block endogenous peroxidase by 3% Hydrogen peroxide for 30min; Blocking buffer (normal goat serum,SLC0005) at 37℃ for 20 min; Incubation: Anti-HMGB1 Polyclonal Antibody, Unconjugated(SL0664R) 1:200, overnight at 4癈, followed by conjugation to the secondary antibody(SP-0023) and DAB(SLC0010) stainingTissue/cell: rat liver tissue; 4% Paraformaldehyde-fixed and paraffin-embedded; Antigen retrieval: citrate buffer ( 0.01M, pH 6.0 ), Boiling bathing for 15min; Block endogenous peroxidase by 3% Hydrogen peroxide for 30min; Blocking buffer (normal goat serum,SLC0005) at 37℃ for 20 min; Incubation: Anti-HMGB1 Polyclonal Antibody, Unconjugated(SL0664R) 1:400, overnight at 4癈, followed by conjugation to the secondary antibody(SP-0023) and DAB(SLC0010) stainingHepG2 cell; 4% Paraformaldehyde-fixed; Triton X-100 at room temperature for 20 min; Blocking buffer (normal goat serum, SLC0005) at 37°C for 20 min; Antibody incubation with (HMGB1) polyclonal Antibody, Unconjugated (SL0664R) 1:100, 90 minutes at 37°C; followed by a conjugated Goat Anti-Rabbit IgG antibody at 37°C for 90 minutes, DAPI (blue, C02-04002) was used to stain the cell nuclei.Blank control (blue line): MCF7 (blue). Primary Antibody (green line): Rabbit Anti-HMGB1 antibody (SL0664R) Dilution: 1μg /10^6 cells; Isotype Control Antibody (orange line): Rabbit IgG . Secondary Antibody (white blue line): Goat anti-rabbit IgG-FITCDilution: 1μg /test. ProtocolThe cells were fixed with 70% ethanol (Overnight at 4℃) and then permeabilized with 90% ice-cold methanol for 30 min on ice. Cells stained with Primary Antibody for 30 min at room temperature. The cells were then incubated in 1 X PBS/2%BSA/10% goat serum to block non-specific protein-protein interactions followed by the antibody for 15 min at room temperature. The secondary antibody used for 40 min at room temperature. Acquisition of 20,000 events was performed.Blank control:HL-60. Primary Antibody (green line): Rabbit Anti-HMGB1 antibody (SL0664R) Dilution: 1μg /10^6 cells; Isotype Control Antibody (orange line): Rabbit IgG . Secondary Antibody : Goat anti-rabbit IgG-AF488Dilution: 1μg /test. ProtocolThe cells were fixed with 4% PFA (10min at room temperature)and then permeabilized with 90% ice-cold methanol for 20 min at-20℃. The cells were then incubated in 5%BSA to block non-specific protein-protein interactions for 30 min at room temperature .Cells stained with Primary Antibody for 30 min at room temperature. The secondary antibody used for 40 min at room temperature. Acquisition of 20,000 events was performed.Rat splenocytes stained with Anti- HMGB1 Polyclonal Antibody, A488 Conjugated (SL0664R-A488) at 1:50.The figure annotation: The blue histogram is unstained cells.The Orange histogram is cells stained with Rabbit IgG/FITC (SL0295P-FITC)The green histogram is cells stained with Rabbit Anti-HMGB1/FITC Conjugated antibody (SL0664R-FITC).ControlsPositive control: HepG 2 cellsIsotype control: Cell lines treated with Rabbit IgG/FITC (SL0295P-FITC). instead of the primary antibody to confirm that primary antibody binding is specific. 5μgin 100 μL 1 X PBS containing 0.5% BSA.Positive control: HepG2 cellsConcebtration: 5μg/10^6 cellsIncubation conditions: Avoid light , 30 minutes on the ice.
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Datasheet:


High Mobility Group Box-1 (HMGB1) is a cytokine implicated in the pathogenesis of rheumatoid arthritis (RA) and other inflammatory diseases. The cholinergic anti-inflammatory pathway, a vagus nerve dependent mechanism, inhibits HMGB1 release in experimental disease models

Function:
DNA binding proteins that associates with chromatin and has the ability to bend DNA. Binds preferentially single-stranded DNA. Involved in V(D)J recombination by acting as a cofactor of the RAG complex. Acts by stimulating cleavage and RAG protein binding at the 23 bp spacer of conserved recombination signal sequences (RSS). Heparin-binding protein that has a role in the extension of neurite-type cytoplasmic processes in developing cells.

Subunit:
Component of the RAG complex composed of core components RAG1 and RAG2, and associated component HMGB1 or HMGB2.

Subcellular Location:
Nucleus. Chromosome.

Similarity:
Belongs to the HMGB family.
Contains 2 HMG box DNA-binding domains.

SWISS:
P09429

Gene ID:
3146

Database links:

Entrez Gene: 282691 Cow

Entrez Gene: 3146 Human

Entrez Gene: 100862258 Mouse

Entrez Gene: 15289 Mouse

Entrez Gene: 25459 Rat

Omim: 163905 Human

SwissProt: P10103 Cow

SwissProt: P09429 Human



近来的研究表明称之为高迁移率族蛋白B-1（HMG-B1）的核内结构蛋白在核外表达时是一种有效的早期炎症介质。
高迁移性Ｂ１组蛋白（ＨＭＧＢ１）： 是一种核结合蛋白，在ＤＮＡ重组、修复、复制和基因转录中起作用。ＨＭＧＢ１也是巨噬细胞分泌的一种介质。
此外，高迁移性Ｂ１组蛋白亦被受刺激的巨噬细胞或单核细胞主动分泌。在这一主动分泌过程中，HMGB1首先经乙酰化并由核内转移至溶酶体内，继而在ATP和溶血磷脂胆碱两种分泌信号指导下转移至细胞外。由坏死细胞被动释放的HMGB1和炎症细胞主动分泌的HMGB1存在分子上的差异。胞外的HMGB1可作为细胞因子参与信号传导，因为它既可识别Toll 样受体（TLR）家族的一些成员，又能与识别晚期糖基化终末产物受体（RAGE）相作用。
HMGB1能启动炎症反应，包括产生多种细胞因子、对某些干细胞产生趋化作用、诱导血管粘附分子、削弱肠上皮细胞的功能等。 Picture

Sample: bone (mouse) Lysate at 40 ug
Primary: Anti- HMGB1(SL0664R)at 1/300 dilution
Secondary: IRDye800CW Goat Anti-Rabbit IgG at 1/20000 dilution
Predicted band size: 25 kD
Observed band size: 27kD
Paraformaldehyde-fixed, paraffin embedded (Rat stomach); Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15min; Block endogenous peroxidase by 3% hydrogen peroxide for 20 minutes; Blocking buffer (normal goat serum) at 37°C for 30min; Antibody incubation with (HMGB1) Polyclonal Antibody, Unconjugated (SL0664R) at 1:400 overnight at 4°C, followed by operating according to SP Kit(Rabbit) (sp-0023) instructions and DAB staining.
Tissue/cell: human endometrium carcinoma; 4% Paraformaldehyde-fixed and paraffin-embedded;
Antigen retrieval: citrate buffer ( 0.01M, pH 6.0 ), Boiling bathing for 15min; Block endogenous peroxidase by 3% Hydrogen peroxide for 30min; Blocking buffer (normal goat serum,SLC0005) at 37℃ for 20 min;
Incubation: Anti-HMGB1 Polyclonal Antibody, Unconjugated(SL0664R) 1:200, overnight at 4癈, followed by conjugation to the secondary antibody(SP-0023) and DAB(SLC0010) staining
Tissue/cell: rat liver tissue; 4% Paraformaldehyde-fixed and paraffin-embedded;
Antigen retrieval: citrate buffer ( 0.01M, pH 6.0 ), Boiling bathing for 15min; Block endogenous peroxidase by 3% Hydrogen peroxide for 30min; Blocking buffer (normal goat serum,SLC0005) at 37℃ for 20 min;
Incubation: Anti-HMGB1 Polyclonal Antibody, Unconjugated(SL0664R) 1:400, overnight at 4癈, followed by conjugation to the secondary antibody(SP-0023) and DAB(SLC0010) staining
HepG2 cell; 4% Paraformaldehyde-fixed; Triton X-100 at room temperature for 20 min; Blocking buffer (normal goat serum, SLC0005) at 37°C for 20 min; Antibody incubation with (HMGB1) polyclonal Antibody, Unconjugated (SL0664R) 1:100, 90 minutes at 37°C; followed by a conjugated Goat Anti-Rabbit IgG antibody at 37°C for 90 minutes, DAPI (blue, C02-04002) was used to stain the cell nuclei.
Blank control (blue line): MCF7 (blue).
Primary Antibody (green line): Rabbit Anti-HMGB1 antibody (SL0664R)
Dilution: 1μg /10^6 cells;
Isotype Control Antibody (orange line): Rabbit IgG .
Secondary Antibody (white blue line): Goat anti-rabbit IgG-FITC
Dilution: 1μg /test.
Protocol
The cells were fixed with 70% ethanol (Overnight at 4℃) and then permeabilized with 90% ice-cold methanol for 30 min on ice. Cells stained with Primary Antibody for 30 min at room temperature. The cells were then incubated in 1 X PBS/2%BSA/10% goat serum to block non-specific protein-protein interactions followed by the antibody for 15 min at room temperature. The secondary antibody used for 40 min at room temperature. Acquisition of 20,000 events was performed.
Blank control:HL-60.
Primary Antibody (green line): Rabbit Anti-HMGB1 antibody (SL0664R)
Dilution: 1μg /10^6 cells;
Isotype Control Antibody (orange line): Rabbit IgG .
Secondary Antibody : Goat anti-rabbit IgG-AF488
Dilution: 1μg /test.
Protocol
The cells were fixed with 4% PFA (10min at room temperature)and then permeabilized with 90% ice-cold methanol for 20 min at-20℃. The cells were then incubated in 5%BSA to block non-specific protein-protein interactions for 30 min at room temperature .Cells stained with Primary Antibody for 30 min at room temperature. The secondary antibody used for 40 min at room temperature. Acquisition of 20,000 events was performed.
Rat splenocytes stained with Anti- HMGB1 Polyclonal Antibody, A488 Conjugated (SL0664R-A488) at 1:50.
The figure annotation:
The blue histogram is unstained cells. The Orange histogram is cells stained with Rabbit IgG/FITC (SL0295P-FITC) The green histogram is cells stained with Rabbit Anti-HMGB1/FITC Conjugated antibody (SL0664R-FITC).
Controls
Positive control: HepG 2 cells Isotype control: Cell lines treated with Rabbit IgG/FITC (SL0295P-FITC). instead of the primary antibody to confirm that primary antibody binding is specific. 5μgin 100 μL 1 X PBS containing 0.5% BSA.
Positive control: HepG2 cells
Concebtration: 5μg/10^6 cells
Incubation conditions: Avoid light , 30 minutes on the ice.
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