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:
SL20633R
Species Reactivity:
Human,Mouse,Rat,(predicted: Dog,Pig,Cow,Horse,)
Immunogen:
KLH conjugated synthetic peptide derived from mouse HMGB1:61-150/215
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/TestIF=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:MCF-7 Cell (human) Lysate at 40 ugPrimary: Anti- HMGB1 (SL20633R)at 1/300 dilutionSecondary: IRDye800CW Goat Anti-Rabbit IgG at 1/20000 dilutionPredicted band size: 25kDObserved band size: 27 kD 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 (SL20633R) at 1:500 overnight at 4°C, followed by a conjugated secondary (sp-0023) for 20 minutes and DAB staining. Paraformaldehyde-fixed, paraffin embedded (Rat brain); 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 (SL20633R) at 1:500 overnight at 4°C, followed by a conjugated secondary (sp-0023) for 20 minutes and DAB staining. Paraformaldehyde-fixed, paraffin embedded (Mouse brain); 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 (SL20633R) at 1:500 overnight at 4°C, followed by a conjugated secondary (sp-0023) for 20 minutes and DAB staining. Paraformaldehyde-fixed, paraffin embedded (Rat liver); 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 (SL20633R) at 1:500 overnight at 4°C, followed by a conjugated secondary (sp-0023) for 20 minutes and DAB staining. This image was generously provided by Juanli Fu, at Southwest University in Chong Qing, China. 4% Paraformaldehyde fixed PC12 cells stained with Rabbit Anti- HMGB1 Polyclonal Antibody (SL20633R) at 1:300 for 3 hours at 4°C, followed by Rhodamine-conjugated secondary antibody for an additional hour.Blank control (blue line): MCF7 (fixed with 80% ethanol (Overnight at 4℃) and then permeabilized with 90% ice-cold methanol for 30 min on ice). Primary Antibody (green line): Rabbit Anti-HMGB1 antibody (SL20633R),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. Blank control:HL-60. Primary Antibody (green line): Rabbit Anti-HMGB1 antibody (SL20633R) 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.
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Product PDFs
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:
100862258

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:
MCF-7 Cell (human) Lysate at 40 ug
Primary: Anti- HMGB1 (SL20633R)at 1/300 dilution
Secondary: IRDye800CW Goat Anti-Rabbit IgG at 1/20000 dilution
Predicted band size: 25kD
Observed band size: 27 kD
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 (SL20633R) at 1:500 overnight at 4°C, followed by a conjugated secondary (sp-0023) for 20 minutes and DAB staining.
Paraformaldehyde-fixed, paraffin embedded (Rat brain); 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 (SL20633R) at 1:500 overnight at 4°C, followed by a conjugated secondary (sp-0023) for 20 minutes and DAB staining.
Paraformaldehyde-fixed, paraffin embedded (Mouse brain); 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 (SL20633R) at 1:500 overnight at 4°C, followed by a conjugated secondary (sp-0023) for 20 minutes and DAB staining.
Paraformaldehyde-fixed, paraffin embedded (Rat liver); 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 (SL20633R) at 1:500 overnight at 4°C, followed by a conjugated secondary (sp-0023) for 20 minutes and DAB staining.
This image was generously provided by Juanli Fu, at Southwest University in Chong Qing, China. 4% Paraformaldehyde fixed PC12 cells stained with Rabbit Anti- HMGB1 Polyclonal Antibody (SL20633R) at 1:300 for 3 hours at 4°C, followed by Rhodamine-conjugated secondary antibody for an additional hour.
Blank control (blue line): MCF7 (fixed with 80% ethanol (Overnight at 4℃) and then permeabilized with 90% ice-cold methanol for 30 min on ice).
Primary Antibody (green line): Rabbit Anti-HMGB1 antibody (SL20633R),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.
Blank control:HL-60.
Primary Antibody (green line): Rabbit Anti-HMGB1 antibody (SL20633R)
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.
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