Anti-HMGB1 (HMG1) (N-terminal) antibody produced in rabbit
SIGMA/H9664 - affinity isolated antibody, buffered aqueous solution
Synonym: Anti-Amphoterin; Anti-High mobility group box 1; Anti-High mobility group protein 1
MDL Number: MFCD02263062
Product Type: Chemical
Enhanced Validation-By Independent Antibodies Immunofluoresence. PC12 cells were fixed and permeabilized with 4% paraformaldehyde followed by 0.5% Triton X-100. Fixed cells were stained with 1 μg/mL Anti-HMGB1 (HMG1) (C-terminal) antibody produced in Rabbit (Cat. No. H9539) (A). The antibody developed using Goat Anti-Rabbit IgG, Cy3 Conjugate antibody, and 1 μg/mL Anti-HMGB1 (HMG1) (N-terminal) antibody produced in Rabbit (Cat. No. H9664) (B). The antibody was developed using Goat Anti-Rabbit IgG, Cy3 Conjugate antibody. Results Two Anti HMGB1 (HMG1) antibodies, H9539 (A) and H9664 (B) target different regions of HMGB1 (HMG1) show similar staining profiles between the two antibodies, demonstrating Independent Antibody Verification.
Immunofluorescence Anti-HMGB1 : Cat. No. H9664: Immunofluorescence of HUVEC cells using Anti-HMGB1 (red) at a 1:50 dilution, taken at 40× magnification and nuclear staining with Hoescht 33342 (blue). Yale HTCB IF procedure used.
Immunofluorescence Nuclear localization of HMGB1 in PC12 cells. A. Cells were stained with Rabbit Anti HMGB1 (HMG1) (N-Terminal) (Cat. No. H9664) at 1 μg/mL. B. DAPI staining. C. Merged staining of HMGB1 and DAPI indicating nuclear staining of HMGB1.
Immunofluorescence Graph Anti-HMGB1 : Cat. No. H9664: Intensity analysis of Anti-HMGB1 staining. Cytoplasmic and nuclear intensity values were obtained at 8 different antibody dilutions (1:50 to 1:6,400) as compared with the negative control. Yale HTCB IF procedure used.
Immunoblotting Lysates of various tissues and cell lines were separated by SDS-PAGE, blotted with 2 mg/mL rabbit anti HMGB1 (N-Terminal) (Cat. No. H9664) and developed with Goat Anti-Rabbit IgG, Peroxidase conjugate (Cat. No. A0545) using a chemiluminescence substrate. Lanes 1. mouse kidney, 2. NIH3T3, 3. Rat1, 4. RINSH, 5. PC12, 6. K562; 7. HEK-293T, 8. HeLa nuclear, 9. HeLa
Immunoprecipitation Lane 1.10 mg of Rabbit Anti-HMGB1 (Cat. No. H9664) was used to immunoprecipitate human HMGB1 from HEK-293T cell lysate. Lane 2. Negative control – without cell lysate. Lane 3. Negative control – without IP antibody. Detection antibody: Rabbit anti HMGB1 (HMG1) (C-Terminal) (Cat. No. H9539)
| antibody form | affinity isolated antibody |
| antibody product type | primary antibodies |
| biological source | rabbit |
| clone | polyclonal |
| conjugate | unconjugated |
| enhanced validation | independent Learn more about Antibody Enhanced Validation  |
| form | buffered aqueous solution |
| mol wt | antigen 25 kDa |
| Quality Level | 200 |
| shipped in | dry ice |
| species reactivity | human, rat, mouse |
| storage temp. | −20°C |
| target post-translational modification | unmodified |
| technique(s) | immunoprecipitation (IP): 10 μg using HEK-293T cell lysates |
| indirect immunofluorescence: 1-2 μg/mL using paraformaldehyde-Triton fixed PC12 cultured cells. | |
| indirect immunofluorescence: suitable | |
| western blot: 1-2 μg/mL using 3T3 cell lysates | |
| UniProt accession no. | P09429  |
| Application: | Anti-HMGB1 (HMG1) (N-terminal) antibody is suitable for use in chemiluminescent immunoblot (using mouse heart homogenates) . The antibody can also be used in immunoprecipitation (10 μg using HEK-293T cell lysates), indirect immunofluorescence (1-2 μg/mL using paraformaldehyde-Triton fixed PC12 cultured cells), and western blot (1-2 μg/mL using 3T3 cell lysates). |
| Application: | Applications in which this antibody has been used successfully, and the associated peer-reviewed papers, are given below. Western Blotting (1 paper) |
| Application: | Yale Center for High Throughput Cell Biology IF-tested antibodies. Each antibody is tested by immunofluorescence against HUVEC cells using the Yale HTCB IF protocol. To learn more about us and Yale Center for High Throughput Cell Biology partnership, visit sigma.com/htcb-if . |
| Biochem/physiol Actions: | High Mobility Group B (HMGB-1 and 2) participate in the regulation of chromatin structure as well as being involved in transcription regulation, DNA repair, recombination, differentiation and extracellular signaling. It shows a defect in transcriptional enhancement of the glucocorticoid receptor, that indicates the important role for HMGB1 in proper transcriptional control by specific transcription factors. Increased expression of HMGB1 is observed in cancer cells. |
| Disclaimer: | Unless otherwise stated in our catalog or other company documentation accompanying the product(s), our products are intended for research use only and are not to be used for any other purpose, which includes but is not limited to, unauthorized commercial uses, in vitro diagnostic uses, ex vivo or in vivo therapeutic uses or any type of consumption or application to humans or animals. |
| General description: | Anti-HMGB1 (HMG1) (N-terminal) is produced in rabbit using as immunogen a synthetic peptide corresponding to human HMGB1 conjugated to KLH. High Mobility Group B (HMGB) protein family includes HMGB1, HMGB2 and HMGB3, that are highly conserved and indistinguishable in their biochemical properties. HMGB1 is a 25 kDa protein of 215 amino acids and consists of two homologous HMG-boxes rich in basic amino acids, and an acidic tail at the carboxy-terminus. |
| General description: | HMGB proteins belong to the High Mobility Group (HMG) family of proteins that contain the HMG-box for binding and changing DNA structures . |
| Immunogen: | synthetic peptide corresponding to amino acids 2-17 of human HMGB1, conjugated to KLH. The corresponding sequence is conserved in mouse and rat. |
| Physical form: | Solution in 0.01 M phosphate buffered saline, pH 7.4, containing 15 mM sodium azide. |
| RIDADR | NONH for all modes of transport |
| Storage Temp. | −20°C |
| UNSPSC | 12352203 |