PRODUCT CODE: ER0913

HMGB1 Rabbit Polyclonal Antibody (ER0913)

Applications

  • WB

  • ICC

  • IHC-P

  • FC

REACTIVITY

  • Human

  • Mouse

  • Rat

All lanes: Western blot analysis of HMGB1 with anti-HMGB1 antibody (ER0913) at 1:500 dilution.<br />
Lane 1: Wild-type Raw264.7 whole cell lysate.<br />
Lane 2: HMGB1 knockdown Raw264.7 whole cell lysate.<br />
<br />
ER0913 was shown to specifically react with HMGB1 in Wild-type Raw264.7 cells. Weakened band was observed when HMGB1 knockdown samples were tested. Wild-type and HMGB1 knockdown samples were subjected to SDS-PAGE. Proteins were transferred to a PVDF membrane and blocked with 5% NFDM in TBST for 1 hour at room temperature. The primary Anti-HMGB1 antibody (ER0913, 1/500) and Anti-HSP90 antibody (ET1605-56, 1/10,000) were used in 5% BSA at room temperature for 2 hours. Goat Anti-Rabbit IgG H&L (HRP) Secondary Antibody (HA1001) at 1:200,000 dilution was used for 1 hour at room temperature.
  • All lanes: Western blot analysis of HMGB1 with anti-HMGB1 antibody (ER0913) at 1:500 dilution.<br />
Lane 1: Wild-type Raw264.7 whole cell lysate.<br />
Lane 2: HMGB1 knockdown Raw264.7 whole cell lysate.<br />
<br />
ER0913 was shown to specifically react with HMGB1 in Wild-type Raw264.7 cells. Weakened band was observed when HMGB1 knockdown samples were tested. Wild-type and HMGB1 knockdown samples were subjected to SDS-PAGE. Proteins were transferred to a PVDF membrane and blocked with 5% NFDM in TBST for 1 hour at room temperature. The primary Anti-HMGB1 antibody (ER0913, 1/500) and Anti-HSP90 antibody (ET1605-56, 1/10,000) were used in 5% BSA at room temperature for 2 hours. Goat Anti-Rabbit IgG H&L (HRP) Secondary Antibody (HA1001) at 1:200,000 dilution was used for 1 hour at room temperature.
  • Western blot analysis of HMGB1 on different cell lysates using anti- HMGB1 antibody at 1/500 dilution.<br />
Positive control:    <br />
Lane 1: HepG2    <br />
Lane 2: MCF-7  <br />
Lane 3: F9        <br />
Lane 4: A549
  • ICC staining HMGB1 in Hela cells (green). Cells were fixed in paraformaldehyde, permeabilised with 0.25% Triton X100/PBS.
  • ICC staining HMGB1 in NIH/3T3 cells (green). Cells were fixed in paraformaldehyde, permeabilised with 0.25% Triton X100/PBS.
  • Immunohistochemical analysis of paraffin-embedded mouse kidney tissue using anti- HMGB1 antibody. Counter stained with hematoxylin.
  • Immunohistochemical analysis of paraffin-embedded mouse stomach tissue using anti- HMGB1 antibody. Counter stained with hematoxylin.
  • Immunohistochemical analysis of paraffin-embedded mouse brain tissue using anti- HMGB1 antibody. Counter stained with hematoxylin.
  • Immunohistochemical analysis of paraffin-embedded human stomach carcinoma tissue using anti- HMGB1 antibody. Counter stained with hematoxylin.
  • Immunohistochemical analysis of paraffin-embedded human tonsil tissue using anti- HMGB1 antibody. Counter stained with hematoxylin.
  • Flow cytometric analysis of HMGB1 cells with HMGB1 antibody at 1/50 dilution (blue) compared with an unlabelled control (cells without incubation with primary antibody; red). Goat anti rabbit IgG (FITC) was used as the secondary antibody.
All lanes: Western blot analysis of HMGB1 with anti-HMGB1 antibody (ER0913) at 1:500 dilution.
Lane 1: Wild-type Raw264.7 whole cell lysate.
Lane 2: HMGB1 knockdown Raw264.7 whole cell lysate.

ER0913 was shown to specifically react with HMGB1 in Wild-type Raw264.7 cells. Weakened band was observed when HMGB1 knockdown samples were tested. Wild-type and HMGB1 knockdown samples were subjected to SDS-PAGE. Proteins were transferred to a PVDF membrane and blocked with 5% NFDM in TBST for 1 hour at room temperature. The primary Anti-HMGB1 antibody (ER0913, 1/500) and Anti-HSP90 antibody (ET1605-56, 1/10,000) were used in 5% BSA at room temperature for 2 hours. Goat Anti-Rabbit IgG H&L (HRP) Secondary Antibody (HA1001) at 1:200,000 dilution was used for 1 hour at room temperature.

Applications

  • WB

  • ICC

  • IHC-P

  • FC

REACTIVITY

  • Human

  • Mouse

  • Rat

SPECIFICATIONS

Product Type

Rabbit polyclonal primary

Product Name

HMGB1 Rabbit Polyclonal Antibody (ER0913)

Immunogen

Synthetic peptide within human hmgb1 aa 71-126.

Host

Rabbit

Positive Control

HepG2, MCF-7, F9, A549, PC12, Hela, NIH/3T3, RAW264.7, mouse kidney tissue Mouse stomach tissue, mouse brain tissue, human stomach carcinoma tissue, human tonsil tissue

Conjugation

Unconjugated

Clonality

Polyclonal

PROPERTIES

Form

Liquid

Storage Condition

Store at +4C after thawing. Aliquot store at -20C or -80C. Avoid repeated freeze / thaw cycles.

Storage Buffer

1*PBS (pH7.4), 0.2% BSA, 40% Glycerol. Preservative: 0.05% Sodium Azide.

Concentration

1 ug/ul

PURIFICATION

Peptide affinity purified

MOLECULAR WEIGHT

25 kDa

Isotype

IgG

APPLICATION DILUTION

  • WB

  • 1:500-1:1,000

  • ICC

  • 1:200

  • IHC-P

  • 1:200

  • FC

  • 1:50-1:100

TARGET

UNIPROT #

PROTEIN NAME

HMGB1

SYNONYMS

Amphoterin antibody; Chromosomal protein, nonhistone, HMG1 antibody; DKFZp686A04236 antibody; High mobility group 1 antibody; High mobility group box 1 antibody; High mobility group protein 1 antibody; High mobility group protein B1 antibody; high-mobility group (nonhistone chromosomal) protein 1 antibody; HMG-1 antibody; HMG1 antibody; HMG3 antibody; HMGB 1 antibody; HMGB1 antibody; HMGB1_HUMAN antibody; NONHISTONE CHROMOSOMAL PROTEIN HMG1 antibody; SBP 1 antibody; Sulfoglucuronyl carbohydrate binding protein antibody

SEQUENCE SIMILARITIES

Belongs to the HMGB family.

TISSUE SPECIFICITY

Ubiquituous. Expressed in platelets.

POST-TRANSLATIONAL MODIFICATION

Phosphorylated at serine residues. Phosphorylation in both NLS regions is required for cytoplasmic translocation followed by secretion.; Acetylated on multiple sites upon stimulation with LPS. Acetylation on lysine residues in the nuclear localization signals (NLS 1 and NLS 2) leads to cytoplasmic localization and subsequent secretion (By similarity). Acetylation on Lys-3 results in preferential binding to DNA ends and impairs DNA bending activity (By similarity).; Reduction/oxidation of cysteine residues Cys-23, Cys-45 and Cys-106 and a possible intramolecular disulfide bond involving Cys-23 and Cys-45 give rise to different redox forms with specific functional activities in various cellular compartments: 1- fully reduced HMGB1 (HMGB1C23hC45hC106h), 2- disulfide HMGB1 (HMGB1C23-C45C106h) and 3- sulfonyl HMGB1 (HMGB1C23soC45soC106so).; Poly-ADP-ribosylated by PARP1 when secreted following stimulation with LPS (By similarity).; In vitro cleavage by CASP1 is liberating a HMG box 1-containing peptide which may mediate immunogenic activity; the peptide antagonizes apoptosis-induced immune tolerance. Can be proteolytically cleaved by a thrombin:thrombomodulin complex; reduces binding to heparin and proinflammatory activities (By similarity).

SUBCELLULAR LOCATION

Nucleus

FUNCTION

Like the histones, HMGB1, also known as high-mobility group protein 1 (HMG-1) is among the most important chromatin proteins. In the nucleus HMGB1 interacts with nucleosomes, transcription factors, and histones. This nuclear protein organizes the DNA and regulates transcription. After binding, HMGB1 bends DNA, which facilitates the binding of other proteins. HMGB1 is secreted by immune cells through leaderless secretory pathway. Activated macrophages and monocytes secrete HMGB1 as a cytokine mediator of Inflammation. In recent research, HMGB1 has been reported as a novel biomarker for human ovarian cancer.