抗Human HMGB1抗体(Anti-Human HMGB1 antibody)

• 価格
• Product Details
• Applications and Data
• Related Product & Information
• Citations

価格

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Product Details

Species Reactivity	Human
Label	Unconjugated
Immunogen	E. coli-derived recombinant human HMGB1/HMG-1Gly2-Glu215 (predicted)Accession # P09429
Source	Monoclonal Mouse IgG2B Clone # 115603
Purification	Protein A or G purified from hybridoma culture supernatant
Specificity	Detects human HMGB1/HMG-1 in direct ELISAs and Western blots.

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Applications and Data

	Recommended Concentration	Sample
Western Blot	0.05 µg/mL	See below
Chromatin Immunoprecipitation (ChIP)	5 µg/5 x 106 cells	See below
CyTOF-ready	Ready to be labeled using established conjugation methods. No BSA or other carrier proteins that could interfere with conjugation.
Immunocytochemistry	8-25 µg/mL	See below
Intracellular Staining by Flow Cytometry	2.5 µg/106 cells	See below

Western Blot
	Detection of Human HMGB1/HMG‑1 by Western Blot. Western blot shows lysates of HepG2 human hepatocellular carcinoma cell line, Jurkat human acute T cell leukemia cell line, and HEK293 human embryonic kidney cell line. PVDF membrane was probed with 0.05 µg/mL of Mouse Anti-Human HMGB1/HMG‑1 Monoclonal Antibody (Catalog # MAB1690) followed by HRP-conjugated Anti-Mouse IgG Secondary Antibody (Catalog # HAF018). A specific band was detected for HMGB1/HMG‑1 at approximately 26 kDa (as indicated). This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 1.
Chromatin Immunoprecipitation (ChIP)
	Detection of HMGB1/HMG‑1-regulated Genes by Chromatin Immunoprecipitation. HepG2 human hepatocellular carcinoma cell line was fixed using formaldehyde, resuspended in lysis buffer, and sonicated to shear chromatin. HMGB1/HMG‑1/DNA complexes were immunoprecipitated using 5 μg Mouse Anti-Human HMGB1/HMG‑1 PE-conjugated Monoclonal Antibody (Catalog # MAB1690) or control antibody (Catalog # MAB004) for 15 minutes in an ultrasonic bath, followed by Biotinylated Anti-Mouse IgG Secondary Antibody (Catalog # BAF007). Immunocomplexes were captured using 50 μL of MagCellect Streptavidin Ferrofluid (Catalog # MAG999) and DNA was purified using chelating resin solution. The AFP promoter was detected by standard PCR.
Intracellular Staining by Flow Cytometry
	Detection of HMGB1 in HCT‑116 Human Cell Line by Flow Cytometry. HCT‑116 human colorectal carcinoma cell line was stained with Mouse Anti-Human HMGB1/HMG-1 Monoclonal Antibody (Catalog # MAB1690, filled histogram) or isotype control antibody (Catalog # MAB0041, open histogram), followed by Phycoerythrin-conjugated Anti-Mouse IgG F(ab')2 Secondary Antibody (Catalog # F0102B). To facilitate intracellular staining, cells were fixed with Flow Cytometry Fixation Buffer (Catalog # FC004) and permeabilized with Flow Cytometry Permeabilization/Wash Buffer I (Catalog # FC005). View our protocol for Staining Intracellular Molecules.
Immunocytochemistry
	HMGB1/HMG‑1 in CCD‑18Co Human Cell Line. HMGB1/HMG‑1 was detected in immersion fixed CCD‑18Co human colonic fibroblast cell line using Mouse Anti-Human HMGB1/HMG‑1 Monoclonal Antibody (Catalog # MAB1690) at 10 µg/mL for 3 hours at room temperature. Cells were stained using the NorthernLights™ 557-conjugated Anti-Mouse IgG Secondary Antibody (red, upper panel; Catalog # NL007) and counterstained with DAPI (blue, lower panel). Specific staining was localized to nuclei. View our protocol for Fluorescent ICC Staining of Cells on Coverslips.

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Related Product & Information

Background

HMGB1/HMG-1

background_content

Background: HMGB1/HMG-1 Human High-mobility group box 1 protein (HMGB1), previously known as HMG-1 or amphoterin, is a member of the high mobility group box family of non-histone chromosomal proteins (1‑3). Human HMGB1 is expressed as a 30 kDa, 215 amino acid (aa) single chain polypeptide containing three domains: two N-terminal globular, 70 aa positively charged DNA-binding domains (HMG boxes A and B), and a negatively charged 30 aa C-terminal region that contains only Asp and Glu (4, 5). Residues 27‑43 and 178‑184 contain a NLS. Posttranslational modifications of the molecule have been reported, with acetylation occurring on as many as 17 lysine residues (6). HMGB1 is expressed at high levels in almost all cells (2, 4). It was originally discovered as a nuclear protein that could bend DNA. Such bending stabilizes nucleosome formation and regulates the expression of select genes upon recruitment by DNA binding proteins (1, 7, 8). It is now known that HMGB1 can also act extracellularly, both as an inflammatory mediator that promotes monocyte migration and cytokine secretion, and as a mediator of T cell-dendritic cell interaction (1, 4, 7, 9, 10). The cytokine activity of HBMG1 is restricted to the HMG B box, (3) while the A box is associated with the helix-loop-helix domain of transcription factors (11). HMBG1 is released in response to cell death and as a secretion product. Although HMBG-1 does not possess a classic signal sequence, it appears to be secreted as an acetylated form via secretory endolysosome exocytosis (6, 12). Once secreted, HMGB1 transduces cellular signals through its high affinity receptor, RAGE and, possibly, TLR2 and TLR4 (1, 3, 4). Human HMGB1 is 100% aa identical to canine HMGB1 and 99% aa identical to mouse, rat, bovine and porcine HMGB1, respectively.

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Citations

1. Anti-high mobility group box-1 (HMGB1) antibody attenuates delayed cerebral vasospasm and brain injury after subarachnoid hemorrhage in rats
   Sci Rep, 2016;6(0):37755.
   Species: Rat
   Sample Type: Whole Tissue
   Application: IHC Paraffin-embedded


2. Expression of RAGE and HMGB1 in thymic epithelial tumors, thymic hyperplasia and regular thymic morphology.
   Authors: Moser B, Janik S, Schiefer A, Mullauer L, Bekos C, Scharrer A, Mildner M, Renyi-Vamos F, Klepetko W, Ankersmit H
   PLoS ONE, 2014;9(4):e94118.
   Species: Human
   Sample Type: Whole Tissue
   Application: IHC Paraffin-embedded


3. A dual-role of Gu-4 in suppressing HMGB1 secretion and blocking HMGB1 pro-inflammatory activity during inflammation.
   Authors: Zhou H, Ji X, Wu Y, Xuan J, Qi Z, Shen L, Lan L, Li Q, Yin Z, Li Z, Zhao Z
   PLoS ONE, 2014;9(3):e89634.
   Species: Mouse
   Sample Type: Whole Cells
   Application: ICC


4. Local and systemic RAGE axis changes in pulmonary hypertension: CTEPH and iPAH.
   Authors: Moser B, Megerle A, Bekos C, Janik S, Szerafin T, Birner P, Schiefer A, Mildner M, Lang I, Skoro-Sajer N, Sadushi-Kolici R, Taghavi S, Klepetko W, Ankersmit H
   PLoS ONE, 2014;9(9):e106440.
   Species: Human
   Sample Type: Whole Tissue
   Application: IHC


5. IL-1alpha released from damaged epithelial cells is sufficient and essential to trigger inflammatory responses in human lung fibroblasts.
   Authors: Suwara M, Green N, Borthwick L, Mann J, Mayer-Barber K, Barron L, Corris P, Farrow S, Wynn T, Fisher A, Mann D
   Mucosal Immunol, 2014;7(3):684-93.
   Species: Human
   Sample Type: Cell Culture Supernates
   Application: ELISA Development


6. Measles virus causes immunogenic cell death in human melanoma.
   Authors: Donnelly, O G, Errington-Mais, F, Steele, L, Hadac, E, Jennings, V, Scott, K, Peach, H, Phillips, R M, Bond, J, Pandha, H, Harrington, K, Vile, R, Russell, S, Selby, P, Melcher, A A
   Gene Ther, 2013;20(1):7-15.
   Species: Human
   Sample Type: Cell Culture Supernates
   Application: WB


7. Sputum biomarkers and the prediction of clinical outcomes in patients with cystic fibrosis.
   Authors: Liou T, Adler F, Keogh R, Li Y, Jensen J, Walsh W, Packer K, Clark T, Carveth H, Chen J, Rogers S, Lane C, Moore J, Sturrock A, Paine R, Cox D, Hoidal J
   PLoS ONE, 2012;7(8):e42748.
   Species: Human
   Sample Type: Sputum
   Application: ELISA Development


8. Human Papillomavirus Type 8 Interferes with a Novel C/EBPβ-Mediated Mechanism of Keratinocyte CCL20 Chemokine Expression and Langerhans Cell Migration.
   Authors: Sperling T, Oldak M, Walch-Ruckheim B, Wickenhauser C, Doorbar J, Pfister H, Malejczyk M, Majewski S, Keates AC, Smola S
   PLoS Pathog., 2012;8(7):e1002833.
   Species: Human
   Sample Type: Cell Lysates
   Application: WB


9. Fecal HMGB1 is a novel marker of intestinal mucosal inflammation in pediatric inflammatory bowel disease.
   Authors: Vitali R, Stronati L, Negroni A
   Am. J. Gastroenterol., 2011;106(11):2029-40.
   Species: Human
   Sample Type: Feces
   Application: WB


10. Ethyl pyruvate decreases HMGB1 release and ameliorates murine colitis.
    Authors: Dave SH, Tilstra JS, Matsuoka K, Li F, DeMarco RA, Beer-Stolz D, Sepulveda AR, Fink MP, Lotze MT, Plevy SE
    J. Leukoc. Biol., 2009;86(3):633-43.
    Species: Mouse
    Sample Type: Feces
    Application: ELISA Development

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