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Human/Mouse HMGB1/HMG-1 Antibody

Catalog #: AF1690
5 Citations Datasheet / COA / SDS
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AF1690
AF1690-SP
Detection of Human and Mouse HMGB1/HMG‑1 by Western Blot.
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Citations (5)
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Human/Mouse HMGB1/HMG-1 Antibody Summary

Species Reactivity
Human, Mouse
Specificity
Detects human and mouse HMGB1/HMG‑1 in direct ELISAs and Western blots. In direct ELISAs, approximately 15% cross-reactivity with recombinant human (rh) HMGB3 is observed, and less than 1% cross-reactivity with rhHMGA1, rhHMGA2, and rhHMGA1b is observed.  
Source
Polyclonal Chicken IgY
Purification
Antigen Affinity-purified
Immunogen
Mouse myeloma cell line NS0-derived recombinant human HMGB1/HMG‑1
Met1-Glu215
Accession # P09429.3
Formulation
Lyophilized from a 0.2 μm filtered solution in PBS with Trehalose. *Small pack size (SP) is supplied either lyophilized or as a 0.2 µm filtered solution in PBS.
Label
Unconjugated

Applications

Recommended Concentration
Sample
Western Blot
1 µg/mL
See below
Knockout Validated
HMGB1/HMG‑1 is specifically detected in HEK293T human embryonic kidney parental cell line but is not detectable in HMGB1/HMG‑1 knockout HEK293T cell line.
 

Please Note: Optimal dilutions should be determined by each laboratory for each application. General Protocols are available in the Technical Information section on our website.

Scientific Data

Western Blot Detection of Human and Mouse HMGB1/HMG-1 antibody by Western Blot. View Larger

Detection of Human and Mouse HMGB1/HMG‑1 by Western Blot. Western blot shows lysates of HeLa human cervical epithelial carcinoma cell line and mouse lactating tissues. PVDF membrane was probed with 1 µg/mL of Chicken Anti-Human/Mouse HMGB1/HMG‑1 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF1690) followed by HRP-conjugated Anti-Chicken IgY Secondary Antibody. A specific band was detected for HMGB1/HMG‑1 at approximately 25 kDa (as indicated). This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 8.

Knockout Validated Western Blot Shows Human HMGB1/HMG-1 Antibody Specificity by Using Knockout Cell Line. View Larger

Western Blot Shows Human HMGB1/HMG‑1 Specificity by Using Knockout Cell Line. Western blot shows lysates of HEK293T human embryonic kidney parental cell line and HMGB1/HMG-1 knockout HEK293T cell line (KO). PVDF membrane was probed with 1 µg/mL of Chicken Anti-Human/Mouse HMGB1/HMG-1 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF1690) followed by HRP-conjugated Anti-Chicken IgY Secondary Antibody. A specific band was detected for HMGB1/HMG-1 at approximately 30 kDa (as indicated) in the parental HEK293T cell line, but is not detectable in knockout HEK293T cell line. GAPDH (Catalog # AF5718) is shown as a loading control. This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 1.

Reconstitution Calculator

Reconstitution Calculator

The reconstitution calculator allows you to quickly calculate the volume of a reagent to reconstitute your vial. Simply enter the mass of reagent and the target concentration and the calculator will determine the rest.

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Preparation and Storage

Reconstitution
Reconstitute at 0.2 mg/mL in sterile PBS.
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Shipping
The product is shipped at ambient temperature. Upon receipt, store it immediately at the temperature recommended below. *Small pack size (SP) is shipped with polar packs. Upon receipt, store it immediately at -20 to -70 °C
Stability & Storage
Use a manual defrost freezer and avoid repeated freeze-thaw cycles.
  • 12 months from date of receipt, -20 to -70 °C as supplied.
  • 1 month, 2 to 8 °C under sterile conditions after reconstitution.
  • 6 months, -20 to -70 °C under sterile conditions after reconstitution.

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 25-30 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.

References
  1. Lotze, M.T. and K.J. Tracey (2005) Nat. Rev. Immunol. 5:331.
  2. Yang, H. et al. (2005) J. Leukoc. Biol. 78:1.
  3. Dumitriu, I.E. et al. (2005) Trends Immunol. 26:381.
  4. Degryse, B. and M. de Virgilio (2003) FEBS Lett. 553:11.
  5. Wen, L. et al. (1989) Nucleic Acids Res. 17:1197.
  6. Bonaldi, T. et al. (2003) EMBO J. 22:5551.
  7. Muller, S. et al. (2001) EMBO J. 20:4337.
  8. Bustin, M. (1999) Mol. Cell. Biol. 19:5237.
  9. Wang, H. et al. (1999) Science. 285:248.
  10. Dimitriu, I.E. et al. (2005) J. Immunol. 174:7506.
  11. Najima, Y. et al. (2005) J. Biol. Chem. 280:27523.
  12. Gardella, S. et al. (2002) EMBO Rep. 3:995.
Long Name
High Mobility Group Protein 1
Entrez Gene IDs
3146 (Human)
Alternate Names
Amphoterin; high mobility group box 1; High mobility group protein 1; high mobility group protein B1; high-mobility group (nonhistone chromosomal) protein 1; high-mobility group box 1; HMG1; HMG-1; HMG1DKFZp686A04236; HMG3; HMGB1; SBP-1; Sulfoglucuronyl carbohydrate binding protein

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Citations for Human/Mouse HMGB1/HMG-1 Antibody

R&D Systems personnel manually curate a database that contains references using R&D Systems products. The data collected includes not only links to publications in PubMed, but also provides information about sample types, species, and experimental conditions.

5 Citations: Showing 1 - 5
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  1. The C5a/C5aR2 axis promotes renal inflammation and tissue damage
    Authors: T Zhang, KY Wu, N Ma, LL Wei, M Garstka, W Zhou, K Li
    JCI Insight, 2020-04-09;5(7):.
    Species: Mouse
    Sample Types: Tissue Lysates
    Applications: Western Blot
  2. Cytosolic HMGB1 controls the cellular autophagy/apoptosis checkpoint during inflammation.
    Authors: Zhu X, Messer J, Wang Y, Lin F, Cham C, Chang J, Billiar T, Lotze M, Boone D, Chang E
    J Clin Invest, 2015-02-02;125(3):1098-110.
    Species: Mouse
    Sample Types: Cell Lysates
    Applications: Western Blot
  3. Cancer usurps skeletal muscle as an energy repository.
    Authors: Luo, Yi, Yoneda, Junya, Ohmori, Hitoshi, Sasaki, Takamits, Shimbo, Kazutaka, Eto, Sachise, Kato, Yumiko, Miyano, Hiroshi, Kobayashi, Tsuyoshi, Sasahira, Tomonori, Chihara, Yoshitom, Kuniyasu, Hiroki
    Cancer Res, 2013-11-06;74(1):330-40.
    Species: Mouse
    Sample Types: In Vivo
    Applications: Neutralization
  4. 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, 2012-08-10;7(8):e42748.
    Species: Human
    Sample Types: Sputum
    Applications: ELISA Development
  5. Berberine inhibits the ischemia-reperfusion injury induced inflammatory response and apoptosis of myocardial cells through the phosphoinositide 3-kinase/RAC-? serine/threonine-protein kinase and nuclear factor-?B signaling pathways
    Authors: L Wang, H Ma, Y Xue, H Shi, T Ma, X Cui
    Exp Ther Med, 2017-11-27;15(2):1225-1232.

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