Anti-HMGB1 antibody (ab77302)
Key features and details
- Mouse monoclonal to HMGB1
- Suitable for: WB, IHC-P, ICC/IF, Flow Cyt
- Reacts with: Human
- Isotype: IgG1
Overview
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Product name
Anti-HMGB1 antibody
See all HMGB1 primary antibodies -
Description
Mouse monoclonal to HMGB1 -
Host species
Mouse -
Tested Applications & Species
See all applications and species dataApplication Species Flow Cyt HumanICC/IF HumanIHC-P HumanWB Human
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Immunogen
Recombinant full length protein (GST-tag) corresponding to Human HMGB1 aa 1-215. AAH03378
Sequence:MGKGDPKKPRGKMSSYAFFVQTCREEHKKKHPDASVNFSEFSKKCSERWK TMSAKEKGKFEDMAKADKARYEREMKTYIPPKGETKKKFKDPNAPKRPPS AFFLFCSEYRPKIKGEHPGLSIGDVAKKLGEMWNNTAADDKQPYEKKAAK LKEKYEKDIAAYRAKGKPDAAKKGVVKAEKSKKKKEEEEDEEDEEDEEEE EDEEDEDEEEDDDDE
Database link: P09429 -
Positive control
- WB: HMGB1-transfected HEK-293T cell lysate. IHC-P: Human stomach tissue. ICC/IF: HeLa cells. Flow Cytometry: HeLa cells.
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General notes
This product was changed from ascites to tissue culture supernatant on 15 May 2019. Please note that the dilutions may need to be adjusted accordingly. If you have any questions, please do not hesitate to contact our scientific support team.
Properties
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Form
Liquid -
Storage instructions
Shipped at 4°C. Upon delivery aliquot and store at -20°C or -80°C. Avoid repeated freeze / thaw cycles. -
Storage buffer
pH: 7.40
Constituent: PBS -
Concentration information loading... -
Purity
Tissue culture supernatant -
Purification notes
Purified from TCS. -
Clonality
Monoclonal -
Isotype
IgG1 -
Light chain type
kappa -
Research areas
Images
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Western blot - Anti-HMGB1 antibody (ab77302)All lanes : Anti-HMGB1 antibody (ab77302) at 5 µg/ml
Lane 1 : HMGB1-transfected HEK-293T (Human epithelial cell line from embryonic kidney transformed with large T antigen) cell lysate
Lane 2 : Non-transfected HEK-293T (Human epithelial cell line from embryonic kidney transformed with large T antigen) cell lysate
Lysates/proteins at 25 µg per lane.
Secondary
All lanes : Goat Anti-Mouse IgG (H&L)-HRP Conjugate at 1/2500 dilution
Predicted band size: 25 kDa
Observed band size: 29 kDa why is the actual band size different from the predicted?This image was generated using the ascites version of the product.
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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-HMGB1 antibody (ab77302)Formalin-fixed, paraffin-embedded human stomach tissue stained for HMGB1 with ab77302 at 3 µg/ml in immunohistochemical analysis.
This image was generated using the ascites version of the product.
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Immunocytochemistry/ Immunofluorescence - Anti-HMGB1 antibody (ab77302)HeLa (human epithelial cell line from cervix adenocarcinoma) cells stained for HMGB1 (green) using ab77302 at 10 µg/ml in ICC/IF.
This image was generated using the ascites version of the product.
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Flow Cytometry - Anti-HMGB1 antibody (ab77302)Overlay histogram showing HeLa (Human epithelial cell line from cervix adenocarcinoma) cells stained with ab77302 (red line). The cells were fixed with methanol (5 minutes) and then permeabilized with 0.1% PBS-Tween for 20 minutes. The cells were then incubated in 1x PBS / 10% normal goat serum / 0.3M glycine to block non-specific protein-protein interactions followed by the antibody ab77302, 2 µg/1x106 cells for 30 minutes at 22°C. The secondary antibody used was DyLight® 488 goat anti-mouse IgG (H+L) ab96879 at 1/500 dilution for 30 minutes at 22°C. Isotype control antibody (black line) was mouse mouse IgG1 [ICIGG1] (ab91353 2 µg/1x106 cells) used under the same conditions. Acquisition of >5,000 events was performed. This antibody gave a positive signal in HeLa cells fixed with 4% paraformaldehyde (10 minutes)/permeabilized with 0.1% PBS-Tween 20 used under the same conditions.
This image was generated using the ascites version of the product.
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Immunocytochemistry/ Immunofluorescence - Anti-HMGB1 antibody (ab77302) Image from Mitroulis I et al., PLoS ONE 6(12): e29318. Fig 6.; doi: 10.1371/journal.pone.0029318. Epub 2011 Dec 16.Immunofluorescence analysis of human peripheral polymorphonuclear cells, staining HMGB1 with ab77302 (green).
Cells were fixed in 4% paraformaldehyde and blocked with 5% rabbit serum. Cells were incubated with primary antibody and an AlexaFluor®488-conjugated anti-mouse IgG was used as the secondary antibody. Nuclei were stained with DAPI (blue).This image was generated using the ascites version of the product.
