Anti-HMGB1 antibody - ChIP Grade

• Supplier: Abcam
• Catalog: ab18256
• Prices: $404.00
• Sizes: 100 µg
• Host: Rabbit
• Clonality: Polyclonal
• Isotype: IgG
• Applications: IHC-F ICC/IF ICC/IF IHC-P ChIP IHC-F ICC/IF ELISA IP WB
• Species Reactivities: Mouse, Rat, Human, Rabbit, Bovine
• Antigen: Synthetic peptide conjugated to KLH derived from within residues 150 to the C-terminus of Human HMGB1
• Blocking Peptide: Human HMGB1 peptide
• Description: Rabbit Polyclonal
• Gene: HMGB1
• Conjugate: Unconjugated

Product images

Image courtesy of Human Protein AtlasParaffin embedded sections of human liver were incubated with ab18256 (1/1000 dilution) for 30 minutes at room temperature. Heat induced antigen retrieval was performed in citrate buffer pH 6.ab18256 was tested in a tissue microarray (TMA) containing a wide range of normal and cancer tissues as well as a cell microarray consisting of a range of commonly used, well characterised human cell lines. Further images can be found www.proteinatlas.org

All lanes : Anti-HMGB1 antibody - ChIP Grade (ab18256) at 1/1000 dilutionLane 1 : Rat brain whole tissue lysate - infused with asf for 1 weekLane 2 : Rat brain whole tissue lysate - infused with LPS for 1 weekLane 3 : Rat brain whole tissue lysate - infused with acsf for 8 weeksLane 4 : Rat brain whole tissue lysate - infused with LPS for 8 weeksLane 5 : Rat brain whole tissue lysate - infused with LPS for 4 weeksLane 6 : Rat brain whole tissue lysate -infused with LPS for 4 weeks, after 2 weeks of LPS infusion were treated with neramexane for the next 2 weeksLane 7 : Rat brain whole tissue lysate - infused with LPS for 4 weeks, after 2 weeks of LPS infusion were treated with memantine for the next 2 weeks.Lysates/proteins at 40 µg per lane.SecondaryBiotinylated Goat anti-rabbit IgGdeveloped using the ECL techniquePerformed under reducing conditions.

All lanes : Anti-HMGB1 antibody - ChIP Grade (ab18256) at 1 µg/mlLane 1 : NIH 3T3 (Mouse embryonic fibroblast cell line) Whole Cell Lysate (ab7179)Lane 2 : MEF1 (Mouse embryonic fibroblast cell line) Whole Cell LysateLane 3 : PC12 (Rat adrenal pheochromocytoma cell line) Whole Cell Lysate Lysates/proteins at 10 µg per lane.SecondaryIRDye 680 Conjugated Goat Anti-Rabbit IgG (H+L) at 1/10000 dilutionPerformed under reducing conditions.

All lanes : Anti-HMGB1 antibody - ChIP Grade (ab18256) at 1 µg/mlLane 1 : HeLa (Human epithelial carcinoma cell line) Whole Cell Lysate Lane 2 : Jurkat (Human T cell lymphoblast-like cell line) Whole Cell Lysate (ab7899)Lane 3 : A431 (Human epithelial carcinoma cell line) Whole Cell Lysate (ab7909)Lane 4 : HEK293 (Human embryonic kidney cell line) Whole Cell Lysate (ab7902)Lysates/proteins at 10 µg per lane.SecondaryIRDye 680 Conjugated Goat Anti-Rabbit IgG (H+L) at 1/10000 dilutionPerformed under reducing conditions.

ICC/IF image of ab18256 stained HeLa cells. The cells were 4% PFA fixed (10 min) and then incubated in 1%BSA / 10% normal goat serum / 0.3M glycine in 0.1% PBS-Tween for 1h to permeabilise the cells and block non-specific protein-protein interactions. The cells were then incubated with the antibody (ab18256, 1µg/ml) overnight at +4°C. The secondary antibody (green) was Alexa Fluor® 488 goat anti-rabbit IgG (H+L) used at a 1/1000 dilution for 1h. Alexa Fluor® 594 WGA was used to label plasma membranes (red) at a 1/200 dilution for 1h. DAPI was used to stain the cell nuclei (blue) at a concentration of 1.43µM. This antibody also gave a positive result in 4% PFA fixed (10 min) Hek293, HepG2 and MCF7 cells at 1µg/ml.

ab18256 staining HMGB1 in Human stomach tissue sections by IHC-Fr (Immunohistochemistry - Frozen sections). Tissue samples were fixed with acetone and blocked with 5% serum for 1 hour at 25°C. Samples were incubated with primary antibody 1/500 in blocking buffer for 1 hour at 25°C. An undiluted HRP-conjugated Goat polyclonal to rabbit IgG was used as secondary antibody.See Abreview

ab18256 used in Direct ELISA in NIH 3T3 murine fibroblasts. Primary antibody used at a 1/1000 dilution for 16 hours at 4°C. The secondary antibody is an AP-conjugated Goat anti-rabbit used at a 1/1000 dilution. A blocking step was performed using 5% BSA for 1 hour.See Abreview

ICC/IF image of ab18256 stained Hela cells. The cells were 4% PFA fixed (10 min) and then incubated in 1%BSA / 10% normal goat serum / 0.3M glycine in 0.1% PBS-Tween for 1h to permeabilise the cells and block non-specific protein-protein interactions. The cells were then incubated with the antibody (ab18256, 5µg/ml) overnight at +4°C. The secondary antibody (green) was DyLight® 488 goat anti-rabbit IgG - HandL, pre-adsorbed (ab96899) used at a 1/250 dilution for 1h. Alexa Fluor® 594 WGA was used to label plasma membranes (red) at a 1/200 dilution for 1h. DAPI was used to stain the cell nuclei (blue) at a concentration of 1.43µM.

ab18256 staining HMGB1 in murine kidney tissue by Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections).Tissue was fixed in formaldehyde and a heat mediated antigen retrieval step was performed using citrate EDTA buffer pH 6.2. Samples were then blocked, then incubated with ab18256 at a 1/1000 dilution for 1 hour. The secondary used was a goat anti-rabbit IgG conjugated to HRP at a 1/1000 dilution.See Abreview

Anti-HMGB1 antibody - ChIP Grade (ab18256) at 1 µg/ml + Human HMGB1 full length protein (ab56525) at 0.01 µgSecondaryGoat Anti-Rabbit IgG H&L (HRP) preadsorbed (ab97080) at 1/5000 dilutiondeveloped using the ECL techniquePerformed under reducing conditions.Exposure time : 2 minutes

Product References

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HMGB1-promoted and TLR2/4-dependent NK cell maturation and activation take part - HMGB1-promoted and TLR2/4-dependent NK cell maturation and activation take part

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