Anti-HMGB1 antibody

产品名称

Anti-HMGB1抗体
参阅全部 HMGB1 一抗

描述

兔多克隆抗体to HMGB1

宿主

Rabbit

经测试应用

适用于: ICC/IF, WBmore details

种属反应性

与反应: Mouse, Rat, Human
预测可用于: Rabbit, Cow

免疫原

Synthetic peptide. This information is proprietary to Abcam and/or its suppliers.

阳性对照

常规说明

The Life Science industry has been in the grips of a reproducibility crisis for a number of years. Abcam is leading the way in addressing this with our range of recombinant monoclonal antibodies and knockout edited cell lines for gold-standard validation. Please check that this product meets your needs before purchasing.

If you have any questions, special requirements or concerns, please send us an inquiry and/or contact our Support team ahead of purchase. Recommended alternatives for this product can be found below, along with publications, customer reviews and Q&As

形式

Liquid

存放说明

Shipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C or -80°C. Avoid freeze / thaw cycle.

存储溶液

pH: 7.40
Preservative: 0.02% Sodium azide
Constituents: 98.98% PBS, 1% BSA

Batches of this product that have a concentration < 1mg/ml may have BSA added as a stabilising agent. If you would like information about the formulation of a specific lot, please contact our scientific support team who will be happy to help.

浓度

纯度

Immunogen affinity purified

克隆

多克隆

同种型

IgG

The Abpromise guarantee

Abpromise™承诺保证使用ab18256于以下的经测试应用

“应用说明”部分 下显示的仅为推荐的起始稀释度；实际最佳的稀释度/浓度应由使用者检定。

数据库链接

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别名

Immunocytochemistry/ Immunofluorescence - Anti-HMGB1 antibody (ab18256)

ab18256 staining HMGB1 in HeLa cells. The cells were fixed with 4% paraformaldehyde (10 min), permeabilized with 0.1% PBS-Triton X-100 for 5 minutes and then blocked with 1% BSA/10% normal goat serum/0.3M glycine in 0.1%PBS-Tween for 1h. The cells were then incubated overnight at 4°C with ab18256 at 1µg/ml and ab7291, Mouse monoclonal [DM1A] to alpha Tubulin - Loading Control. Cells were then incubated with ab150081, Goat polyclonal Secondary Antibody to Rabbit IgG - H&L (Alexa Fluor® 488), pre-adsorbed at 1/1000 dilution (shown in green) and ab150120, Goat polyclonal Secondary Antibody to Mouse IgG - H&L (Alexa Fluor® 594), pre-adsorbed at 1/1000 dilution (shown in pseudocolour magenta). Nuclear DNA was labelled with DAPI (shown in blue).

Image was acquired with a high-content analyser (Operetta CLS, Perkin Elmer) and a maximum intensity projection of confocal sections is shown.

Western blot - Anti-HMGB1 antibody (ab18256)

All lanes : Anti-HMGB1 antibody (ab18256) at 1 µg/ml

Lane 1 : Wild-type HAP1 whole cell lysate
Lane 2 : HMGB1 knockout HAP1 whole cell lysate

Lysates/proteins at 20 µg per lane.

Predicted band size: 25 kDa

Lanes 1 - 4: Merged signal (red and green). Green - ab18256 observed at 29 kDa. Red - loading control, ab9484, observed at 37 kDa.

ab18256 was shown to recognize HMGB1 in wild-type HAP1 cells as signal was lost at the expected MW in HMGB1 knockout cells. Additional cross-reactive bands were observed in the wild-type and knockout cells. Wild-type and HMGB1 knockout samples were subjected to SDS-PAGE. Ab18256 and ab9484 (Mouse anti-GAPDH loading control) were incubated overnight at 4°C at 1 μg/ml and 1/20000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye® 800CW) preabsorbed ab216773 and Goat anti-Mouse IgG H&L (IRDye® 680RD) preabsorbed ab216776 secondary antibodies at 1/20000 dilution for 1 hour at room temperature before imaging.

Western blot - Anti-HMGB1 antibody (ab18256)

All lanes : Anti-HMGB1 antibody (ab18256) at 1 µg/ml

Lane 1 : NIH/3T3 whole cell lysate (ab7179)
Lane 2 : MEF1 (Mouse embryonic fibroblast cell line) Whole Cell Lysate
Lane 3 : PC-12 (Rat adrenal pheochromocytoma cell line) Whole Cell Lysate

Lysates/proteins at 10 µg per lane.

Secondary
Lanes 1 & 3 : IRDye 680 Conjugated Goat Anti-Rabbit IgG (H+L) at 1/10000 dilution
Lane 2 : IRDye 680 Conjugated Goat Anti-Rabbit IgG (H+L) at 1/10000 dilution

Performed under reducing conditions.

Predicted band size: 25 kDa
Observed band size: 29 kDawhy is the actual band size different from the predicted?
Additional bands at: 59 kDa. We are unsure as to the identity of these extra bands.

Western blot - Anti-HMGB1 antibody (ab18256)

All lanes : Anti-HMGB1 antibody (ab18256) at 1 µg/ml

Lane 1 : HeLa (Human epithelial cell line from cervix adenocarcinoma) whole cell lysate
Lane 2 : Jurkat whole cell lysate (ab7899)
Lane 3 : A-431 whole cell lysate (ab7909)
Lane 4 : HEK-293 whole cell lysate (ab7902)

Lysates/proteins at 10 µg per lane.

Secondary
All lanes : IRDye 680 Conjugated Goat Anti-Rabbit IgG (H+L) at 1/10000 dilution

Performed under reducing conditions.

Predicted band size: 25 kDa
Observed band size: 29 kDawhy is the actual band size different from the predicted?

Western blot - Anti-HMGB1 antibody (ab18256)

Anti-HMGB1 antibody (ab18256) at 1 µg/ml + Recombinant Human HMGB1 protein (ab56525) at 0.01 µg

Secondary
Goat Anti-Rabbit IgG H&L (HRP) preadsorbed (ab97080) at 1/5000 dilution

Developed using the ECL technique.

Performed under reducing conditions.

Predicted band size: 25 kDa


Exposure time: 2 minutes

Western blot - Anti-HMGB1 antibody (ab18256)This image is courtesy of an Abreview submitted by Dr Francesca Cerbai

All lanes : Anti-HMGB1 antibody (ab18256) at 1/1000 dilution

Lane 1 : Rat brain whole tissue lysate - infused with asf for 1 week
Lane 2 : Rat brain whole tissue lysate - infused with LPS for 1 week
Lane 3 : Rat brain whole tissue lysate - infused with acsf for 8 weeks
Lane 4 : Rat brain whole tissue lysate - infused with LPS for 8 weeks
Lane 5 : Rat brain whole tissue lysate - infused with LPS for 4 weeks
Lane 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 weeks
Lane 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.

Secondary
All lanes : Biotinylated Goat anti-rabbit IgG

Developed using the ECL technique.

Performed under reducing conditions.

Predicted band size: 25 kDa


Exposure time: 30 seconds

See Abreview