HMGB1 antibody

GTX101277_41885_20160602_WB_shRNA_watermark_19011506_129.jpg

Cat No. GTX101277

Reference 14

Host	Rabbit
Clonality	Polyclonal
Isotype	IgG
Application	WB, ICC/IF, IHC-P, IHC
Reactivity	Human, Mouse, Rat, Pig

See all HMGB1 products

APPLICATION

Application Note

*Optimal dilutions/concentrations should be determined by the researcher.

Application	Dilution
WB	1:500-1:3000
ICC/IF	1:100-1:1000
IHC-P	1:100-1:1000
IHC	Assay dependent

Not tested in other applications.

Calculated MW

25 kDa. ( Note )

Positive Control

293T , A431 , HeLa , HepG2 , NIH-3T3 , JC , BCL-1 , C2C12 , Raw264.7 , PC-12 , HMGB1-transfected 293T

Predict Reactivity

Rabbit, Bovine, Dog, Chicken, Xenopus laevis, Rhesus Monkey(>80% identity)

PROPERTIES

Form

Liquid

Buffer

1XPBS, 1% BSA, 20% Glycerol (pH7). 0.025% ProClin 300 was added as a preservative.

Storage

Store as concentrated solution. Centrifuge briefly prior to opening vial. For short-term storage (1-2 weeks), store at 4ºC. For long-term storage, aliquot and store at -20ºC or below. Avoid multiple freeze-thaw cycles.

Concentration

0.43 mg/ml (Please refer to the vial label for the specific concentration.)

Antigen Species

Human

Immunogen

Recombinant protein encompassing a sequence within the center region of human HMGB1. The exact sequence is proprietary.

Purification

Purified by antigen-affinity chromatography.

Conjugation

Unconjugated

Note

For laboratory use only. Not for any clinical, therapeutic, or diagnostic use in humans or animals. Not for animal or human consumption.

TARGET

Synonyms

High Mobility Group Box 1,Hmg-1,Hmg1,Hmg3,Sbp-1,Hmgb1

Cellular Localization

Nucleus

Background

DNA binding proteins that associates with chromatin and has the ability to bend DNA. Binds preferentially single-stranded DNA. Involved in V(D)J recombination by acting as a cofactor of the RAG complex. Acts by stimulating cleavage and RAG protein binding at the 23 bp spacer of conserved recombination signal sequences (RSS). Heparin-binding protein that has a role in the extension of neurite-type cytoplasmic processes in developing cells.

Database

Gene ID: 3146
UniProt: P09429

Research Area

DATA IMAGES

GTX101277 WB Image

Non-transfected (–) and transfected (+) 293T whole cell extracts (30 µg) were separated by 12% SDS-PAGE, and the membrane was blotted with HMGB1 antibody (GTX101277) diluted at 1:5000. The HRP-conjugated anti-rabbit IgG antibody (GTX213110-01) was used to detect the primary antibody.

GTX101277 ICC/IF Image

HMGB1 antibody detects HMGB1 protein at nucleus by immunofluorescent analysis.
Sample: HeLa cells were fixed in 4% paraformaldehyde at RT for 15 min.
Green: HMGB1 protein stained by HMGB1 antibody (GTX101277) diluted at 1:1000.
Red: phalloidin, a cytoskeleton marker, stained by phalloidin (invitrogen, A12380) diluted at 1:200.
Blue: Hoechst 33342 staining.

GTX101277 WB Image

HMGB1 antibody detects HMGB1 protein by western blot analysis.
A. 30 µg NIH-3T3 whole cell lysate/extract
B. 30 µg JC whole cell lysate/extract
C. 30 µg BCL-1 whole cell lysate/extract
D. 30 µg C2C12 whole cell lysate/extract
E. 30 µg Raw264.7 whole cell lysate/extract
12% SDS-PAGE
HMGB1 antibody (GTX101277) dilution: 1:3000
The HRP-conjugated anti-rabbit IgG antibody (GTX213110-01) was used to detect the primary antibody.

GTX101277 IHC-P Image

HMGB1 antibody detects HMGB1 protein at nucleus on mouse colon by immunohistochemical analysis.
Sample: Paraffin-embedded mouse colon.
HMGB1 antibody (GTX101277) dilution: 1:1000.

Antigen Retrieval: Trilogy™ (EDTA based, pH 8.0) buffer, 15min

GTX101277 WB Image

HMGB1 antibody detects HMGB1 protein by western blot analysis.
A. 30 µg PC-12 whole cell lysate/extract
12% SDS-PAGE
HMGB1 antibody (GTX101277) dilution: 1:3000
The HRP-conjugated anti-rabbit IgG antibody (GTX213110-01) was used to detect the primary antibody.

GTX101277 ICC/IF Image

HMGB1 antibody detects HMGB1 protein at nucleus by immunofluorescent analysis.
Sample: NIH/3T3 cells were fixed in 4% paraformaldehyde at RT for 15 min.
Green: HMGB1 protein stained by HMGB1 antibody (GTX101277) diluted at 1:500.
Red: phalloidin, a cytoskeleton marker, diluted at 1:50.
Scale bar = 10 μm.

GTX101277 IHC-P Image

HMGB1 antibody detects HMGB1 protein at nucleus in mouse esophagus by immunohistochemical analysis.
Sample: Paraffin-embedded mouse esophagus.
HMGB1 antibody (GTX101277) diluted at 1:1000.

Antigen Retrieval: Citrate buffer, pH 6.0, 15 min

GTX101277 WB Image

Various whole cell extracts (30 µg) were separated by 12% SDS-PAGE, and the membrane was blotted with HMGB1 antibody (GTX101277) diluted at 1:3000. The HRP-conjugated anti-rabbit IgG antibody (GTX213110-01) was used to detect the primary antibody.

GTX101277 WB Image

GTX101277 IHC-P Image

HMGB1 antibody detects HMGB1 protein at nucleus on mouse esophagus by immunohistochemical analysis.
Sample: Paraffin-embedded mouse esophagus.
HMGB1 antibody (GTX101277) dilution: 1:1000.

Antigen Retrieval: Trilogy™ (EDTA based, pH 8.0) buffer, 15min

GTX101277 ICC/IF Image

HMGB1 antibody detects HMGB1 protein at cytoplasm and nucleus by immunofluorescent analysis.
Sample: SK-N-SH cells were fixed in 4% paraformaldehyde at RT for 15 min.
Green: HMGB1 protein stained by HMGB1 antibody (GTX101277) diluted at 1:1000.
Red: beta Tubulin 3/ Tuj1, a cytoskeleton marker, stained by beta Tubulin 3/ Tuj1 antibody [GT11710] (GTX631836) diluted at 1:500.
Scale bar = 10 μm.

GTX101277 IHC-P Image

HMGB1 antibody detects HMGB1 protein at nucleus on rat brain stem by immunohistochemical analysis.
Sample: Paraffin-embedded rat brain stem.
HMGB1 antibody (GTX101277) dilution: 1:1000.

Antigen Retrieval: Trilogy™ (EDTA based, pH 8.0) buffer, 15min

REFERENCE