HMGB1 Polyclonal Antibody

Formalin-fixed and paraffin embedded human endometrium carcinoma labeled Anti-HMGB1 Polyclonal Antibody, Unconjugated (bs-0664R) at 1:200, followed by conjugation to the secondary antibody and DAB staining

Lane 1: Mouse Macrophages probed with Rabbit Anti-HMGB1 Polyclonal Antibody, Unconjugated (bs-0664R) at 1:300 overnight at 4˚C. Followed by conjugation to secondary antibody (bs-0295G-HRP) at 1:5000 for 90 min at RT.

Human HepG2 cell lysates probed with Rabbit Anti-HMGB1 Polyclonal Antibody, PE-Cy5 conjugated (bs-0664R-PE-Cy5) (green) at 1:20 for 30 minutes followed compared to unstained cells (blue) and PE-Cy5 isotype control bs-0295P-PE-Cy5(orange).

$HL-60 cells were fixed with 4% PFA for 10min at room temperature,permeabilized with 90% ice-cold methanol for 20 min at -20\u2103, and incubated in 5% BSA blocking buffer for 30 min at room temperature. Cells were then stained with HMGB1 Polyclonal Antibody(bs-0664R)at 1:100 dilution in blocking buffer and incubated for 30 min at room temperature, washed twice with 2%BSA in PBS, followed by secondary antibody incubation for 40 min at room temperature. Acquisitions of 20,000 events were performed. Cells stained with primary antibody (green), and isotype control (orange).$

HL-60 cells were fixed with 4% PFA for 10min at room temperature,permeabilized with 90% ice-cold methanol for 20 min at -20\u2103, and incubated in 5% BSA blocking buffer for 30 min at room temperature. Cells were then stained with HMGB1 Polyclonal Antibody(bs-0664R)at 1:100 dilution in blocking buffer and incubated for 30 min at room temperature, washed twice with 2%BSA in PBS, followed by secondary antibody incubation for 40 min at room temperature. Acquisitions of 20,000 events were performed. Cells stained with primary antibody (green), and isotype control (orange).

$HepG2 cell; 4% Paraformaldehyde-fixed; Triton X-100 at room temperature for 20 min; Blocking buffer (normal goat serum, C-0005) at 37\u00b0C for 20 min; Antibody incubation with (HMGB1) polyclonal Antibody, Unconjugated (bs-0664R) 1:100, 90 minutes at 37\u00b0C; followed by a conjugated Goat Anti-Rabbit IgG antibody at 37\u00b0C for 90 minutes, DAPI (blue, C02-04002) was used to stain the cell nuclei.$

HepG2 cell; 4% Paraformaldehyde-fixed; Triton X-100 at room temperature for 20 min; Blocking buffer (normal goat serum, C-0005) at 37\u00b0C for 20 min; Antibody incubation with (HMGB1) polyclonal Antibody, Unconjugated (bs-0664R) 1:100, 90 minutes at 37\u00b0C; followed by a conjugated Goat Anti-Rabbit IgG antibody at 37\u00b0C for 90 minutes, DAPI (blue, C02-04002) was used to stain the cell nuclei.

Applications

WB
ELISA
FCM
IHC-P
IHC-F
IF(IHC-P)
IF(IHC-F)
IF(ICC)

Reactivity

Human
Mouse
Rat

Predicted Reactivity

Overview
Catalog #	bs-0664R
Product Name	HMGB1 Polyclonal Antibody
Applications	WB ELISA FCM IHC-P IHC-F IF(IHC-P) IF(IHC-F) IF(ICC)
Reactivity	Human, Mouse, Rat
Predicted Reactivity	Cow
Specifications
Conjugation	Unconjugated
Host	Rabbit
Source	KLH conjugated synthetic peptide derived from human HMGB1
Clonality	Polyclonal
Isotype	IgG
Concentration	1ug/ul
Purification	Purified by Protein A.
Storage Buffer	0.01M TBS(pH7.4) with 1% BSA, 0.02% Proclin300 and 50% Glycerol.
Storage Condition	Shipped at 4C. Store at -20C for one year. Avoid repeated freeze/thaw cycles.
Target
Gene ID	3146
Swiss Prot	P09429
Subcellular location	Nucleus
Synonyms	HMG1; HMG3; SBP-1; High mobility group protein B1; High mobility group protein 1; HMG-1; HMGB1
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 (By similarity).
Application Dilution
WB	1:300-5000
ELISA	1:500-1000
FCM	1:20-100
IHC-P	1:200-400
IHC-F	1:100-500
IF(IHC-P)	1:50-200
IF(IHC-F)	1:50-200
IF(ICC)	1:50-200