Collagen 3 Polyclonal Antibody

$Paraformaldehyde-fixed, paraffin embedded human colon cancer; Antigen retrieval by boiling in sodium citrate buffer (pH6) for 15min; Block endogenous peroxidase by 3% hydrogen peroxide for 30 minutes; Blocking buffer (normal goat serum) at 37\u00b0C for 20min; Antibody incubation with Collagen III Polyclonal Antibody, Unconjugated (bs-0948R) at 1:200 overnight at 4\u00b0C, followed by a conjugated secondary and DAB staining.$

Paraformaldehyde-fixed, paraffin embedded human colon cancer; Antigen retrieval by boiling in sodium citrate buffer (pH6) for 15min; Block endogenous peroxidase by 3% hydrogen peroxide for 30 minutes; Blocking buffer (normal goat serum) at 37\u00b0C for 20min; Antibody incubation with Collagen III Polyclonal Antibody, Unconjugated (bs-0948R) at 1:200 overnight at 4\u00b0C, followed by a conjugated secondary and DAB staining.

Hela lysates probed with Collagen III Polyclonal Antibody, Unconjugated (bs-0948R) at 1:300 dilution and 4˚C overnight incubation. Followed by conjugated secondary antibody incubation at 1:10000 for 60 min at 37˚C.

$25 ug total protein per lane of various lysates (see on figure) probed with Collagen III polyclonal antibody, unconjugated (bs-0948R) at 1:1000 dilution and 4\u00b0C overnight incubation. Followed by conjugated secondary antibody incubation at r.t. for 60 min.$

25 ug total protein per lane of various lysates (see on figure) probed with Collagen III polyclonal antibody, unconjugated (bs-0948R) at 1:1000 dilution and 4\u00b0C overnight incubation. Followed by conjugated secondary antibody incubation at r.t. for 60 min.

$4% Paraformaldehyde-fixed RD (H) cell; Triton X-100 at r.t. for 20 min; Antibody incubation with (Collagen III) polyclonal Antibody, unconjugated (bs-0948R) 1:100, 90 min at 37\u00b0C; followed by conjugated Goat Anti-Rabbit IgG antibody (green, bs-60295G-BF488) at 37\u00b0C for 90 min, DAPI (blue, C02-04002) was used to stain the cell nuclei. PBS instead of the primary antibody was used as the blank control.$

4% Paraformaldehyde-fixed RD (H) cell; Triton X-100 at r.t. for 20 min; Antibody incubation with (Collagen III) polyclonal Antibody, unconjugated (bs-0948R) 1:100, 90 min at 37\u00b0C; followed by conjugated Goat Anti-Rabbit IgG antibody (green, bs-60295G-BF488) at 37\u00b0C for 90 min, DAPI (blue, C02-04002) was used to stain the cell nuclei. PBS instead of the primary antibody was used as the blank control.

$The RD (H) cells were fixed with 4% PFA (10 min at r.t.) and then permeabilized with 90% ice-cold methanol for 20 min at -20\u2103,the cells then were incubated in 5%BSA to block non-specific protein-protein interactions (30 min at r.t.).Primary Antibody (green):Rabbit Anti-Collagen III antibody (bs-0948R): 1 \u03bcg\/10^6 cells; Secondary Antibody (white blue): Goat anti-Rabbit IgG-BF488(bs-60295G-BF488): 1 \u03bcg\/test. Isotype Control (orange): Rabbit IgG (bs-0295P). Blank control (black): PBS. Acquisition of 20,000 events was performed.$

The RD (H) cells were fixed with 4% PFA (10 min at r.t.) and then permeabilized with 90% ice-cold methanol for 20 min at -20\u2103,the cells then were incubated in 5%BSA to block non-specific protein-protein interactions (30 min at r.t.).Primary Antibody (green):Rabbit Anti-Collagen III antibody (bs-0948R): 1 \u03bcg\/10^6 cells; Secondary Antibody (white blue): Goat anti-Rabbit IgG-BF488(bs-60295G-BF488): 1 \u03bcg\/test. Isotype Control (orange): Rabbit IgG (bs-0295P). Blank control (black): PBS. Acquisition of 20,000 events was performed.

Applications

WB
Flow-Cyt
ICC/IF

Reactivity

Human

Predicted Reactivity

Dog
Cow
Chicken

Overview
Catalog #	bs-0948r
Product Name	Collagen 3 Polyclonal Antibody
Applications	WB FLOW-CYT ICC/IF
Reactivity	Human
Predicted Reactivity	Dog, Cow, Chicken
Specifications
Conjugation	Unconjugated
Host	Rabbit
Source	KLH conjugated synthetic peptide derived from human Collagen alpha 1(III) chain
Immunogen Range	801-900/1466
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 4°C. Store at -20°C for one year. Avoid repeated freeze/thaw cycles.
Target
Gene ID	1281
Swiss Prot	P02461
Subcellular location	Secreted, Extracellular matrix
Synonyms	EDS4A; Collagen alpha-1(III) chain; COL3A1; Collagen III
Background	Collagen type III occurs in most soft connective tissues along with type I collagen. Involved in regulation of cortical development. Is the major ligand of GPR56 in the developing brain and binding to GPR56 inhibits neuronal migration and activates the RhoA pathway by coupling GPR56 to GNA13 and possibly GNA12.
Application Dilution
WB	1:500-2000
Flow-Cyt	1:100
ICC/IF	1:50-200