WB
Western blot analysis of COL5A1 on Saos-2 cell lysates with Rabbit anti-COL5A1 antibody at 1/2,000 dilution. Lysates/proteins at 20 µg/Lane. Exposure time: 20 seconds; 4-20% SDS-PAGE gel. Proteins were transferred to a PVDF membrane and blocked with 5% NFDM/TBST for 1 hour at room temperature. The primary antibody at 1/2,000 dilution was used in 5% NFDM/TBST at 4℃ overnight. Goat Anti-Rabbit IgG - HRP Secondary Antibody at 1/50,000 dilution was used for 1 hour at room temperature.ICC/IF
Immunocytochemistry analysis of Saos-2 cells labeling COL5A1 with Rabbit anti-COL5A1 antibody at 1/100 dilution. Cells were fixed in 4% paraformaldehyde for 15 minutes at room temperature, permeabilized with 0.1% Triton X-100 in PBS for 15 minutes at room temperature, then blocked with 1% BSA in 10% negative goat serum for 1 hour at room temperature. Cells were then incubated with Rabbit anti-COL5A1 antibody at 1/100 dilution in 1% BSA in PBST overnight at 4 ℃. Goat Anti-Rabbit IgG H&L (488) was used as the secondary antibody at 1/1,000 dilution. PBS instead of the primary antibody was used as the secondary antibody only control. Nuclear DNA was labelled in blue with DAPI. Beta tubulin (red) was stained at 1/100 dilution overnight at +4℃. Goat Anti-Mouse IgG H&L (594) was used as the secondary antibody at 1/1,000 dilution.FC
Flow cytometric analysis of Saos-2 cells labeling COL5A1. Cells were fixed and permeabilized. Then stained with the primary antibody (1/1,000) (red) compared with Rabbit IgG Isotype Control (green). After incubation of the primary antibody at +4℃ for an hour, the cells were stained with a iFluor™ 488 conjugate-Goat anti-Rabbit IgG Secondary antibody at 1/1,000 dilution for 30 minutes at +4℃. Unlabelled sample was used as a control (cells without incubation with primary antibody; black).IP
COL5A1 was immunoprecipitated from 0.2 mg Saos-2 cell lysate with Rabbit anti-COL5A1 antibody at 2 µg/10 µl beads. Western blot was performed from the immunoprecipitate using Rabbit anti-COL5A1 antibody at 1/1,000 dilution. Mouse anti Rabbit IgG heavy chain (Fc) secondary antibody at 1/10,000 dilution was used for 1 hour at room temperature. Lane 1: Saos-2 cell lysate (input), Lane 2: Rabbit anti-COL5A1 antibody IP in Saos-2 cell lysate, Lane 3: Rabbit IgG instead of Rabbit anti-COL5A1 antibody in Saos-2 cell lysate. Blocking/Dilution buffer: 5% NFDM/TBST. Exposure time: 20 seconds.| Product Name | COL5A1 Recombinant Rabbit Monoclonal Antibody |
|---|---|
| Antibody Type | Primary Antibodies |
| Immunogen | Recombinant protein within human COL5A1 aa 251-450. |
| Clonality | monoclonal |
|---|---|
| Isotype | IgG |
| Host Species | Rabbit |
| Tested Applications | FCICC/IFIPWB |
| WB:1:2000 ICC/IF:1:100 FC:1:1000 IP:1-2μg/sample |
|
| Species Reactivity | Human |
| Concentration | 1mg/ml |
| Purification | Protein A |
| Gene Symbol | COL5A1 |
|---|---|
| Gene Synonyms | EDSC FMDMF EDSCL1 |
| Gene Full Name | collagen type V alpha 1 chain |
| Gene Summary | This gene encodes an alpha chain for one of the low abundance fibrillar collagens. Fibrillar collagen molecules are trimers that can be composed of one or more types of alpha chains. Type V collagen is found in tissues containing type I collagen and appears to regulate the assembly of heterotypic fibers composed of both type I and type V collagen. This gene product is closely related to type XI collagen and it is possible that the collagen chains of types V and XI constitute a single collagen type with tissue-specific chain combinations. The encoded procollagen protein occurs commonly as the heterotrimer pro-alpha1(V)-pro-alpha1(V)-pro-alpha2(V). Mutations in this gene are associated with Ehlers-Danlos syndrome, types I and II. Alternative splicing of this gene results in multiple transcript variants. [provided by RefSeq, May 2013] |
| Alternative Names | Collagen alpha-1(V) chain COL5A1 |
| Molecular Weight(MW) | 184kDa(Observed band size: 250kDa) |
| Cellular Localization | Secreted, extracellular space, extracellular matrix. |
WB
Western blot analysis of COL5A1 on Saos-2 cell lysates with Rabbit anti-COL5A1 antibody at 1/2,000 dilution. Lysates/proteins at 20 µg/Lane. Exposure time: 20 seconds; 4-20% SDS-PAGE gel. Proteins were transferred to a PVDF membrane and blocked with 5% NFDM/TBST for 1 hour at room temperature. The primary antibody at 1/2,000 dilution was used in 5% NFDM/TBST at 4℃ overnight. Goat Anti-Rabbit IgG - HRP Secondary Antibody at 1/50,000 dilution was used for 1 hour at room temperature.
ICC/IF
Immunocytochemistry analysis of Saos-2 cells labeling COL5A1 with Rabbit anti-COL5A1 antibody at 1/100 dilution. Cells were fixed in 4% paraformaldehyde for 15 minutes at room temperature, permeabilized with 0.1% Triton X-100 in PBS for 15 minutes at room temperature, then blocked with 1% BSA in 10% negative goat serum for 1 hour at room temperature. Cells were then incubated with Rabbit anti-COL5A1 antibody at 1/100 dilution in 1% BSA in PBST overnight at 4 ℃. Goat Anti-Rabbit IgG H&L (488) was used as the secondary antibody at 1/1,000 dilution. PBS instead of the primary antibody was used as the secondary antibody only control. Nuclear DNA was labelled in blue with DAPI. Beta tubulin (red) was stained at 1/100 dilution overnight at +4℃. Goat Anti-Mouse IgG H&L (594) was used as the secondary antibody at 1/1,000 dilution.
FC
Flow cytometric analysis of Saos-2 cells labeling COL5A1. Cells were fixed and permeabilized. Then stained with the primary antibody (1/1,000) (red) compared with Rabbit IgG Isotype Control (green). After incubation of the primary antibody at +4℃ for an hour, the cells were stained with a iFluor™ 488 conjugate-Goat anti-Rabbit IgG Secondary antibody at 1/1,000 dilution for 30 minutes at +4℃. Unlabelled sample was used as a control (cells without incubation with primary antibody; black).
IP
COL5A1 was immunoprecipitated from 0.2 mg Saos-2 cell lysate with Rabbit anti-COL5A1 antibody at 2 µg/10 µl beads. Western blot was performed from the immunoprecipitate using Rabbit anti-COL5A1 antibody at 1/1,000 dilution. Mouse anti Rabbit IgG heavy chain (Fc) secondary antibody at 1/10,000 dilution was used for 1 hour at room temperature. Lane 1: Saos-2 cell lysate (input), Lane 2: Rabbit anti-COL5A1 antibody IP in Saos-2 cell lysate, Lane 3: Rabbit IgG instead of Rabbit anti-COL5A1 antibody in Saos-2 cell lysate. Blocking/Dilution buffer: 5% NFDM/TBST. Exposure time: 20 seconds.| Application Notes | WB:1:2000 ICC/IF:1:100 FC:1:1000 IP:1-2μg/sample |
|---|
| Form | Liquid |
|---|---|
| Storage Instructions | Store at +4℃ after thawing. Aliquot store at -20℃. Avoid repeated freeze / thaw cycles. |
| Storage Buffer | PBS (pH7.4), 0.1% BSA, 40% Glycerol. Preservative: 0.05% Sodium Azide. |
