WB
Western blot analysis of COL1A1 on different lysates with Rabbit anti-COL1A1 antibody at 1/1,000 dilution. Lane 1: HFF-1 cell lysate (20 µg/Lane) Lane 2: NIH/3T3 cell lysate (20 µg/Lane) Lane 3: Human lung tissue lysate (40 µg/Lane) Lane 4: Mouse skin tissue lysate (40 µg/Lane) Lane 5: Rat skin tissue lysate (40 µg/Lane) Predicted band size: 139 kDa Observed band size: 200/139 kDa Exposure time: Lane 1-4: 1 minute; Lane 5: 3 seconds; ECL: K1801; 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 1/1,000 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.IHC
Immunohistochemical analysis of paraffin-embedded rat kidney tissue with Rabbit anti-COL1A1 antibody at 1/1,000 dilution. The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody at 1/1,000 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.ICC/IF
Immunocytochemistry analysis of HFF-1 cells labeling COL1A1 with Rabbit anti-COL1A1 antibody at 1/500 dilution. Cells were fixed in 4% paraformaldehyde for 20 minutes at room temperature, permeabilized with 0.1% Triton X-100 in PBS for 5 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-COL1A1 antibody at 1/500 dilution in 1% BSA in PBST overnight at 4 ℃. Goat Anti-Rabbit IgG H&L (iFluor™ 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 (iFluor™ 594) was used as the secondary antibody at 1/1,000 dilution.FC
Flow cytometric analysis of HFF-1 cells labeling COL1A1. Cells were washed twice with cold PBS and resuspend. Then stained with the primary antibody ( 1μg/mL) (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
COL1A1 was immunoprecipitated from 0.2 mg HFF-1 cell lysate with COL1A1 antibody at 2 µg/25 µl agarose. Western blot was performed from the immunoprecipitate using COL1A1 antibody at 1/1,000 dilution. Anti-Rabbit IgG for IP Nano-secondary antibody at 1/5,000 dilution was used for 1 hour at room temperature. Lane 1: HFF-1 cell lysate (input) Lane 2: COL1A1 antibody IP in HFF-1 cell lysate Lane 3: Rabbit IgG instead of COL1A1 antibody in HFF-1 cell lysate Blocking/Dilution buffer: 5% NFDM/TBST Exposure time: 8 seconds; ECL: K1802| Product Name | COL1A1 Recombinant Rabbit Monoclonal Antibody |
|---|---|
| Antibody Type | Primary Antibodies |
| Immunogen | Synthetic peptide within Human COL1A1 aa 1197-1208. |
| Clonality | Monoclonal |
|---|---|
| Isotype | IgG |
| Host Species | Rabbit |
| Tested Applications | FCICC/IFIHCIPWB |
| WB:1:1000 IHC:1:1000 ICC:1:500 FC:1:1000 IP:1:1-2μg/sample |
|
| Species Reactivity | HumanMouseRat |
| Concentration | 1mg/ml |
| Purification | Protein A |
| Gene Symbol | COL1A1 |
|---|---|
| Gene Synonyms | OI1 OI2 OI3 OI4 EDSC CAFYD EDSARTH1 |
| Gene Full Name | collagen type I alpha 1 chain |
| Gene Summary | This gene encodes the pro-alpha1 chains of type I collagen whose triple helix comprises two alpha1 chains and one alpha2 chain. Type I is a fibril-forming collagen found in most connective tissues and is abundant in bone, cornea, dermis and tendon. Mutations in this gene are associated with osteogenesis imperfecta types I-IV, Ehlers-Danlos syndrome type VIIA, Ehlers-Danlos syndrome Classical type, Caffey Disease and idiopathic osteoporosis. Reciprocal translocations between chromosomes 17 and 22, where this gene and the gene for platelet-derived growth factor beta are located, are associated with a particular type of skin tumor called dermatofibrosarcoma protuberans, resulting from unregulated expression of the growth factor. Two transcripts, resulting from the use of alternate polyadenylation signals, have been identified for this gene. [provided by R. Dalgleish, Feb 2008] |
| Molecular Weight(MW) | 139kDa |
| Cellular Localization | Secreted. |
WB
Western blot analysis of COL1A1 on different lysates with Rabbit anti-COL1A1 antibody at 1/1,000 dilution. Lane 1: HFF-1 cell lysate (20 µg/Lane) Lane 2: NIH/3T3 cell lysate (20 µg/Lane) Lane 3: Human lung tissue lysate (40 µg/Lane) Lane 4: Mouse skin tissue lysate (40 µg/Lane) Lane 5: Rat skin tissue lysate (40 µg/Lane) Predicted band size: 139 kDa Observed band size: 200/139 kDa Exposure time: Lane 1-4: 1 minute; Lane 5: 3 seconds; ECL: K1801; 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 1/1,000 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.
IHC
Immunohistochemical analysis of paraffin-embedded rat kidney tissue with Rabbit anti-COL1A1 antibody at 1/1,000 dilution. The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody at 1/1,000 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.
ICC/IF
Immunocytochemistry analysis of HFF-1 cells labeling COL1A1 with Rabbit anti-COL1A1 antibody at 1/500 dilution. Cells were fixed in 4% paraformaldehyde for 20 minutes at room temperature, permeabilized with 0.1% Triton X-100 in PBS for 5 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-COL1A1 antibody at 1/500 dilution in 1% BSA in PBST overnight at 4 ℃. Goat Anti-Rabbit IgG H&L (iFluor™ 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 (iFluor™ 594) was used as the secondary antibody at 1/1,000 dilution.
FC
Flow cytometric analysis of HFF-1 cells labeling COL1A1. Cells were washed twice with cold PBS and resuspend. Then stained with the primary antibody ( 1μg/mL) (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
COL1A1 was immunoprecipitated from 0.2 mg HFF-1 cell lysate with COL1A1 antibody at 2 µg/25 µl agarose. Western blot was performed from the immunoprecipitate using COL1A1 antibody at 1/1,000 dilution. Anti-Rabbit IgG for IP Nano-secondary antibody at 1/5,000 dilution was used for 1 hour at room temperature. Lane 1: HFF-1 cell lysate (input) Lane 2: COL1A1 antibody IP in HFF-1 cell lysate Lane 3: Rabbit IgG instead of COL1A1 antibody in HFF-1 cell lysate Blocking/Dilution buffer: 5% NFDM/TBST Exposure time: 8 seconds; ECL: K1802| Application Notes | WB:1:1000 IHC:1:1000 ICC:1:500 FC:1:1000 IP:1:1-2μg/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. |
Data sheet for OM643046