COL5A1 Recombinant Rabbit Monoclonal Antibody [PSH10-08]
cat.: HA723177
| Product Type: | Recombinant Rabbit monoclonal IgG, primary antibodies |
|---|---|
| Species reactivity: | Human |
| Applications: | WB, IF-Cell, FC, IP |
| Clonality: | Monoclonal |
| Clone number: | PSH10-08 |
| Form: | Liquid |
| Storage condition: | Shipped at 4℃. Store at +4℃ short term (1-2 weeks). It is recommended to aliquot into single-use upon delivery. Store at -20℃ long term. |
| Storage buffer: | PBS (pH7.4), 0.1% BSA, 40% Glycerol. Preservative: 0.05% Sodium Azide. |
| Concentration: | 1ug/ul |
| Purification: | Protein A affinity purified. |
| Molecular weight: | Predicted band size: 184 kDa |
| Isotype: | IgG |
| Immunogen: | Recombinant protein within human COL5A1 aa 251-450. |
| Positive control: | Saos-2 cell lysates, Saos-2. |
| Subcellular location: | Secreted, extracellular space, extracellular matrix. |
| Recommended Dilutions:
WB IF-Cell FC IP |
1:2,000 1:100 1:1,000 1-2μg/sample |
| Uniprot #: | SwissProt: P20908 Human |
| Alternative names: | Collagen alpha-1(V) chain COL5A1 |
Images
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Fig1:
Western blot analysis of COL5A1 on Saos-2 cell lysates with Rabbit anti-COL5A1 antibody (HA723177) at 1/2,000 dilution. Lysates/proteins at 20 µg/Lane. Predicted band size: 184 kDa Observed band size: 250 kDa Exposure time: 20 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 (HA723177) at 1/2,000 dilution was used in 5% NFDM/TBST at 4℃ overnight. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1/50,000 dilution was used for 1 hour at room temperature. |
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Fig2:
Immunocytochemistry analysis of Saos-2 cells labeling COL5A1 with Rabbit anti-COL5A1 antibody (HA723177) 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 (HA723177) at 1/100 dilution in 1% BSA in PBST overnight at 4 ℃. Goat Anti-Rabbit IgG H&L (iFluor™ 488, HA1121) 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 (HA601187, red) was stained at 1/100 dilution overnight at +4℃. Goat Anti-Mouse IgG H&L (iFluor™ 594, HA1126) was used as the secondary antibody at 1/1,000 dilution. |
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Fig3:
Flow cytometric analysis of Saos-2 cells labeling COL5A1. Cells were fixed and permeabilized. Then stained with the primary antibody (HA723177, 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 (HA1121) at 1/1,000 dilution for 30 minutes at +4℃. Unlabelled sample was used as a control (cells without incubation with primary antibody; black). |
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Fig4:
COL5A1 was immunoprecipitated from 0.2 mg Saos-2 cell lysate with HA723177 at 2 µg/10 µl beads. Western blot was performed from the immunoprecipitate using HA723177 at 1/1,000 dilution. Mouse anti Rabbit IgG heavy chain (Fc) secondary antibody (M1003-7) at 1/10,000 dilution was used for 1 hour at room temperature. Lane 1: Saos-2 cell lysate (input) Lane 2: HA723177 IP in Saos-2 cell lysate Lane 3: Rabbit IgG instead of HA723177 in Saos-2 cell lysate Blocking/Dilution buffer: 5% NFDM/TBST Exposure time: 20 seconds; ECL: K1801 |
Note: All products are “FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE”.