HA751058

COL1A1 Recombinant Rabbit Monoclonal Antibody [PSH06-20]

cat.: HA751058

Product Type:	Recombinant Rabbit monoclonal IgG, primary antibodies
Species reactivity:	Human, Mouse, Rat
Applications:	WB, IHC-P, IF-Tissue, IF-Cell, FC, IP, IHC-Fr
Clonality:	Monoclonal
Clone number:	PSH06-20

Form:	Liquid
Storage condition:	Store at +4℃ after thawing. Aliquot store at -20℃. Avoid repeated freeze / thaw cycles.
Storage buffer:	1*PBS (pH7.4).
Concentration:	1ug/ul
Purification:	Protein A affinity purified.
Molecular weight:	Predicted band size: 139 kDa
Isotype:	IgG
Immunogen:	Synthetic peptide within Human COL1A1 aa 1197-1208.
Positive control:	HFF-1 cell lysate, NIH/3T3 cell lysate, Human lung tissue lysate, Mouse skin tissue lysate, Rat skin tissue lysate, HFF-1, human colon cancer tissue, human kidney tissue, human lung tissue, mouse kidney tissue, mouse lung tissue, rat kidney tissue, rat lung tissue.
Subcellular location:	Secreted.
Recommended Dilutions: WB IHC-P IF-Tissue IF-Cell FC IP IHC-Fr	1:1,000-1:2,000 1:1,000 1:200 1:500 1:1,000 1-2μg/sample 1:1,000
Uniprot #:	SwissProt: P02452 Human \| P11087 Mouse \| P02454 Rat
Alternative names:	Alpha 1 type I collagen alpha1(I) procollagen CO1A1_HUMAN COL1A1 collagen alpha 1 chain type I Collagen alpha-1(I) chain collagen alpha-1(I) chain preproprotein Collagen I alpha 1 polypeptide collagen of skin, tendon and bone, alpha-1 chain Collagen type I alpha 1 EDSC OI1 OI2 OI3 OI4 pro-alpha-1 collagen type 1 type I proalpha 1 type I procollagen alpha 1 chain Type I procollagen

Images

Fig1: Western blot analysis of COL1A1 on different lysates with Rabbit anti-COL1A1 antibody (HA751058) 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 (HA751058) at 1/1,000 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.

Fig2: Western blot analysis of COL1A1 on different lysates with Rabbit anti-COL1A1 antibody (HA751058) at 1/2,000 dilution.

Lane 1: HFF-1 cell lysate (RIPA buffer)
Lane 2: HFF-1 cell lysate (Hot lysis buffer)

Predicted band size: 139 kDa
Observed band size: 200 kDa

Exposure time: 6 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 (HA751058) at 1/2,000 dilution was used in K1803 at 4℃ overnight. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1/50,000 dilution was used for 1 hour at room temperature.

	Fig3: Immunocytochemistry analysis of HFF-1 cells labeling COL1A1 with Rabbit anti-COL1A1 antibody (HA751058) 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 (HA751058) at 1/500 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 (M1305-2, 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.
	Fig4: Immunohistochemical analysis of paraffin-embedded human colon cancer tissue with Rabbit anti-COL1A1 antibody (HA751058) 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 (HA751058) 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.
	Fig5: Immunohistochemical analysis of paraffin-embedded human kidney tissue with Rabbit anti-COL1A1 antibody (HA751058) 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 (HA751058) 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.

	Fig6: Immunohistochemical analysis of paraffin-embedded human lung tissue with Rabbit anti-COL1A1 antibody (HA751058) 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 (HA751058) 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.
	Fig7: Immunohistochemical analysis of paraffin-embedded mouse kidney tissue with Rabbit anti-COL1A1 antibody (HA751058) 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 (HA751058) 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.
	Fig8: Immunohistochemical analysis of paraffin-embedded mouse lung tissue with Rabbit anti-COL1A1 antibody (HA751058) 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 (HA751058) 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.

	Fig9: Immunohistochemical analysis of paraffin-embedded rat kidney tissue with Rabbit anti-COL1A1 antibody (HA751058) 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 (HA751058) 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.
	Fig10: Immunohistochemical analysis of paraffin-embedded rat lung tissue with Rabbit anti-COL1A1 antibody (HA751058) 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 (HA751058) 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.
	Fig11: Application: IF-tissue Species: Mouse Site: Kidney Sample: Paraffin-embedded section Antibody concentration: 1/200

	Fig12: Flow cytometric analysis of HFF-1 cells labeling COL1A1. Cells were washed twice with cold PBS and resuspend. Then stained with the primary antibody (HA751058, 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 (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).
	Fig13: COL1A1 was immunoprecipitated from 0.2 mg HFF-1 cell lysate with HA751058 at 2 µg/10 µl beads. Western blot was performed from the immunoprecipitate using HA751058 at 1/1,000 dilution. Anti-Rabbit IgG for IP Nano-secondary antibody (NBI01H) at 1/5,000 dilution was used for 1 hour at room temperature. Lane 1: HFF-1 cell lysate (input) Lane 2: HA751058 IP in HFF-1 cell lysate Lane 3: Rabbit IgG instead of HA751058 in HFF-1 cell lysate Blocking/Dilution buffer: 5% NFDM/TBST Exposure time: 8 seconds; ECL: K1802
	Fig14: Application: IHC-Fr Species: Mouse Site: Lung Sample: Frozen section Antibody concentration: 1/1,000 Antigen retrieval: Not required

Note: All products are “FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE”.