COL1A1 was detected in immersion fixed paraffin-embedded sections of mouse Colon using Rat Anti-Mouse COL1A1, Monoclonal Antibody (Catalog #MAB11700) at 25ug/mL at 37° Celsius for 4 minutes. Before incubation with the primary antibody, tissue underwent an all-in-one dewaxing and antigen retrieval preprocessing using PreTreatment Module (PT Module) and Dewax and HIER Buffer H (pH 9; Epredia Catalog # TA-999-DHBH). Tissue was stained using the Alexa Fluor™ 647 Goat anti-Rat IgG Secondary Antibody at 1:200 at 37 ° Celsius for 2 minutes. (Yellow; Lunaphore Catalog # DR647RT) and counterstained with DAPI (blue; Lunaphore Catalog # DR100).. Specific staining was localized to connective tissue. Protocol available in COMET™ Panel Builder.

Detection of Collagen I alpha1 in Mouse Kidney.

Collagen I alpha1 was detected in perfusion fixed paraffin-embedded sections of mouse kidney using Rat Anti-Mouse Collagen I alpha1 Monoclonal Antibody (Catalog # MAB11700) at 5 µg/ml overnight at 4 °C. Before incubation with the primary antibody, tissue was subjected to heat-induced epitope retrieval using VisUCyte Antigen Retrieval Reagent-Basic (Catalog # VCTS021). Tissue was stained using the HRP-conjugated Anti-Rat IgG Secondary Antibody (Catalog # HAF005) and counterstained with hematoxylin (blue). Specific staining was localized to the cytoplasm. View our protocol for Chromogenic IHC Staining of Paraffin-embedded Tissue Sections.

Detection of Collagen I alpha1 in Mouse Pancreas.

Collagen I alpha1 was detected in perfusion fixed paraffin-embedded sections of mouse pancreas using Rat Anti-Mouse Collagen I alpha1 Monoclonal Antibody (Catalog # MAB11700) at 5 µg/ml overnight at 4 °C. Before incubation with the primary antibody, tissue was subjected to heat-induced epitope retrieval using VisUCyte Antigen Retrieval Reagent-Basic (Catalog # VCTS021). Tissue was stained using the HRP-conjugated Anti-Rat IgG Secondary Antibody (Catalog # HAF005) and counterstained with hematoxylin (blue). Specific staining was localized to the cytoplasm. View our protocol for Chromogenic IHC Staining of Paraffin-embedded Tissue Sections.

Detection of Collagen I alpha1 in 3T3-L1 cells.

Collagen I alpha1 was detected in immersion fixed 3T3-L1 mouse embryonic fibroblast adipose-like cell line (Positive) and absent in RAW 264.7 mouse monocyte/macrophage cell line (Negative) using Rat Anti-Mouse Collagen I alpha1 Monoclonal Antibody (Catalog # MAB11700) at 8 µg/ml for 3 hours at room temperature. Cells were stained using the NorthernLights™ 557-conjugated Anti-Rat IgG Secondary Antibody (red; Catalog # NL013) and counterstained with DAPI (blue). Specific staining was localized to the cytoplasm. View our protocol for Fluorescent ICC Staining of Cells on Coverslips.

Detection of Mouse Collagen I alpha1 by Western Blot.

Western Blot shows lysates of MEF mouse embryonic feeder cells. PVDF membrane was probed with 1 µg/ml of Rat Anti-Mouse Collagen I alpha1 Monoclonal Antibody (Catalog # MAB11700) followed by HRP-conjugated Anti-Rat IgG Secondary Antibody (Catalog # HAF005). A specific band was detected for Collagen I alpha1 at approximately 220 kDa (as indicated). This experiment was conducted under reducing conditions and using Western Blot Buffer Group 1.

Detection of Mouse Collagen I alpha1 by Simple Western^TM.

Simple Western lane view shows lysates of MEF mouse embryonic feeder cells, loaded at 0.1 mg/ml. A specific band was detected for Collagen I alpha1 at approximately 226 kDa (as indicated) using 10 µg/ml of Rat Anti-Mouse Collagen I alpha1 Monoclonal Antibody (Catalog # MAB11700) followed by 1:50 dilution of HRP-conjugated Anti-Rat IgG Secondary Antibody (Catalog # HAF005) in Milk-free Antibody Diluent (Catalog # 043-524). This experiment was conducted under reducing conditions and using the 66-440 kDa separation system.

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Applications for Mouse Collagen I alpha1 Antibody

Application

Recommended Usage

Immunocytochemistry

3-25 µg/mL
Sample: Immersion fixed 3T3-L1 mouse embryonic fibroblast adipose-like cell line

Immunohistochemistry

3-25 µg/mL
Sample: Perfusion fixed paraffin-embedded sections of mouse kidney and pancreas

Multiplex Immunofluorescence

25 µg/mL
Sample: Immersion fixed paraffin-embedded sections of mouse colon

Simple Western

10 µg/mL
Sample: MEF mouse embryonic feeder cells

Western Blot

1 µg/mL
Sample: MEF mouse embryonic feeder cells

Formulation, Preparation, and Storage

Purification

Protein A or G purified from hybridoma culture supernatant

Reconstitution

Reconstitute lyophilized material at 0.2 mg/ml in sterile PBS. For liquid material, refer to CoA for concentration.

Formulation

Lyophilized from a 0.2 μm filtered solution in PBS with Trehalose.

Shipping

Lyophilized product is shipped at ambient temperature. Liquid small pack size (-SP) is shipped with polar packs. Upon receipt, store immediately at the temperature recommended below.

Stability & Storage

Use a manual defrost freezer and avoid repeated freeze-thaw cycles.

12 months from date of receipt, -20 to -70 °C as supplied.
1 month, 2 to 8 °C under sterile conditions after reconstitution.
6 months, -20 to -70 °C under sterile conditions after reconstitution.

Background: Collagen I alpha 1

Type I collagen is the most abundant structural protein of connective tissues such as skin, bone and tendon. It is synthesized as a procollagen molecule which is characterized by a 300 nm triple helical domain flanked by globular N- and C-terminal propeptides (1). The triple helical domain contains Gly-Xaa-Yaa triplets where Xaa and Yaa are frequently proline and hydroxyproline, respectively. The non-helical propeptides are removed by procollagen N- and C-proteinase activities so that the mature triple helices can self-assemble into collagen fibrils that provide tensile strength to tissues (1). Type I collagen is a heterotrimer that consists of two alpha1(I) chains and one alpha2(I) chain, although homotrimers consisting of three identical alpha1(I) chains have also been described (2). This recombinant mini pro-alpha 1(I) collagen consists of a shortened alpha1(I) chain with following domain structure from N- to C-terminus: N-propeptide, N‑telopeptide, the 33 most N-terminal Gly-Xaa-Yaa repeats, the 33 most C-terminal Gly-Xaa-Yaa repeats, C-telopeptide and C-propeptide. The preparation contains a mixture of the full-length molecule, pN collagen I( alpha1) and the C-terminal propeptide. This truncated pro-alpha 1(I) collagen is a substrate for procollagen N-proteinase and procollagen C-proteinase.

References

Canty, E.G. et al. (2005) J. Cell Sci. 118:1341.
Han, S. et al. (2008) J. Mol. Biol. 383:122.

Alternate Names

COL1A1, OI4

Entrez Gene IDs

1277 (Human); 12842 (Mouse); 29393 (Rat)

Gene Symbol

COL1A1

UniProt

P11087 (Mouse)

Additional Collagen I alpha 1 Products

Product Documents for Mouse Collagen I alpha1 Antibody

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COA

SDS

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Product Specific Notices for Mouse Collagen I alpha1 Antibody

For research use only

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Mouse Collagen I alpha1 Antibody

R&D Systems, part of Bio-Techne | Catalog # MAB11700

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Product Specifications

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Scientific Data Images for Mouse Collagen I alpha1 Antibody

COL1A1 in Mouse Colon via seqIF™ staining on COMET™