Collagen I alpha 1 Antibody (1102924) [Unconjugated]

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.​

Collagen I alpha 1 was detected in perfusion fixed paraffin-embedded sections of mouse kidney using Rat Anti-Mouse Collagen I alpha 1 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.

Collagen I alpha 1 was detected in perfusion fixed paraffin-embedded sections of mouse pancreas using Rat Anti-Mouse Collagen I alpha 1 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.

Collagen I alpha 1 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 alpha 1 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.

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

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 alpha 1 at approximately 226 kDa (as indicated) using 10 µg/ml of Rat Anti-Mouse Collagen I alpha 1 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.

• Reactivity: Mu
• Applications: WB, Simple Western, IHC, ICC/IF, mIF
• Clone: 1102924
• Clonality: Monoclonal
• Host: Rat
• Conjugate: Unconjugated

Collagen I alpha 1 Antibody (1102924) [Unconjugated] Summary

• Immunogen: Synthetic Peptide
  Accession # P11087
• Specificity: Detects a synthetic peptide specific for mouse COL1A1 around amino acid 215 in Direct ELISA.
• Source: N/A
• Isotype: IgG2b
• Clonality: Monoclonal
• Host: Rat
• Purity Statement: Protein A or G purified from hybridoma culture supernatant

Applications/Dilutions

• Dilutions:
  • Immunocytochemistry 3-25 ug/mL
  • Immunohistochemistry 3-25 ug/mL
  • Multiplex Immunofluorescence 25 ug/mL
  • Simple Western 10 ug/mL
  • Western Blot 1 ug/mL

Packaging, Storage & Formulations

• 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.
• Buffer: Lyophilized from a 0.2 μm filtered solution in PBS with Trehalose.
• Reconstitution Instructions: Reconstitute lyophilized material at 0.2 mg/ml in sterile PBS. For liquid material, refer to CoA for concentration.

Notes

This product is produced by and ships from R&D Systems, Inc., a Bio-Techne brand.

Alternate Names for Collagen I alpha 1 Antibody (1102924) [Unconjugated]

• Alpha-1 type I collagen
• COL1A1
• COL-IA1
• Collagen 1 alpha 1
• Collagen 1
• collagen alpha 1 chain type I
• collagen alpha-1(I) chain
• Collagen I alpha 1
• collagen of skin, tendon and bone, alpha-1 chain
• Collagen Type I Alpha 1 Chain
• collagen, type I, alpha 1
• Collagen1
• Collagen-1
• EDSARTH1
• OI4
• pro-alpha-1 collagen type 1
• Type I Procollagen Alpha 1 Chain

Background

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 alpha 1(I) chains and one alpha 2(I) chain, although homotrimers consisting of three identical alpha 1(I) chains have also been described (2). This recombinant mini pro-alpha 1(I) collagen consists of a shortened alpha 1(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( alpha 1) and the C-terminal propeptide. This truncated pro-alpha 1(I) collagen is a substrate for procollagen N-proteinase and procollagen C-proteinase.

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

Limitations

This product is for research use only and is not approved for use in humans or in clinical diagnosis. Primary Antibodies are guaranteed for 1 year from date of receipt.

FAQs for Collagen I alpha 1 Antibody (MAB11700). (Showing 1 - 4 of 4 FAQ).

1. We are looking for an anti-Collagen antibody for Flow Cytometry (FACS) analysis in Swine sample. Would you please provide some references if there is any available as well?
   • We currently have one antibody for porcine Collagen validated in flow cytometry (catalog # NBP1-94202).
2. I am looking for anti collagen 1 antibody, I see your product calls it collagen I alpha 1 is there a difference?
   • Collagen I alpha 1 is the most common protein for the collagen 1 protein. There are other subunits as well (such as collagen 1 alpha 2…etc.).
3. The predicted molecular weight says 150kDA but the band on the gel appears at 130kDA why is that?
   • Differences between the predicted and observed MW can be affected by a number of factors including PTMs, relative charges and experimental factors. These cumulative effect of these differences can be quite substantial with larger molecules such as collagen I alpha 1.
4. How long does delivery take for your collagen 1 alpha 1 antibody?
   • Our most popular collagen I alpha 1 antibody (NB600-408) is usually in stock and ships priority overnight for next business day delivery in the U.S.

Bioinformatics

• Uniprot:
  • P11087(Mouse)