Human Collagen I  alpha 1 Antibody

Recombinant Monoclonal Antibody
Catalog # Availability Size / Price Qty
MAB11757-100
MAB11757-SP

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Detection of Col1A1 in Human Colon Cancer via seqIF™ staining on COMET™​
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Human Collagen I  alpha 1 Antibody Summary

Species Reactivity
Human
Specificity
Detects recombinant human COL1 alpha 1 protein in Direct ELISA.
Source
Recombinant Monoclonal Rabbit IgG Clone # 3158C
Purification
Protein A or G purified from hybridoma culture supernatant
Immunogen
Chinese Hamster Ovary-Derived recombinant human COL-1A1
Gln23-Lys277, Gly1094-Leu1464
Accession # P02452
Formulation
Lyophilized from a 0.2 μm filtered solution in PBS with Trehalose.
Label
Unconjugated

Applications

Recommended Concentration
Sample
Western Blot
2 µg/mL
Saos-2 human osteosarcoma cell line and IMR-90 human lung fibroblast cell line
Immunohistochemistry
1-10 µg/mL
Immersion fixed paraffin-embedded sections of human breast tumor, human cervix, human colon cancer, and human colon
Multiplex Immunofluorescence
20 µg/mL
Immersion fixed paraffin-embedded sections of human Colon Cancer

Please Note: Optimal dilutions should be determined by each laboratory for each application. General Protocols are available in the Technical Information section on our website.

Scientific Data

Multiplex Immunofluorescence View Larger

Detection of Col1A1 in Human Colon Cancer via seqIF™ staining on COMET™​ Col1A1 was detected in immersion fixed paraffin-embedded sections of human Colon Cancer using Rabbit Anti-Human Col1A1, Monoclonal Antibody (Catalog # MAB) at 20ug/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™ Plus 647 Goat anti-Rabbit IgG Secondary Antibody at 1:200 at 37 ° Celsius for 2 minutes. (Yellow; Lunaphore Catalog # DR647RB) and counterstained with DAPI (blue; Lunaphore Catalog # DR100). Specific staining was localized to the cytoplasm. Protocol available in COMET™ Panel Builder.

Western Blot View Larger

Detection of Human Collagen I  alpha 1 by Western Blot. Western Blot shows lysates of Saos‑2 human osteosarcoma cell line and IMR‑90 human lung fibroblast cell line. PVDF membrane was probed with 1 µg/ml of Rabbit Anti-Human Collagen I  alpha 1 Monoclonal Antibody (Catalog # MAB11757) followed by HRP-conjugated Anti-Rabbit IgG Secondary Antibody (Catalog # HAF008). 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.

Immunohistochemistry View Larger

Detection of Collagen I  alpha 1 in Human Breast Tumor. Collagen I  alpha 1 was detected in immersion fixed paraffin-embedded sections of human breast tumor using Rabbit Anti-Human Collagen I  alpha 1 Monoclonal Antibody (Catalog # MAB11757) at 3 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-Rabbit IgG Secondary Antibody (Catalog # HAF008) and counterstained with hematoxylin (blue). Specific staining was localized to the cytoplasm. View our protocol for Chromogenic IHC Staining of Paraffin-embedded Tissue Sections.

Immunohistochemistry View Larger

Detection of Collagen I  alpha 1 in Human Cervix. Collagen I  alpha 1 was detected in immersion fixed paraffin-embedded sections of human cervix using Rabbit Anti-Human Collagen I  alpha 1 Monoclonal Antibody (Catalog # MAB11757) at 3 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-Rabbit IgG Secondary Antibody (Catalog # HAF008) and counterstained with hematoxylin (blue). Specific staining was localized to the cytoplasm. View our protocol for Chromogenic IHC Staining of Paraffin-embedded Tissue Sections.

Immunohistochemistry View Larger

Detection of Collagen I  alpha 1 in Human Colon Cancer. Collagen I  alpha 1 was detected in immersion fixed paraffin-embedded sections of human colon cancer using Rabbit Anti-Human Collagen I  alpha 1 Monoclonal Antibody (Catalog # MAB11757) at 3 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-Rabbit IgG Secondary Antibody (Catalog # HAF008) and counterstained with hematoxylin (blue). Specific staining was localized to the cytoplasm. View our protocol for Chromogenic IHC Staining of Paraffin-embedded Tissue Sections.

Immunohistochemistry View Larger

Detection of Collagen I  alpha 1 in Human Colon. Collagen I  alpha 1 was detected in immersion fixed paraffin-embedded sections of human colon using Rabbit Anti-Human Collagen I  alpha 1 Monoclonal Antibody (Catalog # MAB11757) at 3 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-Rabbit IgG Secondary Antibody (Catalog # HAF008) and counterstained with hematoxylin (blue). Specific staining was localized to the cytoplasm. View our protocol for Chromogenic IHC Staining of Paraffin-embedded Tissue Sections.

Reconstitution Calculator

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Preparation and Storage

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

References
  1. Canty, E.G. et al. (2005) J. Cell Sci. 118:1341.
  2. Han, S. et al. (2008) J. Mol. Biol. 383:122.
Entrez Gene IDs
1277 (Human); 12842 (Mouse); 29393 (Rat)
Alternate Names
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

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