Laminin alpha 3/Laminin-5 Antibody (546215) [Unconjugated]

Western blot shows lysates of human lung tissue. PVDF Membrane was probed with 2 µg/mL of Mouse Anti-Human Laminin a3/ Laminin-5 Monoclonal Antibody (Catalog # MAB21441) followed by HRP-conjugated Anti-Mouse IgG Secondary Antibody (Catalog # HAF007). A specific band was detected for Laminin a3/Laminin-5 at approximately 190 kDa (as indicated). This experiment was conducted under non-reducing conditions and using Immunoblot Buffer Group 1.

Laminin a3/Laminin-5 was detected in immersion fixed NHEK human normal epidermal keratinocytes treated with 10ng/mL Recombinant Human TGF-beta 1 (Catalog # 240-B) for 24 hours using Mouse Anti-Human Laminin a3/Laminin-5 Monoclonal Antibody (Catalog # MAB21441) at 10 µg/mL for 3 hours at room temperature. Cells were stained using the NorthernLights™ 557-conjugated Anti-Mouse IgG Secondary Antibody (red; Catalog # NL007) and counterstained with DAPI (blue). Specific staining was localized to the perinuclear space. View our protocol for Fluorescent ICC Staining of Cells on Coverslips.

Laminin alpha 3/Laminin‑5 was detected in immersion fixed paraffin-embedded sections of human stomach using Mouse Anti-Human Laminin alpha 3/Laminin‑5 Monoclonal Antibody (Catalog # MAB21441) at 5 µg/mL for 1 hour at room temperature followed by incubation with the Anti-Mouse IgG VisUCyte™ HRP Polymer Antibody (Catalog # VC001). Before incubation with the primary antibody, tissue was subjected to heat-induced epitope retrieval using Antigen Retrieval Reagent-Basic (Catalog # CTS013). Tissue was stained using DAB (brown) and counterstained with hematoxylin (blue). Specific staining was localized to cell surface of epithelial cells in gastric glands. View our protocol for IHC Staining with VisUCyte HRP Polymer Detection Reagents.

U2OS human osteosarcoma cell line was stained with Mouse Anti-Human Laminin a3/Laminin-5 Monoclonal Antibody (Catalog # MAB21441, filled histogram) or isotype control antibody (Catalog # MAB002, open histogram), followed by Allophycocyanin-conjugated Anti-Mouse IgG Secondary Antibody (Catalog # F0101B). To facilitate intracellular staining, cells were fixed with Flow Cytometry Fixation Buffer (Catalog # FC004) and permeabilized with Flow Cytometry Permeabilization/Wash Buffer I (Catalog # FC005).View our protocol for Staining Intracellular Molecules.

• Reactivity: Hu
• Applications: WB, IHC, IP, Flow, ICC/IF
• Clone: 546215
• Clonality: Monoclonal
• Host: Mouse
• Conjugate: Unconjugated
• Concentration: LYOPH

Laminin alpha 3/Laminin-5 Antibody (546215) [Unconjugated] Summary

• Immunogen: Chinese hamster ovary cell line CHO-derived recombinant human Laminin alpha 3/Laminin‑5
  aa 21-1713
  Accession # NP_000218
• Specificity: Detects human Laminin alpha 3/Laminin‑5 in direct ELISAs and Western blots. In direct ELISAs, no cross reactivity with recombinant human (rh) Laminin alpha 1, beta 1, gamma 2, or recombinant mouse Laminin alpha 4 is observed.
• Source: N/A
• Isotype: IgG1
• Clonality: Monoclonal
• Host: Mouse
• Gene: LAMA3
• Purity Statement: Protein A or G purified from hybridoma culture supernatant

Applications/Dilutions

• Dilutions:
  • Immunocytochemistry 8-25 ug/mL
  • Immunohistochemistry 5 ug/mL
  • Immunoprecipitation 25 ug/mL
  • Intracellular Staining by Flow Cytometry 0.25 ug/10^6 cells
  • Western Blot 2 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. *Small pack size (SP) is supplied either lyophilized or as a 0.2 µm filtered solution in PBS.
• Preservative: No Preservative
• Concentration: LYOPH
• Reconstitution Instructions: Reconstitute at 0.5 mg/mL in sterile PBS.

Notes

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

Alternate Names for Laminin alpha 3/Laminin-5 Antibody (546215) [Unconjugated]

• BM600 150kD subunit
• BM600
• E170
• Epiligrin 170 kDa subunit
• epiligrin alpha 3 subunit
• Epiligrin
• JEB2A
• JEB2B
• kalinin 165kD subunit
• Kalinin subunit alpha
• LAM3, alpha-3 subunit
• LAMA3
• laminin A3
• Laminin alpha 3
• laminin subunit alpha-3
• laminin-5 alpha 3 chain
• Laminin-5 subunit alpha
• Laminin5
• Laminin-5
• Laminin-6 subunit alpha
• Laminin-7 subunit alpha
• LAMNA
• LOCS
• nicein 150kD subunit
• Nicein subunit alpha

Background

Laminins are heterotrimeric, noncollagenous glycoproteins composed of alpha , beta , and gamma  chains. Through interactions with integrins, dystroglycan and other receptors, laminins contribute to cell differentiation, cell shape and migration, and maintenance of tissue phenotypes and survival. Laminin alpha 3/Laminin-5, also known as epiligrin, includes alpha 3, beta 3, and gamma 2 subunits. It is abundant in transitional epithelium, stratified squamous epithelia, lung mucosa and other epithelial glands and contributes to initiation and maintenance of epithelial cell anchorage to the underlying connective tissue. Within aa 21‑1713 of the alpha 3 subunit, human and mouse share 77% amino acid sequence identity.

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.

Publications for Laminin alpha 3/Laminin-5 Antibody (MAB21441)(5)

Publications using MAB21441

• Johannes Bischof, Oliver Patrick March, Bernadette Liemberger, Simone Alexandra Haas, Stefan Hainzl, Igor Petković et al. Paired nicking-mediated COL17A1 reframing for junctional epidermolysis bullosa Molecular Therapy 8/1/2022 [PMID: 35490295]
• Johannes Bischof, Oliver Patrick March, Bernadette Liemberger, Simone Alexandra Haas, Stefan Hainzl, Igor Petković et al. Paired nicking-mediated COL17A1 reframing for junctional epidermolysis bullosa Molecular Therapy 2022-08-01 [PMID: 35490295] (Immunocytochemistry, Human)
• E Mayr, M Ablinger, T Lettner, EM Murauer, C Guttmann-G, J Piñón Hofb, S Hainzl, M Kaiser, A Klausegger, JW Bauer, U Koller, V Wally 5'RNA Trans-Splicing Repair of COL7A1 Mutant Transcripts in Epidermolysis Bullosa International Journal of Molecular Sciences, 2022-02-02;23(3):. 2022-02-02 [PMID: 35163654] (Immunohistochemistry, Human)
• JD Howden, M Michael, W Hight-Warb, M Parsons &alpha2&beta1 integrins spatially restrict Cdc42 activity to stabilise adherens junctions Bmc Biology, 2021-06-23;19(1):130. 6/23/2021 [PMID: 34158053]
• S Furuta, G Ren, JH Mao, MJ Bissell Laminin signals initiate the reciprocal loop that informs breast-specific gene expression and homeostasis by activating NO, p53 and microRNAs Elife, 2018-03-21;7(0):. 2018-03-21 [PMID: 29560860] (IHC-P, Human)

Bioinformatics

• Gene Symbol: LAMA3
• Uniprot:
  • NP_000218()