Alpha-2-macroglobulin Mouse Monoclonal Antibody [A5-A9]
cat.: M1510-22
Product Type: Mouse monoclonal IgG1, primary antibodies
Species reactivity: Human
Applications: WB, IF-Cell, IHC-P, ELISA
Clonality: Monoclonal
Clone number: A5-A9
Form: Liquid
Storage condition: Store at +4℃ after thawing. Aliquot store at -20℃ or -80℃. Avoid repeated freeze / thaw cycles.
Storage buffer: 1*PBS (pH7.4), 0.2% BSA, 40% Glycerol. Preservative: 0.05% Sodium Azide.
Concentration: 2ug/ul
Purification: Protein G affinity purified.
Molecular weight: Predicted band size: 163 kDa
Isotype: IgG1
Immunogen: Recombinant protein within human Alpha-2-macroglobulin aa 950-1200.
Positive control: LOVO cell lysates, LO2 cell lysates, Hela, HepG2, human liver tissue, human uterus tissue.
Subcellular location: Secreted.
Recommended Dilutions:
  WB
  IF-Cell
  IHC-P
  ELISA
1:500-1:1,000
1:50-1:200
1:100-1:500
1:1,000-1:10,000
Uniprot #: SwissProt: P01023 Human
Alternative names: A2m A2MG_HUMAN Alpha 2 M Alpha 2M Alpha-2-M Alpha-2-macroglobulin C3 and PZP-like alpha-2-macroglobulin domain-containing protein 5 CPAMD5 DKFZp779B086 FWP007 S863 7
Images
M1510-22_1.jpg Fig1: Western blot analysis of Alpha-2-macroglobulin on LOVO cell lysate. Proteins were transferred to a PVDF membrane and blocked with 5% BSA in PBS for 1 hour at room temperature. The primary antibody (M1510-22, 1/500) was used in 5% BSA at room temperature for 2 hours. Goat Anti-Mouse IgG - HRP Secondary Antibody (HA1006) at 1:5,000 dilution was used for 1 hour at room temperature.
M1510-22_2.jpg Fig2: Western blot analysis of Alpha-2-macroglobulin on LO2 cell lysate. Proteins were transferred to a PVDF membrane and blocked with 5% BSA in PBS for 1 hour at room temperature. The primary antibody (M1510-22, 1/500) was used in 5% BSA at room temperature for 2 hours. Goat Anti-Mouse IgG - HRP Secondary Antibody (HA1006) at 1:5,000 dilution was used for 1 hour at room temperature.
M1510-22_3.jpg Fig3: ICC staining of Alpha-2-macroglobulin in Hela cells (green). Formalin fixed cells were permeabilized with 0.1% Triton X-100 in TBS for 10 minutes at room temperature and blocked with 1% Blocker BSA for 15 minutes at room temperature. Cells were probed with the primary antibody (M1510-22, 1/50) for 1 hour at room temperature, washed with PBS. Alexa Fluor®488 Goat anti-Mouse IgG was used as the secondary antibody at 1/1,000 dilution. The nuclear counter stain is DAPI (blue).
M1510-22_4.jpg Fig4: ICC staining of Alpha-2-macroglobulin in HepG2 cells (green). Formalin fixed cells were permeabilized with 0.1% Triton X-100 in TBS for 10 minutes at room temperature and blocked with 1% Blocker BSA for 15 minutes at room temperature. Cells were probed with the primary antibody (M1510-22, 1/100) for 1 hour at room temperature, washed with PBS. Alexa Fluor®488 Goat anti-Mouse IgG was used as the secondary antibody at 1/1,000 dilution. The nuclear counter stain is DAPI (blue).
M1510-22_5.jpg Fig5: Immunohistochemical analysis of paraffin-embedded human liver tissue using anti-Alpha-2-macroglobulin antibody. The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 8.0-8.4) for 20 minutes.The tissues were blocked in 5% BSA for 30 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (M1510-22, 1/500) for 30 minutes 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.
M1510-22_6.jpg Fig6: Immunohistochemical analysis of paraffin-embedded human uterus tissue using anti-Alpha-2-macroglobulin antibody. The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 8.0-8.4) for 20 minutes.The tissues were blocked in 5% BSA for 30 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (M1510-22, 1/500) for 30 minutes 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.
M1510-22_7.jpg Fig7: Alpha-2-macroglobulin Antibody (M1510-22) in indirect ELISA.

Indirect ELISA analysis of Alpha-2-macroglobulin was performed by coating wells of a 96-well plate with 100 µl per well of Alpha-2-macroglobulin standard diluted in carbonate/bicarbonate buffer, at a concentration of 1 µg/mL overnight at 4℃. Wells of the plate were washed, blocked with StartingBlock blocking buffer, and incubated with 100 µl per well of a Alpha-2-macroglobulin monoclonal antibody starting at a concentration of 20 µg/mL and serially diluting it to a concentration of 1.28 ng/mL for 2 hours at room temperature. The plate was washed and incubated with 100 µl per well of an HRP-conjugated goat anti-mouse IgG secondary antibody at a dilution of 1:10,000 for one hour at room temperature. Detection was performed using an Ultra TMB Substrate for 5 minutes at room temperature in the dark. The reaction was stopped with sulfuric acid and absorbances were read on a spectrophotometer at 450 nm.
Note: All products are “FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE”.