MA1-83313

antibody from Invitrogen Antibodies

Targeting: LYZ

Immunocytochemistry (Supportive data in Antibodypedia)

Other assay (Supportive data in Antibodypedia)

Antibody Data

Product number

MA1-83313

Provider

Invitrogen Antibodies

Product name

Lysozyme Monoclonal Antibody (SB1)

Antibody type

Monoclonal

Antigen

Purifed from natural sources

Description

This antibody does not cross-react with lysozyme from chicken egg white.

Antibody clone number

SB1

Concentration

1 mg/mL

References

Submitted references

Lysozyme and bilirubin bind to ACE and regulate its conformation and shedding.

Danilov SM, Lünsdorf H, Akinbi HT, Nesterovitch AB, Epshtein Y, Letsiou E, Kryukova OV, Piegeler T, Golukhova EZ, Schwartz DE, Dull RO, Minshall RD, Kost OA, Garcia JG
Scientific reports 2016 Oct 13;6:34913

Immunocytochemistry

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Immunofluorescence analysis of Lysozyme was performed using 70% confluent log phase Hep G2 cells. The cells were fixed with 4% paraformaldehyde for 10 minutes, permeabilized with 0.1% Triton™ X-100 for 15 minutes, and blocked with 2% BSA for 1 hour at room temperature. The cells were labeled with Lysozyme Monoclonal Antibody (SB1) (Product # MA1-83313) at 1:100 dilution in 0.1% BSA, incubated at 4 degree Celsius overnight and then labeled with Goat anti-Mouse IgG (H+L), Superclonal™ Recombinant Secondary Antibody, Alexa Fluor 488 conjugate (Product # A28175) at a dilution of 1:2000 for 45 minutes at room temperature (Panel a: green). Nuclei (Panel b: blue) were stained with SlowFade® Gold Antifade Mountant with DAPI (Product # S36938). F-actin (Panel c: red) was stained with Rhodamine Phalloidin (Product # R415, 1:300). Panel d represents the merged image showing localization to golgi network and cytoplasm. Panel e shows SK-BR-3 cells with no expression of Lysozyme. Panel f represents control cells with no primary antibody to assess background. The images were captured at 60X magnification.

Other assay

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Figure 5 Effect of lysozyme expression on ACE shedding. CHO - ACE cells were transiently transfected with plasmids coding for human lysozyme (HLZ) or for surfactant protein C (SP-C as negative control). After 48 hours, ACE activity and lysozyme expression were quantified in the cell lysates and in the culture medium. ( A ) Western blotting of lysate and culture medium with anti-human lysozyme Ab. Positive control for the presence of lysozyme - human bronchoalveolar lavage fluid (H. BALF). ( B ) Lysozyme muramidase activity in the same samples was determined using killed Micrococcus lysodeiticus . ( C ) Soluble and cell-bound ACE activity was determined with Z-Phe-His-Leu as a substrate. The rate of ACE shedding was calculated as ACE activity in the culture medium/the total amount of cell-associated and secreted ACE activity. Data were expressed as a percentage from negative control. Mean + SD of three experiments.

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Figure 4 Identification of lysozyme as ACE-associated protein. CHO cells expressing testicular (CHO-tACE) and somatic (CHO-ACE) isoforms of ACE were pre-incubated overnight with human lysozyme or BSA (as negative control) at 250 mug/ml. Lysates of these cells were precipitated with anti-ACE and anti-lysozyme antibodies. Representative Western blots showed co-precipitation of ACE and lysozyme from CHO-ACE cells. Identical results were obtained in two additional experiments.