Antibody data
- Antibody Data
- Antigen structure
- References [1]
- Comments [0]
- Validations
- Western blot [1]
- Immunocytochemistry [1]
- Other assay [1]
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Validation data
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- Product number
- PA1-40308 - Provider product page
- Provider
- Invitrogen Antibodies
- Product name
- Lysozyme Polyclonal Antibody
- Antibody type
- Polyclonal
- Antigen
- Other
- Description
- PA1-40308 detects Lysozyme from human samples.
- Concentration
- 0.1 mg/mL
Submitted references ATG4B/autophagin-1 regulates intestinal homeostasis and protects mice from experimental colitis.
Cabrera S, Fernández AF, Mariño G, Aguirre A, Suárez MF, Español Y, Vega JA, Laurà R, Fueyo A, Fernández-García MS, Freije JM, Kroemer G, López-Otín C
Autophagy 2013 Aug;9(8):1188-200
Autophagy 2013 Aug;9(8):1188-200
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Supportive validation
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Western blot was performed using Anti-Lysozyme Polyclonal Antibody (Product # PA1-40308) and a 15kDa band corresponding to Lysozyme was observed across cell lines tested except HEL 92.1.7, SK-O-V3, SK-BR-3. An uncharacterized band (*) was also observed at ~30kDa. Whole cell extracts (30 µg lysate) of Hep G2 (Lane 1), THP-1 (Lane 2), HL-60 (Lane 3), HEL 92.1.7 (Lane 4), SK-O-V3 (Lane 5) and SK-BR-3 (Lane 6) were electrophoresed using Novex® NuPAGE® 4-12 % Bis-Tris gel (Product # NP0322BOX). Resolved proteins were then transferred onto a nitrocellulose membrane (Product # IB23001) by iBlot® 2 Dry Blotting System (Product # IB21001). The blot was probed with the primary antibody (1:100 dilution) and detected by chemiluminescence with Goat anti-Rabbit IgG (H+L), Superclonal™ Recombinant Secondary Antibody, HRP (Product # A27036, 1:4000 dilution) using the iBright FL 1000 (Product # A32752). Chemiluminescent detection was performed using Novex® ECL Chemiluminescent Substrate Reagent Kit (Product # WP20005).
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 Rabbit Polyclonal Antibody (Product # PA1-40308) at 1:100 dilution in 0.1% BSA, incubated at 4 degree Celsius overnight and then labeled with Goat anti-Rabbit IgG (H+L) Superclonal™ Recombinant Secondary Antibody, Alexa Fluor® 488 conjugate (Product # A27034) 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.
Supportive validation
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- NULL