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Western blot
ELISAAntibody data
- Antibody Data
- Antigen structure
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- Validations
- Western blot [1]
- Immunocytochemistry [1]
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- Product number
- ABS 038-26-02 - Provider product page
- Provider
- Invitrogen Antibodies
- Product name
- NGAL Monoclonal Antibody (26)
- Antibody type
- Monoclonal
- Antigen
- Recombinant full-length protein
- Description
- Filtration: 0.2 µm post-manufacturing filtered.
- Reactivity
- Human
- Host
- Mouse
- Isotype
- IgG
- Antibody clone number
- 26
- Vial size
- 400 µL
- Concentration
- 1 mg/mL
- Storage
- 4° C, store in dark
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Supportive validation
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Western Blot was performed using Anti-NGAL Monoclonal Antibody (26) (Product # ABS 038-26-02) and a 22 kDa band corresponding to NGAL was observed across all the tested cell lines, and was increased in HT-29 upon treatment with Thapsigargin. Whole cell extracts (30 µg lysate) of HT-29 (Lane 1), HT-29 (Thapsigargin, 500 nm, 48 hours) (Lane 2), NIH:OVCAR-3 (Lane 3), SH-SY5Y (Lane 4) were electrophoresed using NuPAGE™ 12% Bis-Tris Protein Gel (Product # NP0342BOX). Resolved proteins were then transferred onto a nitrocellulose membrane (Product # IB23002) by iBlot® 2 Dry Blotting System (Product # IB21001). The blot was probed with the primary antibody (1 µg/mL) and detected by chemiluminescence with Goat anti-Mouse IgG (H+L) Superclonal™ Recombinant Secondary Antibody, HRP (Product # A28177, 1/4000) 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 NGAL was performed using 70% confluent log phase HT-29 treated with Thapsigargin (500 nm, 48hours). 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 NGAL Monoclonal Antibody (26) (Product # ABS 038-26-02) at 5 µg/mL in 0.1% BSA, incubated at 4 degree celsius overnight and then labeled with Donkey anti-Mouse IgG (H+L) Highly Cross-Adsorbed Secondary Antibody, Alexa Fluor Plus 488 (Product # A32766, 1/2000), for 45 minutes at room temperature (Panel a: Green). Nuclei (Panel b:Blue) were stained with ProLong™ Diamond Antifade Mountant with DAPI (Product # P36962). F-actin (Panel c: Red) was stained with Rhodamine Phalloidin (Product # R415, 1:300). Panel d represents the merged image showing cytoplasmic localization. Panel e represents untreated cells showing no expression of NGAL Panel f represents control cells with no primary antibody to assess background. The images were captured at 60X magnification.
