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Western blotAntibody data
- Antibody Data
- Antigen structure
- References [1]
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- Validations
- Western blot [3]
- Immunocytochemistry [1]
- Immunohistochemistry [1]
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- Product number
- PA5-18361 - Provider product page
- Provider
- Invitrogen Antibodies
- Product name
- APOE Polyclonal Antibody
- Antibody type
- Polyclonal
- Antigen
- Synthetic peptide
- Description
- This antibody is tested in Peptide ELISA: antibody detection limit dilution 128,000.
- Reactivity
- Human
- Host
- Goat
- Isotype
- IgG
- Vial size
- 100 µg
- Concentration
- 0.5 mg/mL
- Storage
- -20° C, Avoid Freeze/Thaw Cycles
Submitted references Isolation, culture, and cryosectioning of primary porcine retinal pigment epithelium on transwell cell culture inserts.
Hood EMS, Curcio CA, Lipinski D
STAR protocols 2022 Dec 16;3(4):101758
STAR protocols 2022 Dec 16;3(4):101758
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Supportive validation
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Western blot analysis of APOE by a APOE monoclonal antibody (Product # PA5-18361) at a concentration of 2 µg/mL. Human Brain (Hippocampus) lysate (35µg protein in RIPA buffer). Detected by chemiluminescence.
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Western Blot staining of Human Brain lysate using Product # PA5-18361 at a concentration of 0.1 µg/mL, the primary antibody incubation was 1 hour and the detection method was chemiluminescence.
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Western blot was performed using Anti-APOE Polyclonal Antibody (Product # PA5-18361) and a 36 kDa band corresponding to APOE was observed in Hep G2 and Caco-2 with PTI treatment but was absent in A-431 and Daudi which are reported to be negative. Whole cell extracts (30 µg lysate) of Hep G2 (Lane 1), Hep G2 treated with Protein Transport Inhibitor (PTI; 1x for 4 hours) (Lane 2), Caco-2 (Lane 3), Caco-2 treated with Protein Transport Inhibitor (PTI; 1x for 4 hours) (Lane 4), A-431 (Lane 5), A-431 treated with Protein Transport Inhibitor (PTI; 1x for 4 hours) (Lane 6), Daudi (Lane 7) and Daudi treated with Protein Transport Inhibitor (1x for 4 hours) (Lane 8) 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 (0.1 µg/mL) and detected by chemiluminescence with Rabbit anti-Goat IgG (H+L), Superclonal™ Recombinant Secondary Antibody, HRP (Product # A27014, 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 APOE 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 APOE Goat Polyclonal Antibody (Product # PA5-18361) at 5 µg/mL in 0.1% BSA, incubated at 4 degree Celsius overnight and then labeled with Rabbit anti-Goat IgG (H+L) Cross-Adsorbed Secondary Antibody, Alexa Fluor 488 (Product # A-11078) at a dilution of 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). Panel d represents the merged image showing cytoplasmic localization. Panel e 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
- Immunohistochemical staining of paraffin embedded of Human Liver using Product # PA5-18361 at a concentration of 2.5 µg/mL. The tissue was processed by steamed antigen retrieval with citrate buffer pH 6 and stained with AP.
