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Western blotAntibody data
- Antibody Data
- Antigen structure
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- Validations
- Western blot [2]
- Immunocytochemistry [1]
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- Product number
- PA5-21410 - Provider product page
- Provider
- Invitrogen Antibodies
- Product name
- HLA-DR beta Polyclonal Antibody
- Antibody type
- Polyclonal
- Antigen
- Recombinant protein fragment
- Description
- Recommended positive controls: Raji.
- Concentration
- 1 mg/mL
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Supportive validation
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Western Blot using HLA-DR beta Polyclonal Antibody (Product # PA5-21410). Sample (30 µg whole cell lysate). A: Raji. 12% SDS PAGE. HLA-DR beta Polyclonal Antibody (Product # PA5-21410) diluted at 1:1,000.
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Western blot was performed using Anti-HLA-DR beta Polyclonal Antibody (Product # PA5-21410) and a 30k Da band corresponding to HLA-DR beta was observed in THP-1 cells, THP-1 differentiated into macrophages and dendritic cells, Raji and Ramos but not in Jurkat and MOLT-4. Membrane enriched extracts (30 µg lysate) of THP-1 (Lane 1), THP-1 differentiated macrophages (Lane 2), THP-1 differentiated dendritic cells(Lane 3), Raji (Lane 4), Ramos (Lane 5), Jurkat (Lane 6) or MOLT-4 (Lane 7) were electrophoresed using NuPAGE™ 10% Bis-Tris Protein Gel (Product # NP0302BOX). 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 (1:1000) and detected by chemiluminescence with Goat anti-Rabbit IgG (H+L) Superclonal™ Recombinant Secondary Antibody, HRP (Product # A27036,1:4000) 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 HLA-DR beta was performed using 70% confluent log phase Raji cells. The cells were fixed with 4% paraformaldehyde for 10 minutes, permeabilized with 0.1% Triton™ X-100 for 10 minutes, and blocked with 2% BSA for 45 minutes at room temperature. The cells were labeled with HLA-DR beta Polyclonal Antibody (Product # PA5-21410) at 1:200 in 0.1% BSA, incubated at 4 degree celsius overnight and then labeled with Donkey anti-Rabbit IgG (H+L) Highly Cross-Adsorbed Secondary Antibody, Alexa Fluor Plus 488 (Product # A32790), (1:2500), 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 plasma membrane localization. Panel e shows no expression in Jurkat cells. Panel f represents control cells with no primary antibody to assess background. The images were captured at 63X magnification.
