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Western blotAntibody data
- Antibody Data
- Antigen structure
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- Validations
- Western blot [3]
- Immunocytochemistry [1]
- Immunohistochemistry [1]
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- Product number
- MA5-33052 - Provider product page
- Provider
- Invitrogen Antibodies
- Product name
- Chromogranin A Recombinant Rabbit Monoclonal Antibody (RM385)
- Antibody type
- Monoclonal
- Antigen
- Synthetic peptide
- Description
- Specificity: reacts to Human Chromogranin A.
- Antibody clone number
- RM385
- Concentration
- 0.1 mg/mL
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Supportive validation
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Western blot analysis of Chromogranin A in human brain tissue lysate. Samples were incubated with Chromogranin A monoclonal antibody (Product # MA5-33052) using a 1:1000 dilution.
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Western blot analysis of Chromogranin A in human brain tissue lysate. Samples were incubated with Chromogranin A monoclonal antibody (Product # MA5-33052) using a 1:1000 dilution.
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Western blot was performed using Anti-Chromogranin A Recombinant Rabbit Monoclonal Antibody (RM385) (Product # MA5-33052) and a 65 kDa band corresponding to Chga was observed across cell lines along with an uncharacterized band (*). Whole cell extracts (30 µg lysate) of SH-SY5Y (Lane 1), SH-SY5Y treated with PTI (1X for 4hrs) (Lane 2), Hep G2 (Lane 3), and Hep G2 treated with PTI (1X for 4hrs) (Lane 4) were electrophoresed using NuPAGE™ 4-12% Bis-Tris Protein Gel (Product # NP0322BOX). 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 (1:1000) and detected by chemiluminescence with Goat anti-Rabbit IgG (H+L) Superclonal™ Recombinant Secondary Antibody, HRP (Product # A27036,1:20000) using the iBright™ FL1500 Imaging System (Product # A44115). Chemiluminescent detection was performed using SuperSignal™ West Pico PLUS Chemiluminescent Substrate (Product # 34580).
Supportive validation
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Immunofluorescence analysis of Chga was performed using 70% confluent log phase SH-SY5Y treated with PTI (1X for 4hrs) 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 Chromogranin A Recombinant Rabbit Monoclonal Antibody (RM385) (Product # MA5-33052) at 1:100 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), (1:2000), 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 Cytoplasm localization. Panel e represents HepG2 cells treated with PTI showing low expression of Chga. 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
- Immunohistochemistry analysis of Chromogranin A in paraffin-embedded human colon cancer tissue section. Samples were incubated with Chromogranin A monoclonal antibody (Product # MA5-33052) using a 1:10,000 dilution.
