Antibody data
- Antibody Data
- Antigen structure
- References [0]
- Comments [0]
- Validations
- Western blot [3]
- Immunocytochemistry [2]
- Other assay [1]
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Validation data
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- Product number
- MA5-21203 - Provider product page
- Provider
- Invitrogen Antibodies
- Product name
- HP1 beta Monoclonal Antibody (4E12)
- Antibody type
- Monoclonal
- Antigen
- Recombinant full-length protein
- Description
- Peptide Sequence: GKKQNKKKVE EVLEEEEEEY VVEKVLDRRV VKGKVEYLLK WKGFSDEDNT WEPEENLDCP DLIAEFLQSQ KTAHETDKSE GGKRKADSDS EDKGEESK
- Reactivity
- Human, Rat
- Host
- Mouse
- Isotype
- IgG
- Antibody clone number
- 4E12
- Vial size
- 100 µg
- Concentration
- 1 mg/mL
- Storage
- -20° C, Avoid Freeze/Thaw Cycles
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Supportive validation
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Western Blot analysis of CBX1 expressed in transfected 293T cell line by anti-CBX1 monoclonal antibody (Product # MA5-21203), clone 4E12. Lane 1: CBX1 transfected lysate (21.4 KDa). Lane 2: Non-transfected lysate.
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Knockdown of HP1 was achieved by transfecting MCF7 with HP1 specific siRNAs (Silencer® select Product # s21549). Western blot analysis (Fig. a) was performed using whole cell extracts from the HP1 knockdown cells (lane 3), non-specific scrambled siRNA transfected cells (lane 2) and untransfected cells (lane 1). The blot was probed with HP1 Monoclonal Antibody (Product # MA5-21203, 1ug/ml) and Goat anti-Mouse IgG (H+L) Superclonal™ Recombinant Secondary Antibody, HRP (Product # A28177, 0.25µg/ml, 1:4000 dilution). Densitometric analysis of this western blot is shown in histogram (Fig. b). Decrease in signal upon siRNA mediated knock down confirms that antibody is specific to HP1.
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Western blot was performed using Anti-HP1 beta Monoclonal Antibody (Product # MA5-21203) and a 25kDa band corresponding to HP1 was observed across the cell lines and tissues tested. Modified whole cell extracts (30ug) of HeLa (Lane 1), MCF7 (Lane 2), A-431 (Lane 3), U-2 OS (Lane 4), PC-3 (Lane 5), Raji (Lane 6), IMR32 (Lane 7), Hep G2 (Lane 8), Mouse Liver (Lane 9) and Rat Liver (Lane 10) 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 (1ug/ml) and detected by chemiluminescence with Goat anti-Mouse IgG (H+L), Superclonal™ Recombinant Secondary Antibody, HRP (Product # A28177, 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 HeLa cells using an anti-CBX1 monoclonal antibody (Product # MA5-21203). (antibody concentration 10 µg/mL).
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Immunofluorescence analysis of HP1 was performed using MCF7 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 HP1 beta Monoclonal Antibody (Product # MA5-21203) at 5 µg/mL in 0.1% BSA and incubated overnight at 4 degree 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 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 composite image showing nuclear localization of HP1. 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
- Detection limit for recombinant GST tagged CBX1 using anti-CBX1 monoclonal antibody (Product # MA5-21203) is 0.1 ng/mL as a capture antibody.