Antibody data
- Antibody Data
- Antigen structure
- References [0]
- Comments [0]
- Validations
- Western blot [2]
- Immunocytochemistry [2]
- Chromatin Immunoprecipitation [1]
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Validation data
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- Product number
- PA5-65869 - Provider product page
- Provider
- Invitrogen Antibodies
- Product name
- CENPB Polyclonal Antibody
- Antibody type
- Polyclonal
- Antigen
- Recombinant full-length protein
- Description
- Immunogen sequence: TFREKSRIIQE VEENPDLRKG EIARRFNIPP STLSTILKNK RAILASERKY GVASTCRKTN KLSPYDKLEG LLIAWFQQIR AAGLPVKGII L Highest antigen sequence identity to the following orthologs - mouse 100%, rat 100%.
- Reactivity
- Human
- Host
- Rabbit
- Isotype
- IgG
- Vial size
- 100 µL
- Concentration
- 0.08 mg/mL
- Storage
- Store at 4°C short term. For long term storage, store at -20°C, avoiding freeze/thaw cycles.
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Supportive validation
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- KD of CENPB was achieved by transfecting PC-3 with CENPB specific siRNAs (Silencer® select Product # s2909, s2910). Western blot analysis (Fig. a) was performed using modified whole cell extracts (1% SDS) from the CENPB KD cells (Lane 3), non-specific scrambled siRNA transfected cells (Lane 2) and untransfected cells (Lane 1). The blot was probed with CENPB Polyclonal Antibody (Product # PA5-65869, 1:500 dilution) and Goat anti-Rabbit IgG (H+L) Superclonal™ Secondary Antibody, HRP conjugate (Product # A27036, 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 CENPB.
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Western blot was performed using Anti-CENPB Polyclonal Antibody (Product # PA5-65869) and 80 kDa band corresponding to CENPB was observed across cell lines tested. Modified whole cell extracts (1% SDS) (30 µg lysate) of PC-3 (Lane 1), MCF7 (Lane 2), A549 (Lane 3), A-431 (Lane 4), HeLa (Lane 5), Jurkat (Lane 6) and KARPAS 299 (Lane 7) 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 (1:500 dilution) and detected by Goat Anti-Rabbit IgG Secondary Antibody, HRP conjugate (Product # A27036, 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
- Immunofluorescent staining of CENPB in human cell line A-431 shows localization to nuclear bodies. Samples were probed using a CENPB Polyclonal Antibody (Product # PA5-65869).
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Immunofluorescence analysis of CENPB was performed using 70% confluent log phase PC-3 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 CENPB Polyclonal Antibody (Product # PA5-65869) at 5 microgram/mL in 0.1% BSA, incubated at 4 degree Celsius overnight and then labeled with Goat anti-Rabbit IgG (H+L) Superclonal™ 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 merged image showing localization to nuclear bodies. Panel e represents 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
- Enrichment of endogenous CENPB protein at specific gene loci using Anti-CENPB Antibody: Chromatin Immunoprecipitation (ChIP) was performed using Anti-CENPB Rabbit Polyclonal Antibody (Product # PA5-65869, 5 µg) on sheared chromatin from 2 million Hela cells using the MAGnify ChIP System kit (Product # 49-2024). Normal Rabbit IgG was used as a negative IP control. The purified DNA was analyzed by qPCR with PCR primer pairs over SATa-Chr3 and SATa-Chr5. Data is presented as fold enrichment of the antibody signal versus the negative control IgG using the comparative CT method.