Invitrogen

CENPA Polyclonal Antibody

Advanced Verification

This Antibody was verified by Knockdown to ensure that the antibody binds to the antigen stated. View Details

View all (19) CENPA antibodies

Datasheet

Questions & Answers

Datasheet

Questions & Answers

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CENPA Antibody in Western Blot (WB) — Knockdown of CENPA was achieved by transfecting HT-29 with CENPA specific siRNAs (Silencer® select Product # S2907, S2906). Western blot analysis (Fig. a) was performed using Nuclear enriched extracts from the CENPA knockdown cells (lane 3), non-targeting scrambled siRNA transfected cells (lane 2) and untransfected cells (lane 1). The blot was probed with CENPA Polyclonal Antibody (Product # PA5-17194, 1:1000 ) and Goat anti-Rabbit IgG (H+L) Superclonal™ Recombinant Secondary Antibody, HRP (Product # A27036, 1:4000). 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 CENPA.

CENPA Antibody in Immunocytochemistry (ICC) — Immunofluorescence analysis of CENPA was performed using 70% confluent log phase HT-29 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 45 minutes at room temperature. The cells were labeled with CENPA Polyclonal Antibody (Product # PA5-17194) 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), (1:2000), 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 Centromere localization. Panel e represents control cells with no primary antibody to assess background. The images were captured at 60X magnification.

CENPA Antibody in Knockdown — Antibody specificity was demonstrated by siRNA mediated knockdown of target protein. HT-29 cells were transfected with CENPA siRNA and decrease in signal intensity was observed in Western Blot application using Anti-CENPA Polyclonal Antibody (Product # PA5-17194). {KD}

Product Details

Applications

Tested Dilution

Publications

Western Blot (WB)

1:1000

Immunocytochemistry (ICC)

1:200

Immunofluorescence (IF)

1:200

Product Specifications

Species Reactivity

Human, Rat

Host/Isotype

Rabbit / IgG

Class

Polyclonal

Type

Antibody

Immunogen

Synthetic peptide corresponding to human CENP-A protein

Conjugate

Unconjugated

Form

Liquid

Purification

Antigen affinity chromatography

Storage buffer

0.01M HEPES, pH 7.5, with 0.15M NaCl, 100µg/mL BSA, 50% glycerol

Contains

no preservative

Storage conditions

-20°C

RRID

AB_10987425

Product Specific Information

It is not recommended to aliquot this antibody.

This antibody is not cross-reactive with other histone proteins, including Histone H3.

Target Information

Centromeres are the differentiated chromosomal domains that specify the mitotic behavior of chromosomes. CENPA encodes a centromere protein which contains a histone H3 related histone fold domain that is required for targeting to the centromere. CENPA is proposed to be a component of a modified nucleosome or nucleosome-like structure in which it replaces 1 or both copies of conventional histone H3 in the (H3-H4)2 tetrameric core of the nucleosome particle.

For Research Use Only. Not for use in diagnostic procedures. Not for resale without express authorization.

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Bioinformatics

Protein Aliases: CENP A; CENP-A; Centromere autoantigen A; Centromere protein A; centromere protein A, 17kDa; centromere-specific histone; Histone H3-like centromeric protein A

Gene Aliases: CenH3; CENP-A; CENPA; RGD1563607

UniProt ID: (Human) P49450

Entrez Gene ID: (Human) 1058, (Rat) 298850

Molecular Function: DNA binding protein histone nucleic acid binding