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CENPA Recombinant Rabbit Monoclonal Antibody [PSH0-09] - BSA and Azide free

Usd: 649

Specification

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Catalog# HA750578

CENPA Recombinant Rabbit Monoclonal Antibody [PSH0-09] - BSA and Azide free

Application

  • WB

  • IF-Cell

Reactivity

  • Human

With BSA and Azide
Conjugation

  • unconjugated

Overview

Product Name

CENPA Recombinant Rabbit Monoclonal Antibody [PSH0-09] - BSA and Azide free

Antibody Type

Recombinant Rabbit monoclonal Antibody

Immunogen

Synthetic peptide within human CENPA aa 21-70 / 140.

Species Reactivity

Human

Validated Applications

WB, IF-Cell

Molecular Weight

Predicted band size: 16 kDa

Positive Control

Hela cell lysate, A375 cell lysate

Conjugation

unconjugated

Clone Number

PSH0-09

Product Features

Form

Liquid

Concentration

1mg/ml

Storage Instructions

Store at +4℃ after thawing. Aliquot store at -20℃. Avoid repeated freeze / thaw cycles.

Storage Buffer

1*PBS (pH7.4).

Isotype

IgG

Purification Method

Protein A affinity purified.

Application Dilution

  • WB

  • 1:1,000

  • IF-Cell

  • 1:100

Target

Function

Centromere protein A, also known as CENPA, is a protein which in humans is encoded by the CENPA gene. CENPA is a histone H3 variant which is the critical factor determining the kinetochore position(s) on each chromosome in most eukaryotes including humans. CENPA is a protein which epigenetically defines the position of the centromere on each chromosome, determining the position of kinetochore assembly and the final site of sister chromatid cohesion during mitosis. The CENPA protein is a histone H3 variant which replaces one or both canonical H3 histones in a subset of nucleosomes within centromeric chromatin. CENPA has the greatest sequence divergence of the histone H3 variants, with just 48% similarity to canonical histone H3, and has a highly diverged N-terminal tail that lacks many well characterised histone modification sites including H3K4, H3K9 and H3K27. Unusually for a histone, CENPA nucleosomes are not loaded together with DNA replication and are loaded at different cell cycle stages in different organisms: G1 phase in human, M phase in drosophila, G2 in S. pombe. To orchestrate this specialised loading there are CENPA-specific histone chaperones: HJURP in human, CAL1 in drosophila and Scm3 in S. pombe. In most eukaryotes CENPA is loaded into large domains of highly repetitive satellite DNA. The position of CENPA within satellite DNA are heritable at the protein level through a purely epigenetic mechanism. This means that the position of CENPA protein binding to the genome is copied upon cell division to the two daughter cells independent of the underlying DNA sequence. Under circumstances in which CENPA is lost from a chromosome a fail-safe mechanism has been described in human cells in which CENPB recruits CENPA via a satellite DNA binding domain to repopulate the centromere with CENPA nucleosomes. CENPA interacts directly with the inner kinetochore through proteins including CENPC and CENPN. Through this interaction the microtubules are able to accurately segregate chromosomes during mitosis.

Background References

1. Wang Q et al. CENPA promotes clear cell renal cell carcinoma progression and metastasis via Wnt/β-catenin signaling pathway. J Transl Med. 2021 Oct

2. Han J et al. CENPA is one of the potential key genes associated with the proliferation and prognosis of ovarian cancer based on integrated bioinformatics analysis and regulated by MYBL2. Transl Cancer Res. 2021 Sep

Subcellular Location

Nucleus, Chromosome, centromere, kinetochore.

UNIPROT

SwissProt: P49450 Human

Synonyms

CENP A antibody

CENP-A antibody

cenpa antibody

CENPA_HUMAN antibody

Centromere autoantigen A antibody

Centromere protein A 17kDa antibody

Centromere protein A antibody

Histone H3 like centromeric protein A antibody

Histone H3-like centromeric protein A antibody

Images

  • Western blot analysis of CENPA on different lysates with Rabbit anti-CENPA antibody (<a href="/products/HA750578" style="font-weight: bold;text-decoration: underline;">HA750578</a>) at 1/1,000 dilution.<br /><br />Lane 1: Hela cell lysate<br />Lane 2: A375 cell lysate<br /><br />Lysates/proteins at 10 µg/Lane.<br /><br />Predicted band size: 16 kDa<br />Observed band size: 16 kDa<br /><br />Exposure time: 5 minutes;<br /><br />4-20% SDS-PAGE gel.<br /><br />Proteins were transferred to a PVDF membrane and blocked with 5% NFDM/TBST for 1 hour at room temperature. The primary antibody (<a href="/products/HA750578" style="font-weight: bold;text-decoration: underline;">HA750578</a>) at 1/1,000 dilution was used in 5% NFDM/TBST at room temperature for 2 hours. Goat Anti-Rabbit IgG - HRP Secondary Antibody (<a href="/products/HA1001" style="font-weight: bold;text-decoration: underline;">HA1001</a>) at 1:300,000 dilution was used for 1 hour at room temperature.

    Western blot analysis of CENPA on different lysates with Rabbit anti-CENPA antibody (HA750578) at 1/1,000 dilution.

    Lane 1: Hela cell lysate
    Lane 2: A375 cell lysate

    Lysates/proteins at 10 µg/Lane.

    Predicted band size: 16 kDa
    Observed band size: 16 kDa

    Exposure time: 5 minutes;

    4-20% SDS-PAGE gel.

    Proteins were transferred to a PVDF membrane and blocked with 5% NFDM/TBST for 1 hour at room temperature. The primary antibody (HA750578) at 1/1,000 dilution was used in 5% NFDM/TBST at room temperature for 2 hours. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1:300,000 dilution was used for 1 hour at room temperature.

  • Immunocytochemistry analysis of HeLa cells labeling CENPA with Rabbit anti-CENPA antibody (<a href="/products/HA750578" style="font-weight: bold;text-decoration: underline;">HA750578</a>) at 1/100 dilution.<br /><br />Cells were fixed in 4% paraformaldehyde for 10 minutes at room temperature, permeabilized with 0.1% Triton X-100 in PBS for 15 minutes at room temperature, then blocked with 2% BSA in 10% negative goat serum for 1 hour at room temperature. Cells were then incubated with Rabbit anti-CENPA antibody (<a href="/products/HA750578" style="font-weight: bold;text-decoration: underline;">HA750578</a>) at 1/100 dilution in 0.1% BSA in PBST overnight at 4 ℃. Goat Anti-Rabbit IgG H&L (iFluor&trade; 488, <a href="/products/HA1121" style="font-weight: bold;text-decoration: underline;">HA1121</a>) was used as the secondary antibody at 1/1,000 dilution. PBS instead of the primary antibody was used as the secondary antibody only control. Nuclear DNA was labelled in blue with DAPI.<br /><br />Beta tubulin (<a href="/products/M1305-2" style="font-weight: bold;text-decoration: underline;">M1305-2</a>, red) was stained at 1/100 dilution overnight at +4℃. Goat Anti-Mouse IgG H&L (iFluor&trade; 594, <a href="/products/HA1126" style="font-weight: bold;text-decoration: underline;">HA1126</a>) was used as the secondary antibody at 1/1,000 dilution.

    Immunocytochemistry analysis of HeLa cells labeling CENPA with Rabbit anti-CENPA antibody (HA750578) at 1/100 dilution.

    Cells were fixed in 4% paraformaldehyde for 10 minutes at room temperature, permeabilized with 0.1% Triton X-100 in PBS for 15 minutes at room temperature, then blocked with 2% BSA in 10% negative goat serum for 1 hour at room temperature. Cells were then incubated with Rabbit anti-CENPA antibody (HA750578) at 1/100 dilution in 0.1% BSA in PBST overnight at 4 ℃. Goat Anti-Rabbit IgG H&L (iFluor™ 488, HA1121) was used as the secondary antibody at 1/1,000 dilution. PBS instead of the primary antibody was used as the secondary antibody only control. Nuclear DNA was labelled in blue with DAPI.

    Beta tubulin (M1305-2, red) was stained at 1/100 dilution overnight at +4℃. Goat Anti-Mouse IgG H&L (iFluor™ 594, HA1126) was used as the secondary antibody at 1/1,000 dilution.

Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"