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Cbx8 Recombinant Rabbit Monoclonal Antibody [PSH06-61]

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Specification

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Catalog# HA722734

Cbx8 Recombinant Rabbit Monoclonal Antibody [PSH06-61]

Application

  • WB

  • IF-Cell

  • FC

  • IP

Reactivity

  • Human

  • Mouse

  • Rat

  • Green monkey

BSA and Azide free
Conjugation

  • unconjugated

This product has been cited in peer reviewed publications, see list HERE

Overview

Product Name

Cbx8 Recombinant Rabbit Monoclonal Antibody [PSH06-61]

Antibody Type

Recombinant Rabbit monoclonal Antibody

Immunogen

Recombinant protein within human Cbx8 aa 51-389.

Species Reactivity

Human, Mouse, Rat, Green monkey

Validated Applications

WB, IF-Cell, FC, IP

Molecular Weight

Predicted band size: 43 kDa

Positive Control

HCT 116 cell lysate, HeLa cell lysate, SW620 cell lysate, U-2 OS cell lysate, NIH/3T3 cell lysate, SP2/0 cell lysate, PC-12 cell lysate, C6 cell lysate, COS-1 cell lysate, HeLa, NIH/3T3.

Conjugation

unconjugated

Clone Number

PSH06-61

Product Features

Form

Liquid

Concentration

Storage Instructions

Shipped at 4℃. Store at +4℃ short term (1-2 weeks). Store at -20℃ long term.

Storage Buffer

PBS (pH7.4), 0.1% BSA, 40% Glycerol. Preservative: 0.05% Sodium Azide.

Isotype

IgG

Purification Method

Protein A affinity purified.

Application Dilution

  • WB

  • 1:1,000

  • IF-Cell

  • 1:100

  • FC

  • 1:1,000

  • IP

  • 1-2μg/sample

Target

Function

Component of a Polycomb group (PcG) multiprotein PRC1-like complex, a complex class required to maintain the transcriptionally repressive state of many genes, including Hox genes, throughout development. PcG PRC1 complex acts via chromatin remodeling and modification of histones; it mediates monoubiquitination of histone H2A 'Lys-119', rendering chromatin heritably changed in its expressibility. Enables methylated histone binding activity. Involved in negative regulation of transcription by RNA polymerase II. Located in chromatin and nucleoplasm. Part of PRC1 complex. Biomarker of esophagus squamous cell carcinoma and glioblastoma.

Background References

1. Chen H et al. CBX8 promotes lung adenocarcinoma growth and metastasis through transcriptional repression of CDKN2C and SCEL. J Cell Physiol. 2023 Nov

2. Suh JL et al. Reprogramming CBX8-PRC1 function with a positive allosteric modulator. Cell Chem Biol. 2022 Apr

Subcellular Location

Nucleus.

UNIPROT

SwissProt: Q9HC52 Human

SwissProt: Q9QXV1 Mouse

Entrez Gene: 303731 Rat

Synonyms

Cbx8 antibody

CBX8_HUMAN antibody

Chromobox Homolog 8 (Drosophila Pc Class) antibody

Chromobox Homolog 8 (Pc Class Homolog, Drosophila) antibody

Chromobox Homolog 8 antibody

Chromobox protein homolog 8 antibody

hPc3 antibody

Pc Class 3 Homolog (Drosophila) antibody

Pc Class 3 Homolog antibody

Pc3 antibody

Expand

Images

  • Western blot analysis of Cbx8 on different lysates with Rabbit anti-Cbx8 antibody (<a href="/products/HA722734" style="font-weight: bold;text-decoration: underline;">HA722734</a>) at 1/1,000 dilution.<br /><br />Lane 1: HCT 116 cell lysate<br />Lane 2: HeLa cell lysate<br />Lane 3: SW620 cell lysate<br />Lane 4: U-2 OS cell lysate<br />Lane 5: NIH/3T3 cell lysate<br />Lane 6: SP2/0 cell lysate<br />Lane 7: PC-12 cell lysate<br />Lane 8: C6 cell lysate<br />Lane 9: COS-1 cell lysate<br /><br />Lysates/proteins at 20 µg/Lane.<br /><br />Predicted band size: 43/40 kDa<br />Observed band size: 43/40 kDa<br /><br />Exposure time: 20 seconds; ECL: K1801;<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/HA722734" style="font-weight: bold;text-decoration: underline;">HA722734</a>) at 1/1,000 dilution was used in 5% NFDM/TBST at 4℃ overnight. Goat Anti-Rabbit IgG - HRP Secondary Antibody (<a href="/products/HA1001" style="font-weight: bold;text-decoration: underline;">HA1001</a>) at 1/50,000 dilution was used for 1 hour at room temperature.

    Western blot analysis of Cbx8 on different lysates with Rabbit anti-Cbx8 antibody (HA722734) at 1/1,000 dilution.

    Lane 1: HCT 116 cell lysate
    Lane 2: HeLa cell lysate
    Lane 3: SW620 cell lysate
    Lane 4: U-2 OS cell lysate
    Lane 5: NIH/3T3 cell lysate
    Lane 6: SP2/0 cell lysate
    Lane 7: PC-12 cell lysate
    Lane 8: C6 cell lysate
    Lane 9: COS-1 cell lysate

    Lysates/proteins at 20 µg/Lane.

    Predicted band size: 43/40 kDa
    Observed band size: 43/40 kDa

    Exposure time: 20 seconds; ECL: K1801;

    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 (HA722734) at 1/1,000 dilution was used in 5% NFDM/TBST at 4℃ overnight. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1/50,000 dilution was used for 1 hour at room temperature.

  • Immunocytochemistry analysis of HeLa cells labeling Cbx8 with Rabbit anti-Cbx8 antibody (<a href="/products/HA722734" style="font-weight: bold;text-decoration: underline;">HA722734</a>) at 1/100 dilution.<br /><br />Cells were fixed in 4% paraformaldehyde for 20 minutes at room temperature, permeabilized with 0.1% Triton X-100 in PBS for 5 minutes at room temperature, then blocked with 1% BSA in 10% negative goat serum for 1 hour at room temperature. Cells were then incubated with Rabbit anti-Cbx8 antibody (<a href="/products/HA722734" style="font-weight: bold;text-decoration: underline;">HA722734</a>) at 1/100 dilution in 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 Cbx8 with Rabbit anti-Cbx8 antibody (HA722734) at 1/100 dilution.

    Cells were fixed in 4% paraformaldehyde for 20 minutes at room temperature, permeabilized with 0.1% Triton X-100 in PBS for 5 minutes at room temperature, then blocked with 1% BSA in 10% negative goat serum for 1 hour at room temperature. Cells were then incubated with Rabbit anti-Cbx8 antibody (HA722734) at 1/100 dilution in 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.

  • Immunocytochemistry analysis of NIH/3T3 cells labeling Cbx8 with Rabbit anti-Cbx8 antibody (<a href="/products/HA722734" style="font-weight: bold;text-decoration: underline;">HA722734</a>) at 1/100 dilution.<br /><br />Cells were fixed in 4% paraformaldehyde for 20 minutes at room temperature, permeabilized with 0.1% Triton X-100 in PBS for 5 minutes at room temperature, then blocked with 1% BSA in 10% negative goat serum for 1 hour at room temperature. Cells were then incubated with Rabbit anti-Cbx8 antibody (<a href="/products/HA722734" style="font-weight: bold;text-decoration: underline;">HA722734</a>) at 1/100 dilution in 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 NIH/3T3 cells labeling Cbx8 with Rabbit anti-Cbx8 antibody (HA722734) at 1/100 dilution.

    Cells were fixed in 4% paraformaldehyde for 20 minutes at room temperature, permeabilized with 0.1% Triton X-100 in PBS for 5 minutes at room temperature, then blocked with 1% BSA in 10% negative goat serum for 1 hour at room temperature. Cells were then incubated with Rabbit anti-Cbx8 antibody (HA722734) at 1/100 dilution in 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.

  • Flow cytometric analysis of HeLa cells labeling Cbx8.<br /><br />Cells were fixed and permeabilized. Then stained with the primary antibody (<a href="/products/HA722734" style="font-weight: bold;text-decoration: underline;">HA722734</a>, 1/1,000) (red) compared with Rabbit IgG Isotype Control (green). After incubation of the primary antibody at +4℃ for an hour, the cells were stained with a iFluor&trade; 488 conjugate-Goat anti-Rabbit IgG Secondary antibody (<a href="/products/HA1121" style="font-weight: bold;text-decoration: underline;">HA1121</a>) at 1/1,000 dilution for 30 minutes at +4℃. Unlabelled sample was used as a control (cells without incubation with primary antibody; black).

    Flow cytometric analysis of HeLa cells labeling Cbx8.

    Cells were fixed and permeabilized. Then stained with the primary antibody (HA722734, 1/1,000) (red) compared with Rabbit IgG Isotype Control (green). After incubation of the primary antibody at +4℃ for an hour, the cells were stained with a iFluor™ 488 conjugate-Goat anti-Rabbit IgG Secondary antibody (HA1121) at 1/1,000 dilution for 30 minutes at +4℃. Unlabelled sample was used as a control (cells without incubation with primary antibody; black).

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Citation