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Phospho-Histone H3.3 (S31) Recombinant Rabbit Monoclonal Antibody [JE62-92]

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Specification

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Catalog# HA721034

Phospho-Histone H3.3 (S31) Recombinant Rabbit Monoclonal Antibody [JE62-92]

Application

  • WB

  • IF-Cell

Reactivity

  • Human

  • Rat

Conjugation

  • unconjugated

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

Overview

Product Name

Phospho-Histone H3.3 (S31) Recombinant Rabbit Monoclonal Antibody [JE62-92]

Antibody Type

Recombinant Rabbit monoclonal Antibody

Immunogen

Synthetic phospho-peptide corresponding to residues surrounding Ser31 of Human Histone H3.3.

Species Reactivity

Human, Rat

Validated Applications

WB, IF-Cell

Target Molecular Weight

Predicted band size: 15 kDa

Positive Control

HeLa treated with 100ng/mL Nocodazole for 18 hours cell lysate, PC-12 treated with 100ng/mL Nocodazole for 18 hours cell lysate, HeLa cells treated with 100ng/mL Nocodazole for 18 hours.

Conjugation

unconjugated

Clone Number

JE62-92

RRID

AB_3072158

Product Features

Form

Liquid

Concentration

1 mg/mL.(The concentration of this product may be batch-dependent)

Storage Instructions

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

Storage Buffer

1*TBS (pH7.4), 0.05% BSA, 40% Glycerol. Preservative: 0.05% Sodium Azide.

Isotype

IgG

Purification Method

Protein A affinity purified.

Application Dilution

  • WB

  • 1:500-1:1,000

  • IF-Cell

  • 1:25,000

Target

Function

Variant histone H3 which replaces conventional H3 in a wide range of nucleosomes in active genes. Constitutes the predominant form of histone H3 in non-dividing cells and is incorporated into chromatin independently of DNA synthesis. Deposited at sites of nucleosomal displacement throughout transcribed genes, suggesting that it represents an epigenetic imprint of transcriptionally active chromatin. Nucleosomes wrap and compact DNA into chromatin, limiting DNA accessibility to the cellular machineries which require DNA as a template. Histones thereby play a central role in transcription regulation, DNA repair, DNA replication and chromosomal stability. DNA accessibility is regulated via a complex set of post-translational modifications of histones, also called histone code, and nucleosome remodeling.

Background References

1. Strobino M. et. al. Loss of histone H3.3 results in DNA replication defects and altered origin dynamics in C. elegans. Genome Res. 2020 Dec

2. Ince AT. et. al. The relationship of Serum Histone H3.3 and H4 with chronic Hepatitis B. J Pak Med Assoc. 2020 Sep

Subcellular Location

Nucleus, Chromosome.

UNIPROT

SwissProt: P84243 Human

SwissProt: P84245 Rat

Synonyms

H3 histone family 3A antibody

H3 histone family 3B antibody

H3 histone, family 3B (H3.3B) antibody

H3.3 antibody

H3.3A antibody

H3.3B antibody

H33_HUMAN antibody

H3F3 antibody

H3F3A antibody

H3f3b antibody

Expand

Images

  • <span style="font-weight: bold;">☑ Cell treatment (CT)</span><br /><br />Western blot analysis of Histone H3.3 (phospho S31) on Hela cell lysates.<br /><br />Lane 1: Hela cells, whole cell lysate, 10ug/lane<br />Lane 2/3: Hela cells treated with 100ng/ml Nocodazole for 18h, whole cell lysates, 10ug/lane<br />Lane 4: Hela cells treated with 100ng/ml Nocodazole for 18h, then treated with 2.8ug/ul lambda-PP for 30 minutes, whole cell lysates, 10ug/lane<br /><br />All lanes :<br />Anti-Histone H3.3 (phospho S31) antibody (<a href="/products/HA721034" style="font-weight: bold;text-decoration: underline;">HA721034</a>) at 1:500 dilution. Anti-GAPDH antibody (<a href="/products/ET1601-4" style="font-weight: bold;text-decoration: underline;">ET1601-4</a>) at 1:10,000 dilution. Goat Anti-Rabbit IgG H&L (HRP) (<a href="/products/HA1001" style="font-weight: bold;text-decoration: underline;">HA1001</a>) at 1/200,000 dilution.<br /><br />Predicted band size: 15 kDa<br />Observed band size: 15 kDa<br /><br />Blocking and diluting buffer: 5% BSA.<br />Exposure time: 2 minutes 34 seconds<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/HA721034" style="font-weight: bold;text-decoration: underline;">HA721034</a>) at 1/500 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 1:200,000 dilution was used for 1 hour at room temperature.

    ☑ Cell treatment (CT)

    Western blot analysis of Histone H3.3 (phospho S31) on Hela cell lysates.

    Lane 1: Hela cells, whole cell lysate, 10ug/lane
    Lane 2/3: Hela cells treated with 100ng/ml Nocodazole for 18h, whole cell lysates, 10ug/lane
    Lane 4: Hela cells treated with 100ng/ml Nocodazole for 18h, then treated with 2.8ug/ul lambda-PP for 30 minutes, whole cell lysates, 10ug/lane

    All lanes :
    Anti-Histone H3.3 (phospho S31) antibody (HA721034) at 1:500 dilution. Anti-GAPDH antibody (ET1601-4) at 1:10,000 dilution. Goat Anti-Rabbit IgG H&L (HRP) (HA1001) at 1/200,000 dilution.

    Predicted band size: 15 kDa
    Observed band size: 15 kDa

    Blocking and diluting buffer: 5% BSA.
    Exposure time: 2 minutes 34 seconds

    Proteins were transferred to a PVDF membrane and blocked with 5% NFDM/TBST for 1 hour at room temperature. The primary antibody (HA721034) at 1/500 dilution was used in 5% NFDM/TBST at room temperature for 2 hours. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1:200,000 dilution was used for 1 hour at room temperature.

  • <span style="font-weight: bold;">☑ Cell treatment (CT)</span><br /><br />Western blot analysis of Phospho-Histone H3.3 (S31) on different lysates with Rabbit anti-Phospho-Histone H3.3 (S31) antibody (<a href="/products/HA721034" style="font-weight: bold;text-decoration: underline;">HA721034</a>) at 1/1,000 dilution.<br /><br />Lane 1: PC-12 cell lysate (20 µg/Lane)<br />Lane 2: PC-12 treated with 100ng/mL Nocodazole for 18 hours cell lysate (20 µg/Lane)<br /><br />Predicted band size: 15 kDa<br />Observed band size: 15 kDa<br /><br />Exposure time: 1 minute;<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/HA721034" style="font-weight: bold;text-decoration: underline;">HA721034</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.

    ☑ Cell treatment (CT)

    Western blot analysis of Phospho-Histone H3.3 (S31) on different lysates with Rabbit anti-Phospho-Histone H3.3 (S31) antibody (HA721034) at 1/1,000 dilution.

    Lane 1: PC-12 cell lysate (20 µg/Lane)
    Lane 2: PC-12 treated with 100ng/mL Nocodazole for 18 hours cell lysate (20 µg/Lane)

    Predicted band size: 15 kDa
    Observed band size: 15 kDa

    Exposure time: 1 minute;
    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 (HA721034) 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.

  • <span style="font-weight: bold;">☑ Cell treatment (CT)</span><br /><br />Immunocytochemistry analysis of HeLa cells treated with or without 100ng/mL Nocodazole for 18 hours labeling Phospho-Histone H3.3 (S31) with Rabbit anti-Phospho-Histone H3.3 (S31) antibody (<a href="/products/HA721034" style="font-weight: bold;text-decoration: underline;">HA721034</a>) at 1/25,000 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-Phospho-Histone H3.3 (S31) antibody (<a href="/products/HA721034" style="font-weight: bold;text-decoration: underline;">HA721034</a>) at 1/25,000 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.

    ☑ Cell treatment (CT)

    Immunocytochemistry analysis of HeLa cells treated with or without 100ng/mL Nocodazole for 18 hours labeling Phospho-Histone H3.3 (S31) with Rabbit anti-Phospho-Histone H3.3 (S31) antibody (HA721034) at 1/25,000 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-Phospho-Histone H3.3 (S31) antibody (HA721034) at 1/25,000 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.

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

Citation