Skip to content

Histone H3 (mutated K27 M) Recombinant Rabbit Monoclonal Antibody [JE46-51]

Usd: 385

Specification

Bulk Order Quote | Customization Request

Catalog# HA722200

Histone H3 (mutated K27 M) Recombinant Rabbit Monoclonal Antibody [JE46-51]

Application

  • WB

  • IF-Cell

  • IHC-P

  • ChIP

Reactivity

  • Human

Predicted reactivity

  • Mouse

Predicted species support after-sales service
Conjugation

  • unconjugated

Overview

Product Name

Histone H3 (mutated K27 M) Recombinant Rabbit Monoclonal Antibody [JE46-51]

Antibody Type

Recombinant Rabbit monoclonal Antibody

Immunogen

Synthetic peptide within Human Histone H3.3 aa 1-50 (mutated K27 M).

Species Reactivity

Human (Predicted: Mouse)

Predicted species support after-sales service

Validated Applications

WB, IF-Cell, IHC-P, ChIP

Molecular Weight

Predicted band size: 15 kDa

Positive Control

293T-OE-Histone H3 (mutated K27 M) cell lysate, HeLa cells transfected with Histone H3 (mutated K27 M).

Conjugation

unconjugated

Clone Number

JE46-51

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

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

  • 1:1,000-1:5,000

  • IF-Cell

  • 1:1,000

  • IHC-P

  • 1:1,000

  • ChIP

  • Use 0.5~2 μg for 25 μg of chromatin.

Target

Function

Histone H3 is one of the five main histones involved in the structure of chromatin in eukaryotic cells. Featuring a main globular domain and a long N-terminal tail, H3 is involved with the structure of the nucleosomes of the 'beads on a string' structure. Histone proteins are highly post-translationally modified however Histone H3 is the most extensively modified of the five histones. The term "Histone H3" alone is purposely ambiguous in that it does not distinguish between sequence variants or modification state. Histone H3 is an important protein in the emerging field of epigenetics, where its sequence variants and variable modification states are thought to play a role in the dynamic and long term regulation of genes.

Background References

1. Gang Wu et al. Somatic histone H3 alterations in pediatric diffuse intrinsic pontine gliomas and non-brainstem glioblastomas. Nat Genet. 2012 Jan

Subcellular Location

Nucleus, Chromosome.

UNIPROT

SwissProt: P68431 Human

SwissProt: P84243 Human

SwissProt: Q16695 Human

SwissProt: Q6NXT2 Human

SwissProt: Q71DI3 Human

Synonyms

H3 histone family member E pseudogene antibody

H3 histone family, member A antibody

H3/A antibody

H31_HUMAN antibody

H3F3 antibody

H3FA antibody

Hist1h3a antibody

HIST1H3B antibody

HIST1H3C antibody

HIST1H3D antibody

Expand

Images

  • <span style="font-weight: bold;">☑ Cell treatment (CT)</span><br /><br />Western blot analysis of Histone H3 (mutated K27 M) on different lysates with Rabbit anti-Histone H3 (mutated K27 M) antibody (<a href="/products/HA722200" style="font-weight: bold;text-decoration: underline;">HA722200</a>) at 1/1,000 dilution.<br /><br />Lane 1: 293T-OE-Histone H3 (mutated K27 M) cell lysate<br />Lane 2: 293T-OE-Histone H3 cell lysate (negative)<br /><br />Lysates/proteins at 10 µg/Lane.<br /><br />Predicted band size: 15 kDa<br />Observed band size: 18 kDa<br /><br />Exposure time: 2 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/HA722200" style="font-weight: bold;text-decoration: underline;">HA722200</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 Histone H3 (mutated K27 M) on different lysates with Rabbit anti-Histone H3 (mutated K27 M) antibody (HA722200) at 1/1,000 dilution.

    Lane 1: 293T-OE-Histone H3 (mutated K27 M) cell lysate
    Lane 2: 293T-OE-Histone H3 cell lysate (negative)

    Lysates/proteins at 10 µg/Lane.

    Predicted band size: 15 kDa
    Observed band size: 18 kDa

    Exposure time: 2 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 (HA722200) 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;">☑ Relative expression (RE)</span><br /><br />Western blot analysis of Histone H3 (mutated K27 M) on different lysates with Rabbit anti-Histone H3 (mutated K27 M) antibody (<a href="/products/HA722200" style="font-weight: bold;text-decoration: underline;">HA722200</a>) at 1/2,000 dilution.<br /><br />Lane 1: HeLa cell lysate (negative)<br />Lane 2: Raji cell lysate (negative)<br />Lane 3: 293T cell lysate (negative)<br />Lane 4: HT-29 cell lysate (negative)<br />Lane 5: Jurkat cell lysate (negative)<br /><br />Lysates/proteins at 30 µg/Lane.<br /><br />Exposure time: 3 minutes; 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/HA722200" style="font-weight: bold;text-decoration: underline;">HA722200</a>) at 1/2,000 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.

    ☑ Relative expression (RE)

    Western blot analysis of Histone H3 (mutated K27 M) on different lysates with Rabbit anti-Histone H3 (mutated K27 M) antibody (HA722200) at 1/2,000 dilution.

    Lane 1: HeLa cell lysate (negative)
    Lane 2: Raji cell lysate (negative)
    Lane 3: 293T cell lysate (negative)
    Lane 4: HT-29 cell lysate (negative)
    Lane 5: Jurkat cell lysate (negative)

    Lysates/proteins at 30 µg/Lane.

    Exposure time: 3 minutes; 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 (HA722200) at 1/2,000 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 transfected with Histone H3 / Histone H3 (mutated K27 M) labeling Histone H3 (mutated K27 M) with Rabbit anti-Histone H3 (mutated K27 M) antibody (<a href="/products/HA722200" style="font-weight: bold;text-decoration: underline;">HA722200</a>) at 1/1,000 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-Histone H3 (mutated K27 M) antibody (<a href="/products/HA722200" style="font-weight: bold;text-decoration: underline;">HA722200</a>) at 1/1,000 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 transfected with Histone H3 / Histone H3 (mutated K27 M) labeling Histone H3 (mutated K27 M) with Rabbit anti-Histone H3 (mutated K27 M) antibody (HA722200) at 1/1,000 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-Histone H3 (mutated K27 M) antibody (HA722200) at 1/1,000 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.

  • <span style="font-weight: bold;">☑ Cell treatment (CT)</span><br /><br />Immunohistochemical analysis of paraffin-embedded 293T cells transfected with Histone H3 / Histone H3 (mutated K27 M) with Rabbit anti-Histone H3 (mutated K27 M) antibody (<a href="/products/HA722200" style="font-weight: bold;text-decoration: underline;">HA722200</a>) at 1/1,000 dilution.<br /><br />The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) (high pressure) for 2 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH<sub>2</sub>O and PBS, and then probed with the primary antibody (<a href="/products/HA722200" style="font-weight: bold;text-decoration: underline;">HA722200</a>) at 1/1,000 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.

    ☑ Cell treatment (CT)

    Immunohistochemical analysis of paraffin-embedded 293T cells transfected with Histone H3 / Histone H3 (mutated K27 M) with Rabbit anti-Histone H3 (mutated K27 M) antibody (HA722200) at 1/1,000 dilution.

    The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) (high pressure) for 2 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (HA722200) at 1/1,000 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.

  • <span style="font-weight: bold;">☑ Cell treatment (CT)</span><br /><br />Chromatin immunoprecipitations were performed with cross-linked chromatin from 293T cells transfected with Histone H3 (mutated K27 M) cells with Histone H3 (mutated K27 M) (<a href="/products/HA722200" style="font-weight: bold;text-decoration: underline;">HA722200</a>) or Normal Rabbit IgG according to the ChIP protocol. The enriched DNA was quantified by real-time PCR using indicated primers. The amount of immunoprecipitated DNA in each sample is represented as signal relative to the total amount of input chromatin, which is equivalent to one.

    ☑ Cell treatment (CT)

    Chromatin immunoprecipitations were performed with cross-linked chromatin from 293T cells transfected with Histone H3 (mutated K27 M) cells with Histone H3 (mutated K27 M) (HA722200) or Normal Rabbit IgG according to the ChIP protocol. The enriched DNA was quantified by real-time PCR using indicated primers. The amount of immunoprecipitated DNA in each sample is represented as signal relative to the total amount of input chromatin, which is equivalent to one.

  • <span style="font-weight: bold;">☑ Cell treatment (CT)</span><br /><br />Chromatin immunoprecipitations were performed with cross-linked chromatin from 293T cells transfected with Histone H3 cells (negative) with Histone H3 (mutated K27 M) (<a href="/products/HA722200" style="font-weight: bold;text-decoration: underline;">HA722200</a>) or Normal Rabbit IgG according to the ChIP protocol. The enriched DNA was quantified by real-time PCR using indicated primers. The amount of immunoprecipitated DNA in each sample is represented as signal relative to the total amount of input chromatin, which is equivalent to one.

    ☑ Cell treatment (CT)

    Chromatin immunoprecipitations were performed with cross-linked chromatin from 293T cells transfected with Histone H3 cells (negative) with Histone H3 (mutated K27 M) (HA722200) or Normal Rabbit IgG according to the ChIP protocol. The enriched DNA was quantified by real-time PCR using indicated primers. The amount of immunoprecipitated DNA in each sample is represented as signal relative to the total amount of input chromatin, which is equivalent to one.

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