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beta Actin Recombinant Rabbit Monoclonal Antibody [PSH03-63]

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Catalog# HA722023

beta Actin Recombinant Rabbit Monoclonal Antibody [PSH03-63]

Application

  • WB

  • IF-Cell

  • FC

  • IP

Reactivity

  • Human

  • Mouse

  • Rat

BSA and Azide free
Predicted reactivity

  • Rabbit

  • Goat

  • Dog

  • Monkey

  • Pig

  • Chicken

  • Cow

  • Hamster

  • Green monkey

予測される種属に対するアフターサービスのサポート
Conjugation

  • unconjugated

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

概要

製品名

beta Actin Recombinant Rabbit Monoclonal Antibody [PSH03-63]

抗体の種類

Recombinant Rabbit monoclonal Antibody

免疫原

Synthetic peptide within N-terminal residues of β-Actin.

種属反応性

Human, Mouse, Rat (Predicted: Rabbit, Goat, Dog, Monkey, Pig, Chicken, Cow, Hamster, Green monkey)

予測される種属に対するアフターサービスのサポート

検証された応用例

WB, IF-Cell, FC, IP

分子量

Predicted band size: 42 kDa

陽性対照

HeLa cell lysate, HepG2 cell lysate, MCF7 cell lysate, A431 cell lysate, Jurkat cell lysate, HEK-293 cell lysate, RAW264.7 cell lysate, C2C12 cell lysate, PC-12 cell lysate, mouse testis tissue lysate, mouse spleen tissue lysate, rat testis tissue lysate, rat kidney tissue lysate, HeLa, NIH/3T3, C6.

結合

unconjugated

クローン番号

PSH03-63

RRID

AB_3096833

製品の特徴

形態

Liquid

濃度

保存方法

Shipped at 4℃. Store at +4℃ short term (1-2 weeks). It is recommended to aliquot into single-use upon delivery. Store at -20℃ long term.

保存用バッファー

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

アイソタイプ

IgG

精製方法

Protein A affinity purified.

応用希釈度

  • WB

  • 1:20,000-1:100,000

  • IF-Cell

  • 1:100-1:250

  • FC

  • 1:1,000

  • IP

  • 1-2μg/sample

ターゲット

機能

Actins are highly conserved proteins involved in cell motility, structure and integrity. Actin has been found to be expressed in at least six isomeric forms. It is expressed in heart and skeletal striated muscle tissue, and in certain smooth muscle tissues, regulating contractile potentials for these cells. It is also expressed in the cytoplasm of non-muscle cells, functioning to control cell structure and motility. Beta actin is usually used as a loading control, for among others, the integrity of cells, protein degradation, in Western Blotting.

背景文献

1. Ponte P., Ng S.Y., Engel J., Gunning P., Kedes L."Evolutionary conservation in the untranslated regions of actin mRNAs: DNA sequence of a human beta-actin cDNA." Nucleic Acids Res. 12:1687-1696(1984) Ohmori H., Toyama S., Toyama S."Direct proof that the primary site of action of cytochalasin on cell motility processes is actin."J. Cell Biol. 116:933-941(1992)

亜細胞局在

Cytoskeleton.

UNIPROT #

SwissProt: P60709 Human

SwissProt: P60710 Mouse

SwissProt: P60711 Rat

別名

A26C1A antibody

A26C1B antibody

ACTB antibody

ACTB_HUMAN antibody

Actin beta antibody

Actin cytoplasmic 1 antibody

Actin, cytoplasmic 1, N-terminally processed antibody

Actx antibody

b actin antibody

Beta cytoskeletal actin antibody

展開

画像

  • Western blot analysis of beta Actin on different lysates with Rabbit anti-beta Actin antibody (<a href="/products/HA722023" style="font-weight: bold;text-decoration: underline;">HA722023</a>) at 1/20,000 dilution.<br /><br />Lane 1: HeLa cell lysate (10 µg/Lane)<br />Lane 2: HepG2 cell lysate (10 µg/Lane)<br />Lane 3: MCF7 cell lysate (10 µg/Lane)<br />Lane 4: A431 cell lysate (10 µg/Lane)<br />Lane 5: Jurkat cell lysate (10 µg/Lane)<br />Lane 6: HEK-293 cell lysate (10 µg/Lane)<br />Lane 7: RAW264.7 cell lysate (10 µg/Lane)<br />Lane 8: C2C12 cell lysate (10 µg/Lane)<br />Lane 9: PC-12 cell lysate (10 µg/Lane)<br />Lane 10: Mouse testis tissue lysate (10 µg/Lane)<br />Lane 11: Mouse spleen tissue lysate (10 µg/Lane)<br />Lane 12: Rat testis tissue lysate (10 µg/Lane)<br />Lane 13: Rat kidney tissue lysate (10 µg/Lane)<br /><br />Predicted band size: 42 kDa<br />Observed band size: 42 kDa<br /><br />Exposure time: 3 seconds;<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/HA722023" style="font-weight: bold;text-decoration: underline;">HA722023</a>) at 1/20,000 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 beta Actin on different lysates with Rabbit anti-beta Actin antibody (HA722023) at 1/20,000 dilution.

    Lane 1: HeLa cell lysate (10 µg/Lane)
    Lane 2: HepG2 cell lysate (10 µg/Lane)
    Lane 3: MCF7 cell lysate (10 µg/Lane)
    Lane 4: A431 cell lysate (10 µg/Lane)
    Lane 5: Jurkat cell lysate (10 µg/Lane)
    Lane 6: HEK-293 cell lysate (10 µg/Lane)
    Lane 7: RAW264.7 cell lysate (10 µg/Lane)
    Lane 8: C2C12 cell lysate (10 µg/Lane)
    Lane 9: PC-12 cell lysate (10 µg/Lane)
    Lane 10: Mouse testis tissue lysate (10 µg/Lane)
    Lane 11: Mouse spleen tissue lysate (10 µg/Lane)
    Lane 12: Rat testis tissue lysate (10 µg/Lane)
    Lane 13: Rat kidney tissue lysate (10 µg/Lane)

    Predicted band size: 42 kDa
    Observed band size: 42 kDa

    Exposure time: 3 seconds;
    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 (HA722023) at 1/20,000 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.

  • Western blot analysis of beta Actin on HepG2 cell lysates with Rabbit anti-beta Actin antibody (<a href="/products/HA722023" style="font-weight: bold;text-decoration: underline;">HA722023</a>) at different dilutions.<br /><br />Lysates/proteins at 20 µg/Lane.<br /><br />Predicted band size: 42 kDa<br />Observed band size: 42 kDa<br /><br />Exposure time: 30 seconds;<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/HA722023" style="font-weight: bold;text-decoration: underline;">HA722023</a>) at different dilutions 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 beta Actin on HepG2 cell lysates with Rabbit anti-beta Actin antibody (HA722023) at different dilutions.

    Lysates/proteins at 20 µg/Lane.

    Predicted band size: 42 kDa
    Observed band size: 42 kDa

    Exposure time: 30 seconds;
    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 (HA722023) at different dilutions 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 beta Actin with Rabbit anti-beta Actin antibody (<a href="/products/HA722023" style="font-weight: bold;text-decoration: underline;">HA722023</a>) at 1/250 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-beta Actin antibody (<a href="/products/HA722023" style="font-weight: bold;text-decoration: underline;">HA722023</a>) at 1/250 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 beta Actin with Rabbit anti-beta Actin antibody (HA722023) at 1/250 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-beta Actin antibody (HA722023) at 1/250 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 beta Actin with Rabbit anti-beta Actin antibody (<a href="/products/HA722023" style="font-weight: bold;text-decoration: underline;">HA722023</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-beta Actin antibody (<a href="/products/HA722023" style="font-weight: bold;text-decoration: underline;">HA722023</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 beta Actin with Rabbit anti-beta Actin antibody (HA722023) 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-beta Actin antibody (HA722023) 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 L6 cells labeling beta Actin with Rabbit anti-beta Actin antibody (<a href="/products/HA722023" style="font-weight: bold;text-decoration: underline;">HA722023</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-beta Actin antibody (<a href="/products/HA722023" style="font-weight: bold;text-decoration: underline;">HA722023</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 L6 cells labeling beta Actin with Rabbit anti-beta Actin antibody (HA722023) 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-beta Actin antibody (HA722023) 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 beta Actin.<br /><br />Cells were fixed and permeabilized. Then stained with the primary antibody (<a href="/products/HA722023" style="font-weight: bold;text-decoration: underline;">HA722023</a>, 1μg/mL) (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 beta Actin.

    Cells were fixed and permeabilized. Then stained with the primary antibody (HA722023, 1μg/mL) (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).

  • Flow cytometric analysis of NIH/3T3 cells labeling beta Actin.<br /><br />Cells were fixed and permeabilized. Then stained with the primary antibody (<a href="/products/HA722023" style="font-weight: bold;text-decoration: underline;">HA722023</a>, 1μg/mL) (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 NIH/3T3 cells labeling beta Actin.

    Cells were fixed and permeabilized. Then stained with the primary antibody (HA722023, 1μg/mL) (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).

  • Flow cytometric analysis of C6 cells labeling beta Actin.<br /><br />Cells were fixed and permeabilized. Then stained with the primary antibody (<a href="/products/HA722023" style="font-weight: bold;text-decoration: underline;">HA722023</a>, 1μg/mL) (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 C6 cells labeling beta Actin.

    Cells were fixed and permeabilized. Then stained with the primary antibody (HA722023, 1μg/mL) (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).

  • beta Actin was immunoprecipitated from 0.2 mg NIH/3T3 cell lysate with <a href="/products/HA722023" style="font-weight: bold;text-decoration: underline;">HA722023</a> at 2 µg/10 µl beads. Western blot was performed from the immunoprecipitate using <a href="/products/HA722023" style="font-weight: bold;text-decoration: underline;">HA722023</a> at 1/5,000 dilution. Anti-Rabbit IgG for IP Nano-secondary antibody (NBI01H) at 1/5,000 dilution was used for 1 hour at room temperature.<br /><br />Lane 1: NIH/3T3 cell lysate (input)<br />Lane 2: <a href="/products/HA722023" style="font-weight: bold;text-decoration: underline;">HA722023</a> IP in NIH/3T3 cell lysate<br />Lane 3: Rabbit IgG instead of <a href="/products/HA722023" style="font-weight: bold;text-decoration: underline;">HA722023</a> in NIH/3T3 cell lysate<br /><br />Blocking/Dilution buffer: 5% NFDM/TBST<br />Exposure time: 1 minute 2 seconds; ECL: K1801

    beta Actin was immunoprecipitated from 0.2 mg NIH/3T3 cell lysate with HA722023 at 2 µg/10 µl beads. Western blot was performed from the immunoprecipitate using HA722023 at 1/5,000 dilution. Anti-Rabbit IgG for IP Nano-secondary antibody (NBI01H) at 1/5,000 dilution was used for 1 hour at room temperature.

    Lane 1: NIH/3T3 cell lysate (input)
    Lane 2: HA722023 IP in NIH/3T3 cell lysate
    Lane 3: Rabbit IgG instead of HA722023 in NIH/3T3 cell lysate

    Blocking/Dilution buffer: 5% NFDM/TBST
    Exposure time: 1 minute 2 seconds; ECL: K1801

ご注意ください: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"

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