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PABP Recombinant Rabbit Monoclonal Antibody [PSH05-51]

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Specification

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Catalog# HA722289

PABP Recombinant Rabbit Monoclonal Antibody [PSH05-51]

Application

  • WB

  • IF-Cell

  • IHC-P

  • FC

  • IF-Tissue

Reactivity

  • Human

  • Mouse

  • Green monkey

BSA and Azide free
Conjugation

  • unconjugated

Overview

Product Name

PABP Recombinant Rabbit Monoclonal Antibody [PSH05-51]

Antibody Type

Recombinant Rabbit monoclonal Antibody

Immunogen

Recombinant protein within Human PABP protein aa 450-550.

Species Reactivity

Human, Mouse, Green monkey

Validated Applications

WB, IF-Cell, IHC-P, FC, IF-Tissue

Target Molecular Weight

Predicted band size: 71 kDa

Positive Control

TF-1 cell lysate, HeLa cell lysate, A549 cell lysate, MCF7 cell lysate, HepG2 cell lysate, HCT 116 cell lysate, HEK-293 cell lysate, NIH/3T3 cell lysate, COS-1 cell lysate, Vero cell lysate, mouse brain tissue lysate, HeLa, NIH/3T3, human colon tissue, human testis tissue, mouse testis tissue.

Conjugation

unconjugated

Clone Number

PSH05-51

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

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

  • IHC-P

  • 1:1,000

  • FC

  • 1:1,000

  • IF-Tissue

  • 1:200

Target

Function

The poly(A)-binding protein (PAB or PABP), which is found complexed to the 3' poly(A) tail of eukaryotic mRNA, is required for poly(A) lengthening and the termination of translation. In humans, the PABPs comprise a small nuclear isoform and a conserved gene family of other poly(A)-binding proteins. PABPC1 is usually diffused within the cytoplasm and concentrated at sites of high mRNA concentration such as stress granules, processing bodies, and locations of high translational activity. PABPC1 is also associated with nonsense-mediated mRNA decay (NMD). PABPC1 binds to the poly(A) tail and interact with eIF4G, which stabilizes the circularization of mRNAs. This structure is required for the prevention of mRNA degradation via NMD. In the nucleus PABP1 binds to the poly(A) tails of pre-mRNAs to facilitate stability, export, transport, and degradation. PABP1 binding is also required for nuclear-mediated degradation. PABPC1 contains four RNA-recognition motifs (RRMs). The first two, RRM1 and RRM2, bind both α-importin and the poly(A) tail of processed mRNA. This feature prevents mRNA from going back into the nucleus.

Background References

1. Wang J et al. PABP/purine-rich motif as an initiation module for cap-independent translation in pattern-triggered immunity. Cell. 2022 Aug

2. Kajjo S et al. PABP prevents the untimely decay of select mRNA populations in human cells. EMBO J. 2022 Mar

Subcellular Location

Cytoplasm, Stress granule, Nucleus, Cell projection, lamellipodium.

UNIPROT

SwissProt: P11940 Human

SwissProt: P29341 Mouse

Synonyms

PAB 1 antibody

PAB1 antibody

PABP 1 antibody

PABP-1 antibody

PABP1 antibody

PABP1_HUMAN antibody

PABPC 1 antibody

Pabpc1 antibody

PABPC2 antibody

PABPL1 antibody

Expand

Images

  • Western blot analysis of PABP on different lysates with Rabbit anti-PABP antibody (<a href="/products/HA722289" style="font-weight: bold;text-decoration: underline;">HA722289</a>) at 1/1,000 dilution.<br /><br />Lane 1: TF-1 cell lysate (20 µg/Lane)<br />Lane 2: HeLa cell lysate (20 µg/Lane)<br />Lane 3: A549 cell lysate (20 µg/Lane)<br />Lane 4: MCF7 cell lysate (20 µg/Lane)<br />Lane 5: HepG2 cell lysate (20 µg/Lane)<br />Lane 6: HCT 116 cell lysate (20 µg/Lane)<br />Lane 7: HEK-293 cell lysate (20 µg/Lane)<br />Lane 8: NIH/3T3 cell lysate (20 µg/Lane)<br />Lane 9: COS-1 cell lysate (20 µg/Lane)<br />Lane 10: Vero cell lysate (20 µg/Lane)<br />Lane 11: Mouse brain tissue lysate (40 µg/Lane)<br /><br />Predicted band size: 71 kDa<br />Observed band size: 71 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/HA722289" style="font-weight: bold;text-decoration: underline;">HA722289</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 PABP on different lysates with Rabbit anti-PABP antibody (HA722289) at 1/1,000 dilution.

    Lane 1: TF-1 cell lysate (20 µg/Lane)
    Lane 2: HeLa cell lysate (20 µg/Lane)
    Lane 3: A549 cell lysate (20 µg/Lane)
    Lane 4: MCF7 cell lysate (20 µg/Lane)
    Lane 5: HepG2 cell lysate (20 µg/Lane)
    Lane 6: HCT 116 cell lysate (20 µg/Lane)
    Lane 7: HEK-293 cell lysate (20 µg/Lane)
    Lane 8: NIH/3T3 cell lysate (20 µg/Lane)
    Lane 9: COS-1 cell lysate (20 µg/Lane)
    Lane 10: Vero cell lysate (20 µg/Lane)
    Lane 11: Mouse brain tissue lysate (40 µg/Lane)

    Predicted band size: 71 kDa
    Observed band size: 71 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 (HA722289) 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 PABP with Rabbit anti-PABP antibody (<a href="/products/HA722289" style="font-weight: bold;text-decoration: underline;">HA722289</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-PABP antibody (<a href="/products/HA722289" style="font-weight: bold;text-decoration: underline;">HA722289</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 PABP with Rabbit anti-PABP antibody (HA722289) 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-PABP antibody (HA722289) 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 PABP with Rabbit anti-PABP antibody (<a href="/products/HA722289" style="font-weight: bold;text-decoration: underline;">HA722289</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-PABP antibody (<a href="/products/HA722289" style="font-weight: bold;text-decoration: underline;">HA722289</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 PABP with Rabbit anti-PABP antibody (HA722289) 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-PABP antibody (HA722289) 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.

  • Immunohistochemical analysis of paraffin-embedded human colon tissue with Rabbit anti-PABP antibody (<a href="/products/HA722289" style="font-weight: bold;text-decoration: underline;">HA722289</a>) at 1/1,000 dilution.<br /><br />The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 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/HA722289" style="font-weight: bold;text-decoration: underline;">HA722289</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.

    Immunohistochemical analysis of paraffin-embedded human colon tissue with Rabbit anti-PABP antibody (HA722289) at 1/1,000 dilution.

    The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 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 (HA722289) 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.

  • Immunohistochemical analysis of paraffin-embedded human testis tissue with Rabbit anti-PABP antibody (<a href="/products/HA722289" style="font-weight: bold;text-decoration: underline;">HA722289</a>) at 1/1,000 dilution.<br /><br />The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 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/HA722289" style="font-weight: bold;text-decoration: underline;">HA722289</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.

    Immunohistochemical analysis of paraffin-embedded human testis tissue with Rabbit anti-PABP antibody (HA722289) at 1/1,000 dilution.

    The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 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 (HA722289) 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.

  • Immunohistochemical analysis of paraffin-embedded mouse testis tissue with Rabbit anti-PABP antibody (<a href="/products/HA722289" style="font-weight: bold;text-decoration: underline;">HA722289</a>) at 1/1,000 dilution.<br /><br />The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 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/HA722289" style="font-weight: bold;text-decoration: underline;">HA722289</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.

    Immunohistochemical analysis of paraffin-embedded mouse testis tissue with Rabbit anti-PABP antibody (HA722289) at 1/1,000 dilution.

    The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 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 (HA722289) 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.

  • Flow cytometric analysis of HeLa cells labeling PABP.<br /><br />Cells were fixed and permeabilized. Then stained with the primary antibody (<a href="/products/HA722289" style="font-weight: bold;text-decoration: underline;">HA722289</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 PABP.

    Cells were fixed and permeabilized. Then stained with the primary antibody (HA722289, 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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