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eIF3A Recombinant Rabbit Monoclonal Antibody [JE58-60]

Usd: 385

Specification

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Catalog# HA721375

eIF3A Recombinant Rabbit Monoclonal Antibody [JE58-60]

Application

  • WB

  • IHC-P

  • IF-Cell

Reactivity

  • Human

  • Mouse

  • Rat

Conjugation

  • unconjugated

Overview

Product Name

eIF3A Recombinant Rabbit Monoclonal Antibody [JE58-60]

Antibody Type

Recombinant Rabbit monoclonal Antibody

Immunogen

Synthetic peptide within Human eIF3A aa 275-324 / 1,382.

Species Reactivity

Human, Mouse, Rat

Validated Applications

WB, IHC-P, IF-Cell

Target Molecular Weight

Predicted band size: 167 kDa

Positive Control

293T cell lysate, A431 cell lysate, Jurkat cell lysate, Raji cell lysate, RAW264.7 cell lysate, NIH/3T3 cell lysate, L6 cell lysate, PC-12 cell lysate, mouse brain tissue, rat brain tissue, Hela.

Conjugation

unconjugated

Clone Number

JE58-60

RRID

AB_3072492

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

  • 1:1,000

  • IHC-P

  • 1:1,000

  • IF-Cell

  • 1:100

Target

Function

Component of the eukaryotic translation initiation factor 3 (eIF-3) complex, which is required for several steps in the initiation of protein synthesis. The eIF-3 complex associates with the 40S ribosome and facilitates the recruitment of eIF-1, eIF-1A, eIF-2:GTP:methionyl-tRNAi and eIF-5 to form the 43S pre-initiation complex (43S PIC). The eIF-3 complex stimulates mRNA recruitment to the 43S PIC and scanning of the mRNA for AUG recognition. The eIF-3 complex is also required for disassembly and recycling of post-termination ribosomal complexes and subsequently prevents premature joining of the 40S and 60S ribosomal subunits prior to initiation. Essential for the initiation of translation on type-1 viral ribosomal entry sites (IRESs), like for HCV, PV, EV71 or BEV translation. In case of FCV infection, plays a role in the ribosomal termination-reinitiation event leading to the translation of VP2.

Background References

1. Lin L et al. Molecular interaction between human tumor marker protein p150, the largest subunit of eIF3, and intermediate filament protein K7. J. Cell. Biochem. 80:483-490 (2001)

2. Masutani M et al. Reconstitution reveals the functional core of mammalian eIF3. EMBO J. 26:3373-3383 (2007)

Subcellular Location

Cytoplasm.

UNIPROT

SwissProt: Q14152 Human

SwissProt: P23116 Mouse

SwissProt: Q1JU68 Rat

Synonyms

centrosomin homolog antibody

cytoplasmic protein p167 antibody

eIF 3 theta antibody

eIF-3-theta antibody

EIF3 antibody

eIF3 p167 antibody

eIF3 p170 antibody

eIF3 p180 antibody

EIF3 p180 subunit antibody

eIF3 p185 antibody

Expand

Images

  • Western blot analysis of eIF3A on different lysates with Rabbit anti-eIF3A antibody (<a href="/products/HA721375" style="font-weight: bold;text-decoration: underline;">HA721375</a>) at 1/1,000 dilution.<br /><br />Lane 1: 293T cell lysate<br />Lane 2: A431 cell lysate<br />Lane 3: Jurkat cell lysate<br />Lane 4: Raji cell lysate<br />Lane 5: RAW264.7 cell lysate<br />Lane 6: NIH/3T3 cell lysate<br />Lane 7: L6 cell lysate<br />Lane 8: PC-12 cell lysate<br /><br />Lysates/proteins at 15 µg/Lane.<br /><br />Predicted band size: 167 kDa<br />Observed band size: 167 kDa<br /><br />Exposure time: 7seconds;<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/HA721375" style="font-weight: bold;text-decoration: underline;">HA721375</a>) at 1/1,000 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 1:100,000 dilution was used for 1 hour at room temperature.

    Western blot analysis of eIF3A on different lysates with Rabbit anti-eIF3A antibody (HA721375) at 1/1,000 dilution.

    Lane 1: 293T cell lysate
    Lane 2: A431 cell lysate
    Lane 3: Jurkat cell lysate
    Lane 4: Raji cell lysate
    Lane 5: RAW264.7 cell lysate
    Lane 6: NIH/3T3 cell lysate
    Lane 7: L6 cell lysate
    Lane 8: PC-12 cell lysate

    Lysates/proteins at 15 µg/Lane.

    Predicted band size: 167 kDa
    Observed band size: 167 kDa

    Exposure time: 7seconds;

    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 (HA721375) at 1/1,000 dilution was used in 5% NFDM/TBST at room temperature for 2 hours. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1:100,000 dilution was used for 1 hour at room temperature.

  • Immunohistochemical analysis of paraffin-embedded mouse brain tissue with Rabbit anti-eIF3A antibody (<a href="/products/HA721375" style="font-weight: bold;text-decoration: underline;">HA721375</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/HA721375" style="font-weight: bold;text-decoration: underline;">HA721375</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 brain tissue with Rabbit anti-eIF3A antibody (HA721375) 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 (HA721375) 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 rat brain tissue with Rabbit anti-eIF3A antibody (<a href="/products/HA721375" style="font-weight: bold;text-decoration: underline;">HA721375</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/HA721375" style="font-weight: bold;text-decoration: underline;">HA721375</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 rat brain tissue with Rabbit anti-eIF3A antibody (HA721375) 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 (HA721375) 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.

  • Immunocytochemistry analysis of Hela cells labeling eIF3A with Rabbit anti-eIF3A antibody (<a href="/products/HA721375" style="font-weight: bold;text-decoration: underline;">HA721375</a>) at 1/100 dilution.<br /><br />Cells were fixed in 4% paraformaldehyde for 10 minutes at 37 ℃, permeabilized with 0.05% Triton X-100 in PBS for 20 minutes, and then blocked with 2% negative goat serum for 30 minutes at room temperature. Cells were then incubated with Rabbit anti-eIF3A antibody (<a href="/products/HA721375" style="font-weight: bold;text-decoration: underline;">HA721375</a>) at 1/100 dilution in 2% negative goat serum 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. 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 1/200 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 eIF3A with Rabbit anti-eIF3A antibody (HA721375) at 1/100 dilution.

    Cells were fixed in 4% paraformaldehyde for 10 minutes at 37 ℃, permeabilized with 0.05% Triton X-100 in PBS for 20 minutes, and then blocked with 2% negative goat serum for 30 minutes at room temperature. Cells were then incubated with Rabbit anti-eIF3A antibody (HA721375) at 1/100 dilution in 2% negative goat serum overnight at 4 ℃. Goat Anti-Rabbit IgG H&L (iFluor™ 488, HA1121) was used as the secondary antibody at 1/1,000 dilution. Nuclear DNA was labelled in blue with DAPI.

    Beta tubulin (M1305-2, red) was stained at 1/200 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 A431 cells labeling eIF3A with Rabbit anti-eIF3A antibody (<a href="/products/HA721375" style="font-weight: bold;text-decoration: underline;">HA721375</a>) at 1/100 dilution.<br /><br />Cells were fixed in 4% paraformaldehyde for 10 minutes at 37 ℃, permeabilized with 0.05% Triton X-100 in PBS for 20 minutes, and then blocked with 2% negative goat serum for 30 minutes at room temperature. Cells were then incubated with Rabbit anti-eIF3A antibody (<a href="/products/HA721375" style="font-weight: bold;text-decoration: underline;">HA721375</a>) at 1/100 dilution in 2% negative goat serum 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. 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 1/200 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 A431 cells labeling eIF3A with Rabbit anti-eIF3A antibody (HA721375) at 1/100 dilution.

    Cells were fixed in 4% paraformaldehyde for 10 minutes at 37 ℃, permeabilized with 0.05% Triton X-100 in PBS for 20 minutes, and then blocked with 2% negative goat serum for 30 minutes at room temperature. Cells were then incubated with Rabbit anti-eIF3A antibody (HA721375) at 1/100 dilution in 2% negative goat serum overnight at 4 ℃. Goat Anti-Rabbit IgG H&L (iFluor™ 488, HA1121) was used as the secondary antibody at 1/1,000 dilution. Nuclear DNA was labelled in blue with DAPI.

    Beta tubulin (M1305-2, red) was stained at 1/200 dilution overnight at +4℃. Goat Anti-Mouse IgG H&L (iFluor™ 594, HA1126) was used as the secondary antibody at 1/1,000 dilution.

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