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USF1 Recombinant Rabbit Monoclonal Antibody [JE63-68]

Usd: 385

Specification

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Catalog# HA722469

USF1 Recombinant Rabbit Monoclonal Antibody [JE63-68]

Application

  • WB

  • IHC-P

  • IF-Cell

  • FC

Reactivity

  • Human

  • Mouse

  • Rat

Conjugation

  • unconjugated

Overview

Product Name

USF1 Recombinant Rabbit Monoclonal Antibody [JE63-68]

Antibody Type

Recombinant Rabbit monoclonal Antibody

Immunogen

Synthetic peptide within Human USF1 aa 150-250.

Species Reactivity

Human, Mouse, Rat

Validated Applications

WB, IHC-P, IF-Cell, FC

Molecular Weight

Predicted band size: 34 kDa

Positive Control

HEK-293 cell lysate, K-562 cell lysate, NIH/3T3 cell lysate, Mouse kidney tissue lysate, Rat kidney tissue lysate, HeLa, mouse bladder tissue, rat bladder tissue.

Conjugation

unconjugated

Clone Number

JE63-68

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

  • IHC-P

  • 1:1,000

  • IF-Cell

  • 1:50

  • FC

  • 1:1,000

Target

Function

This gene encodes a member of the basic helix-loop-helix leucine zipper family, and can function as a cellular transcription factor. The encoded protein can activate transcription through pyrimidine-rich initiator (Inr) elements and E-box motifs. This gene has been linked to familial combined hyperlipidemia (FCHL). Alternative splicing of this gene results in multiple transcript variants. A related pseudogene has been defined on chromosome 21. A disorder characterized by a variable pattern of elevated levels of serum total cholesterol, triglycerides or both. It is observed in a percentage of individuals with premature coronary heart disease.

Background References

1. Wang D., Jin X., Lei M., Jiang Y., Liu Y., Yu F., Guo Y., Han B., Yang Y., Chen X. USF1-ATRAP-PBX3 Axis Promote Breast Cancer Glycolysis and Malignant Phenotype by Activating AKT/mTOR Signaling. Int J Biol Sci 18:2452-2471 (2022)

2. Alper M., Aydemir T., Kockar F. [USF1 Suppresses Expression of Fibrillar Type I, II, and III Collagen and pNP Adamts-3 in Osteosarcoma Cells]. Mol Biol (Mosk) 55:634-642 (2021)

Subcellular Location

Nucleus.

UNIPROT

SwissProt: P22415 Human

SwissProt: Q61069 Mouse

Entrez Gene: 83586 Rat

Synonyms

bHLHb11 antibody

Class B basic helix-loop-helix protein 11 antibody

FCHL 1 antibody

FCHL antibody

FCHL1 antibody

HGNC:12593 antibody

HYPLIP 1 antibody

HYPLIP1 antibody

Major late transcription factor 1 antibody

Major late transcription factor antibody

Expand

Images

  • Western blot analysis of USF1 on different lysates with Rabbit anti-USF1 antibody (<a href="/products/HA722469" style="font-weight: bold;text-decoration: underline;">HA722469</a>) at 1/1,000 dilution.<br /><br />Lane 1: HEK-293 cell lysate (20 µg/Lane)<br />Lane 2: K-562 cell lysate (20 µg/Lane)<br />Lane 3: NIH/3T3 cell lysate (20 µg/Lane)<br />Lane 4: Mouse kidney tissue lysate (40 µg/Lane)<br />Lane 5: Rat kidney tissue lysate (40 µg/Lane)<br /><br />Predicted band size: 34 kDa<br />Observed band size: 34 kDa<br /><br />Exposure time: 6 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/HA722469" style="font-weight: bold;text-decoration: underline;">HA722469</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 USF1 on different lysates with Rabbit anti-USF1 antibody (HA722469) at 1/1,000 dilution.

    Lane 1: HEK-293 cell lysate (20 µg/Lane)
    Lane 2: K-562 cell lysate (20 µg/Lane)
    Lane 3: NIH/3T3 cell lysate (20 µg/Lane)
    Lane 4: Mouse kidney tissue lysate (40 µg/Lane)
    Lane 5: Rat kidney tissue lysate (40 µg/Lane)

    Predicted band size: 34 kDa
    Observed band size: 34 kDa

    Exposure time: 6 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 (HA722469) 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 USF1 with Rabbit anti-USF1 antibody (<a href="/products/HA722469" style="font-weight: bold;text-decoration: underline;">HA722469</a>) at 1/50 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-USF1 antibody (<a href="/products/HA722469" style="font-weight: bold;text-decoration: underline;">HA722469</a>) at 1/50 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 USF1 with Rabbit anti-USF1 antibody (HA722469) at 1/50 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-USF1 antibody (HA722469) at 1/50 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 mouse bladder tissue with Rabbit anti-USF1 antibody (<a href="/products/HA722469" style="font-weight: bold;text-decoration: underline;">HA722469</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/HA722469" style="font-weight: bold;text-decoration: underline;">HA722469</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 bladder tissue with Rabbit anti-USF1 antibody (HA722469) 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 (HA722469) 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 bladder tissue with Rabbit anti-USF1 antibody (<a href="/products/HA722469" style="font-weight: bold;text-decoration: underline;">HA722469</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/HA722469" style="font-weight: bold;text-decoration: underline;">HA722469</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 bladder tissue with Rabbit anti-USF1 antibody (HA722469) 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 (HA722469) 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 USF1.<br /><br />Cells were fixed and permeabilized. Then stained with the primary antibody (<a href="/products/HA722469" style="font-weight: bold;text-decoration: underline;">HA722469</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 USF1.

    Cells were fixed and permeabilized. Then stained with the primary antibody (HA722469, 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).

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