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LAP3 Recombinant Rabbit Monoclonal Antibody [PSH01-26]

Usd: 385

Specification

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Catalog# HA721650

LAP3 Recombinant Rabbit Monoclonal Antibody [PSH01-26]

Application

  • WB

  • IHC-P

Reactivity

  • Human

  • Mouse

  • Rat

BSA and Azide free
Conjugation

  • unconjugated

Overview

Product Name

LAP3 Recombinant Rabbit Monoclonal Antibody [PSH01-26]

Antibody Type

Recombinant Rabbit monoclonal Antibody

Immunogen

Recombinant protein within human LAP3 aa 1-519 / 519.

Species Reactivity

Human, Mouse, Rat

Validated Applications

WB, IHC-P

Molecular Weight

Predicted band size: 56 kDa

Positive Control

HeLa cell lysate, HepG2 cell lysate, U-87 MG cell lysate, Daudi cell lysate, C6 cell lysate, mouse liver tissue lysate, rat liver tissue lysate, human liver tissue, HEK-293 cell lysate, K-562 cell lysate, A549 cell lysate, LO2 cell lysate, human testis tissue, mouse liver tissue.

Conjugation

unconjugated

Clone Number

PSH01-26

RRID

AB_3072763

Reactivity Data

WB IHC-P
Human
Tested Tested
Tested Tested
Mouse
Tested Tested
Tested Tested
Rat
Tested Tested

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

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

Isotype

IgG

Purification Method

Protein A affinity purified.

Application Dilution

  • WB

  • 1:1,000-1:5,000

  • IHC-P

  • 1:1,000

Target

Function

Cytosolic metallopeptidase that catalyzes the removal of unsubstituted N-terminal hydrophobic amino acids from various peptides. The presence of Zn(2+) ions is essential for the peptidase activity, and the association with other cofactors can modulate the substrate spectificity of the enzyme. For instance, in the presence of Mn(2+), it displays a specific Cys-Gly hydrolyzing activity of Cys-Gly-S-conjugates. Involved in the metabolism of glutathione and in the degradation of glutathione S-conjugates, which may play a role in the control of the cell redox status.

Background References

1. Li L et al. LAP3 contributes to IFN-γ-induced arginine depletion and malignant transformation of bovine mammary epithelial cells. BMC Cancer. 2022 Aug

2. Feng L et al. Cholesterol-induced leucine aminopeptidase 3 (LAP3) upregulation inhibits cell autophagy in pathogenesis of NAFLD. Aging (Albany NY). 2022 Apr

Subcellular Location

Cytoplasm

UNIPROT

SwissProt: P28838 Human

SwissProt: Q9CPY7 Mouse

SwissProt: Q68FS4 Rat

Synonyms

AMPL_HUMAN antibody

Cytosol aminopeptidase antibody

epididymis secretory protein Li 106 antibody

HEL-S-106 antibody

LAP 3 antibody

LAP antibody

LAP-3 antibody

Lap3 antibody

LAPEP antibody

Leucine aminopeptidase 3 antibody

Expand

Images

  • Western blot analysis of LAP3 on different lysates with Rabbit anti-LAP3 antibody (<a href="/products/HA721650" style="font-weight: bold;text-decoration: underline;">HA721650</a>) at 1/5,000 dilution and competitor's antibody at 1/1,000 dilution.<br /><br />Lane 1: HeLa cell lysate (15 µg/Lane)<br />Lane 2: HepG2 cell lysate (15 µg/Lane)<br />Lane 3: U-87 MG cell lysate (15 µg/Lane)<br />Lane 4: Daudi cell lysate (15 µg/Lane)<br />Lane 5: C6 cell lysate (15 µg/Lane)<br />Lane 6: Mouse liver tissue lysate (30 µg/Lane)<br />Lane 7: Rat liver tissue lysate (30 µg/Lane)<br /><br />Predicted band size: 56 kDa<br />Observed band size: 50 kDa<br /><br />Exposure time: Lane 1-7 (left): 1 minute; Lane 1-7 (right): 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/HA721650" style="font-weight: bold;text-decoration: underline;">HA721650</a>) at 1/5,000 dilution and competitor's antibody at 1/1,000 dilution were 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 LAP3 on different lysates with Rabbit anti-LAP3 antibody (HA721650) at 1/5,000 dilution and competitor's antibody at 1/1,000 dilution.

    Lane 1: HeLa cell lysate (15 µg/Lane)
    Lane 2: HepG2 cell lysate (15 µg/Lane)
    Lane 3: U-87 MG cell lysate (15 µg/Lane)
    Lane 4: Daudi cell lysate (15 µg/Lane)
    Lane 5: C6 cell lysate (15 µg/Lane)
    Lane 6: Mouse liver tissue lysate (30 µg/Lane)
    Lane 7: Rat liver tissue lysate (30 µg/Lane)

    Predicted band size: 56 kDa
    Observed band size: 50 kDa

    Exposure time: Lane 1-7 (left): 1 minute; Lane 1-7 (right): 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 (HA721650) at 1/5,000 dilution and competitor's antibody at 1/1,000 dilution were 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.

  • Immunohistochemical analysis of paraffin-embedded human liver tissue with Rabbit anti-LAP3 antibody (<a href="/products/HA721650" style="font-weight: bold;text-decoration: underline;">HA721650</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/HA721650" style="font-weight: bold;text-decoration: underline;">HA721650</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 liver tissue with Rabbit anti-LAP3 antibody (HA721650) 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 (HA721650) 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.

  • Western blot analysis of LAP3 on different lysates with Rabbit anti-LAP3 antibody (<a href="/products/HA721650" style="font-weight: bold;text-decoration: underline;">HA721650</a>) at 1/1,000 dilution.<br /><br />Lane 1: HeLa cell lysate (20 µg/Lane)<br />Lane 2: HepG2 cell lysate (20 µg/Lane)<br />Lane 3: HEK-293 cell lysate (20 µg/Lane)<br />Lane 4: U-87 MG cell lysate (20 µg/Lane)<br />Lane 5: K-562 cell lysate (20 µg/Lane)<br />Lane 6: A549 cell lysate (20 µg/Lane)<br />Lane 7: LO2 cell lysate (20 µg/Lane)<br />Lane 8: C6 cell lysate (20 µg/Lane)<br />Lane 9: Mouse liver tissue lysate (40 µg/Lane)<br />Lane 10: Rat liver tissue lysate (40 µg/Lane)<br /><br />Predicted band size: 56 kDa<br />Observed band size: 50 kDa<br /><br />Exposure time: 1 minute 40 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/HA721650" style="font-weight: bold;text-decoration: underline;">HA721650</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 LAP3 on different lysates with Rabbit anti-LAP3 antibody (HA721650) at 1/1,000 dilution.

    Lane 1: HeLa cell lysate (20 µg/Lane)
    Lane 2: HepG2 cell lysate (20 µg/Lane)
    Lane 3: HEK-293 cell lysate (20 µg/Lane)
    Lane 4: U-87 MG cell lysate (20 µg/Lane)
    Lane 5: K-562 cell lysate (20 µg/Lane)
    Lane 6: A549 cell lysate (20 µg/Lane)
    Lane 7: LO2 cell lysate (20 µg/Lane)
    Lane 8: C6 cell lysate (20 µg/Lane)
    Lane 9: Mouse liver tissue lysate (40 µg/Lane)
    Lane 10: Rat liver tissue lysate (40 µg/Lane)

    Predicted band size: 56 kDa
    Observed band size: 50 kDa

    Exposure time: 1 minute 40 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 (HA721650) 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 human testis tissue with Rabbit anti-LAP3 antibody (<a href="/products/HA721650" style="font-weight: bold;text-decoration: underline;">HA721650</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/HA721650" style="font-weight: bold;text-decoration: underline;">HA721650</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-LAP3 antibody (HA721650) 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 (HA721650) 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 liver tissue with Rabbit anti-LAP3 antibody (<a href="/products/HA721650" style="font-weight: bold;text-decoration: underline;">HA721650</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/HA721650" style="font-weight: bold;text-decoration: underline;">HA721650</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 liver tissue with Rabbit anti-LAP3 antibody (HA721650) 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 (HA721650) 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;">☑ Knockdown (KD)</span><br /><br />Western blot analysis of LAP3 on different lysates with Rabbit anti-LAP3 antibody (<a href="/products/HA721650" style="font-weight: bold;text-decoration: underline;">HA721650</a>) at 1/2,000 dilution.<br /><br />Lane 1: HepG2-si NT cell lysate<br />Lane 2: HepG2-si LAP3 cell lysate<br /><br />Lysates/proteins at 10 µg/Lane.<br /><br />Predicted band size: 56 kDa<br />Observed band size: 50 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/HA721650" style="font-weight: bold;text-decoration: underline;">HA721650</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 1/100,000 dilution was used for 1 hour at room temperature.

    ☑ Knockdown (KD)

    Western blot analysis of LAP3 on different lysates with Rabbit anti-LAP3 antibody (HA721650) at 1/2,000 dilution.

    Lane 1: HepG2-si NT cell lysate
    Lane 2: HepG2-si LAP3 cell lysate

    Lysates/proteins at 10 µg/Lane.

    Predicted band size: 56 kDa
    Observed band size: 50 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 (HA721650) at 1/2,000 dilution was used in 5% NFDM/TBST at 4℃ overnight. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1/100,000 dilution was used for 1 hour at room temperature.

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