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ASS1 Recombinant Mouse Monoclonal Antibody [11F1-RA] - BSA and Azide free

Usd: 649

Specification

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Catalog# HA610025

ASS1 Recombinant Mouse Monoclonal Antibody [11F1-RA] - BSA and Azide free

Application

  • WB

  • IHC-P

  • IF-Cell

Reactivity

  • Human

  • Mouse

  • Rat

With BSA and Azide
Conjugation

  • unconjugated

Overview

Product Name

ASS1 Recombinant Mouse Monoclonal Antibody [11F1-RA] - BSA and Azide free

Antibody Type

Recombinant Mouse Monoclonal Antibody

Immunogen

Recombinant protein within Human ASS1 aa 1-412 / 412.

Species Reactivity

Human, Mouse, Rat

Validated Applications

WB, IHC-P, IF-Cell

Molecular Weight

Predicted band size: 47 kDa

Positive Control

HepG2 cell lysate, Human liver tissue lysate, Human kidney tissue lysate, Mouse liver tissue lysate, Mouse kidney tissue lysate, Rat liver tissue lysate, Rat kidney tissue lysate, rat kidney tissue, human kidney tissue, MCF-7.

Conjugation

unconjugated

Clone Number

11F1-RA

Product Features

Form

Liquid

Concentration

1mg/ml

Storage Instructions

Store at +4℃ after thawing. Aliquot store at -20℃. Avoid repeated freeze / thaw cycles.

Storage Buffer

1*PBS (pH7.4).

Isotype

IgG2b

Purification Method

Protein A affinity purified.

Application Dilution

  • WB

  • 1:5,000-1:20,000

  • IHC-P

  • 1:1,000

  • IF-Cell

  • 1:200

Target

Function

Argininosuccinate synthetase is an enzyme that in humans is encoded by the ASS1 gene. The protein encoded by this gene catalyzes the penultimate step of the arginine biosynthetic pathway. There are approximately 10 to 14 copies of this gene including the pseudogenes scattered across the human genome, among which the one located on chromosome 9 appears to be the only functional gene for argininosuccinate synthetase. Two transcript variants encoding the same protein have been found for this gene. Mutations in the chromosome 9 copy of ASS cause citrullinemia. 40% to 90%[8] of bladder cancers are deficient in argininosuccinate synthetase.

Background References

1. Carritt B, Goldfarb PS, Hooper ML, Slack C (Jun 1977). "Chromosome assignment of a human gene for argininosuccinate synthetase expression in Chinese hamsterxhuman somatic cell hybrids". Exp Cell Res. 106 (1): 71–8.

2. Beaudet AL, O\'Brien WE, Bock HG, Freytag SO, Su TS (Mar 1986). "The human argininosuccinate synthetase locus and citrullinemia". Adv Hum Genet. 15: 161–96, 291–2.

Subcellular Location

Cytoplasm

UNIPROT

SwissProt: P00966 Human

SwissProt: P16460 Mouse

SwissProt: P09034 Rat

Synonyms

Argininosuccinate synthase 1 antibody

Argininosuccinate synthase antibody

Argininosuccinate synthetase 1 antibody

ASS antibody

Ass-1 antibody

ass1 antibody

ASSA antibody

ASSY_HUMAN antibody

Citrulline aspartate ligase antibody

Citrulline--aspartate ligase antibody

Expand

Images

  • Western blot analysis of ASS1 on different lysates with Mouse anti-ASS1 antibody (<a href="/products/HA610025" style="font-weight: bold;text-decoration: underline;">HA610025</a>) at 1/5,000 dilution.<br /><br />Lane 1: HepG2 cell lysate (15 µg/Lane)<br />Lane 2: Human liver tissue lysate (30 µg/Lane)<br />Lane 3: Human kidney tissue lysate (30 µg/Lane)<br />Lane 4: Mouse liver tissue lysate (30 µg/Lane)<br />Lane 5: Mouse kidney tissue lysate (30 µg/Lane)<br />Lane 6: Rat liver tissue lysate (30 µg/Lane)<br />Lane 7: Rat kidney tissue lysate (30 µg/Lane)<br /><br />Predicted band size: 47 kDa<br />Observed band size: 45 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/HA610025" style="font-weight: bold;text-decoration: underline;">HA610025</a>) at 1/5,000 dilution was used in 5% NFDM/TBST at 4℃ overnight. Goat Anti-Mouse IgG - HRP Secondary Antibody (<a href="/products/HA1006" style="font-weight: bold;text-decoration: underline;">HA1006</a>) at 1/50,000 dilution was used for 1 hour at room temperature.

    Western blot analysis of ASS1 on different lysates with Mouse anti-ASS1 antibody (HA610025) at 1/5,000 dilution.

    Lane 1: HepG2 cell lysate (15 µg/Lane)
    Lane 2: Human liver tissue lysate (30 µg/Lane)
    Lane 3: Human kidney tissue lysate (30 µg/Lane)
    Lane 4: Mouse liver tissue lysate (30 µg/Lane)
    Lane 5: Mouse kidney tissue lysate (30 µg/Lane)
    Lane 6: Rat liver tissue lysate (30 µg/Lane)
    Lane 7: Rat kidney tissue lysate (30 µg/Lane)

    Predicted band size: 47 kDa
    Observed band size: 45 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 (HA610025) at 1/5,000 dilution was used in 5% NFDM/TBST at 4℃ overnight. Goat Anti-Mouse IgG - HRP Secondary Antibody (HA1006) at 1/50,000 dilution was used for 1 hour at room temperature.

  • <span style="font-weight: bold;">☑ Knockdown (KD)</span><br /><br />Western blot analysis of ASS1 on different lysates with Mouse anti-ASS1 antibody (<a href="/products/HA610025" style="font-weight: bold;text-decoration: underline;">HA610025</a>) at 1/2,000 dilution.<br /><br />Lane 1: A549-WT cell lysate<br />Lane 2: A549-KD ASS1 cell lysate<br /><br />Lysates/proteins at 10 µg/Lane.<br /><br />Predicted band size: 47 kDa<br />Observed band size: 45 kDa<br /><br />Exposure time: 3 minutes; ECL: K1802;<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/HA610025" style="font-weight: bold;text-decoration: underline;">HA610025</a>) at 1/2,000 dilution was used in 5% NFDM/TBST at 4℃ overnight. Goat Anti-Mouse IgG - HRP Secondary Antibody (<a href="/products/HA1006" style="font-weight: bold;text-decoration: underline;">HA1006</a>) at 1/50,000 dilution was used for 1 hour at room temperature.

    ☑ Knockdown (KD)

    Western blot analysis of ASS1 on different lysates with Mouse anti-ASS1 antibody (HA610025) at 1/2,000 dilution.

    Lane 1: A549-WT cell lysate
    Lane 2: A549-KD ASS1 cell lysate

    Lysates/proteins at 10 µg/Lane.

    Predicted band size: 47 kDa
    Observed band size: 45 kDa

    Exposure time: 3 minutes; ECL: K1802;

    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 (HA610025) at 1/2,000 dilution was used in 5% NFDM/TBST at 4℃ overnight. Goat Anti-Mouse IgG - HRP Secondary Antibody (HA1006) at 1/50,000 dilution was used for 1 hour at room temperature.

  • Immunohistochemical analysis of paraffin-embedded rat kidney tissue with Mouse anti-ASS1 antibody (<a href="/products/HA610025" style="font-weight: bold;text-decoration: underline;">HA610025</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/HA610025" style="font-weight: bold;text-decoration: underline;">HA610025</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 kidney tissue with Mouse anti-ASS1 antibody (HA610025) 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 (HA610025) 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 kidney tissue with Mouse anti-ASS1 antibody (<a href="/products/HA610025" style="font-weight: bold;text-decoration: underline;">HA610025</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/HA610025" style="font-weight: bold;text-decoration: underline;">HA610025</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 kidney tissue with Mouse anti-ASS1 antibody (HA610025) 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 (HA610025) 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 MCF-7 cells labeling ASS1 with Mouse anti-ASS1 antibody (<a href="/products/HA610025" style="font-weight: bold;text-decoration: underline;">HA610025</a>) at 1/200 dilution.<br /><br />Cells were fixed in 4% paraformaldehyde for 30 minutes, 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 Mouse anti-ASS1 antibody (<a href="/products/HA610025" style="font-weight: bold;text-decoration: underline;">HA610025</a>) at 1/200 dilution in 2% negative goat serum overnight at 4 ℃. Goat Anti-Mouse IgG H&L (iFluor&trade; 488, <a href="/products/HA1125" style="font-weight: bold;text-decoration: underline;">HA1125</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/ET1602-4" style="font-weight: bold;text-decoration: underline;">ET1602-4</a>, red) was stained at 1/100 dilution overnight at +4℃. Goat Anti-Rabbit IgG H&L (iFluor&trade; 594, <a href="/products/HA1122" style="font-weight: bold;text-decoration: underline;">HA1122</a>) were used as the secondary antibody at 1/1,000 dilution.

    Immunocytochemistry analysis of MCF-7 cells labeling ASS1 with Mouse anti-ASS1 antibody (HA610025) at 1/200 dilution.

    Cells were fixed in 4% paraformaldehyde for 30 minutes, 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 Mouse anti-ASS1 antibody (HA610025) at 1/200 dilution in 2% negative goat serum overnight at 4 ℃. Goat Anti-Mouse IgG H&L (iFluor™ 488, HA1125) was used as the secondary antibody at 1/1,000 dilution. Nuclear DNA was labelled in blue with DAPI.

    beta Tubulin (ET1602-4, red) was stained at 1/100 dilution overnight at +4℃. Goat Anti-Rabbit IgG H&L (iFluor™ 594, HA1122) were used as the secondary antibody at 1/1,000 dilution.

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