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PTP1B Mouse Monoclonal Antibody [D0-C7]

Usd: 350

Specification

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Catalog# M1511-7

PTP1B Mouse Monoclonal Antibody [D0-C7]

Application

  • WB

  • IF-Cell

  • IHC-P

Reactivity

  • Human

  • Mouse

  • Rat

Conjugation

  • unconjugated

This product has been cited in peer reviewed publications, see list HERE

Overview

Product Name

PTP1B Mouse Monoclonal Antibody [D0-C7]

Antibody Type

Mouse Monoclonal Antibody

Immunogen

Recombinant protein within human PTP1B aa 1-250.

Species Reactivity

Human, Mouse, Rat

Validated Applications

WB, IF-Cell, IHC-P

Molecular Weight

Predicted band size: 50 kDa

Positive Control

A431 cell lysate, Hela cell lysate, A549 cell lysate, HepG2 cell lysate, RAW264.7 cell lysate, PC-12 cell lysate, C6 cell lysate, A431, human lung cancer tissue, mouse lung tissue.

Conjugation

unconjugated

Clone Number

D0-C7

RRID

AB_3073157

Product Features

Form

Liquid

Concentration

Storage Instructions

Shipped at 4℃. Store at +4℃ short term (1-2 weeks). It is recommended to aliquot into single-use upon delivery. Store at -20℃ long term.

Storage Buffer

1*PBS (pH7.4), 0.2% BSA, 40% Glycerol. Preservative: 0.05% Sodium Azide.

Isotype

IgG1

Purification Method

Protein A affinity purified.

Application Dilution

  • WB

  • 1:1,000-1:2,000

  • IF-Cell

  • 1:100-1:500

  • IHC-P

  • 1:200-1:1,000

Target

Function

The phosphorylation of proteins at tyrosine residues has long been recognized as an important regulatory component of signal transduction. This is a reversible process, involving both enzymes that phosphorylate proteins on tyrosine residues as well as a rapidly expanding family of protein tyrosine phosphatases. These latter enzymes bear little resemblance to either the protein serine and protein threonine phosphatases or to the acid and alkaline phosphatases. In most tissues, the major PTPase is a vanadate- and molybdate-sensitive protein. On the basis of sequence analysis, PTP1B (PTPase 1B) expressed in human placenta exhibits similarities both with the common leukocyte antigen (CD45) and with LAR, a homolog of the neural adhesion molecule (NCAM). PTP1B is synthesized as a 435 amino acid precursor protein which is cleaved to generate the active 321 amino acid enzyme.

Background References

1. Borges Bde C, et al. Protein tyrosine phosphatase-1B contributes to LPS-induced leptin resistance in male rats. Am J Physiol Endocrinol Metab 308:E40-50 (2015).

2. Krishnan N., et al. H2s-induced sulfhydration of the phosphatase PTP1B and its role in the endoplasmic reticulum stress response. Sci. Signal. 4:RA86-RA86(2011).

Sequence Similarity

Belongs to the protein-tyrosine phosphatase family. Non-receptor class 1 subfamily.

Tissue Specificity

Expressed in keratinocytes (at protein level).

Post-translational Modification

Oxidized on Cys-215; the Cys-SOH formed in response to redox signaling reacts with the alpha-amido of the following residue to form a sulfenamide cross-link, triggering a conformational change that inhibits substrate binding and activity. The active site can be restored by reduction.; Ser-50 is the major site of phosphorylation as compared to Ser-242 and Ser-243. Activated by phosphorylation at Ser-50.; S-nitrosylation of Cys-215 inactivates the enzyme activity.; Sulfhydration at Cys-215 following endoplasmic reticulum stress inactivates the enzyme activity, promoting EIF2AK3/PERK activity.

Subcellular Location

Endoplasmic reticulum membrane

UNIPROT

SwissProt: P18031 Human

SwissProt: P35821 Mouse

SwissProt: P20417 Rat

Synonyms

PTP1B antibody

Non receptor tyrosine phosphatase 1 antibody

Protein phosphotyrosylphosphatase 1B antibody

Protein tyrosine phosphatase 1B antibody

Protein tyrosine phosphatase non receptor type 1 antibody

Protein tyrosine phosphatase placental antibody

Protein-tyrosine phosphatase 1B antibody

PTN1_HUMAN antibody

PTP 1B antibody

PTP-1B antibody

Expand

Images

  • <span style="font-weight: bold;">☑ Knockdown (KD)</span><br /><br />Western blot analysis of PTP1B on different lysates with Mouse anti-PTP1B antibody (<a href="/products/M1511-7" style="font-weight: bold;text-decoration: underline;">M1511-7</a>) at 1/1,000 dilution.<br /><br />Lane 1: HAP1-parental cell lysate<br />Lane 2: HAP1-PTP1B KD cell lysate<br /><br />Lysates/proteins at 10 µg/Lane.<br /><br />Predicted band size: 50 kDa<br />Observed band size: 50 kDa<br /><br />Exposure time: 1 minute; 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/M1511-7" style="font-weight: bold;text-decoration: underline;">M1511-7</a>) at 1/1,000 dilution was used in primary antibody dilution (K1803) 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 PTP1B on different lysates with Mouse anti-PTP1B antibody (M1511-7) at 1/1,000 dilution.

    Lane 1: HAP1-parental cell lysate
    Lane 2: HAP1-PTP1B KD cell lysate

    Lysates/proteins at 10 µg/Lane.

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

    Exposure time: 1 minute; 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 (M1511-7) at 1/1,000 dilution was used in primary antibody dilution (K1803) at 4℃ overnight. Goat Anti-Mouse IgG - HRP Secondary Antibody (HA1006) at 1/50,000 dilution was used for 1 hour at room temperature.

  • Western blot analysis of PTP1B on different lysates with Mouse anti-PTP1B antibody (<a href="/products/M1511-7" style="font-weight: bold;text-decoration: underline;">M1511-7</a>) at 1/1,000 dilution.<br /><br />Lane 1: RAW264.7 cell lysate<br />Lane 2: PC-12 cell lysate<br />Lane 3: C6 cell lysate<br /><br />Lysates/proteins at 10 µg/Lane.<br /><br />Predicted band size: 50 kDa<br />Observed band size: 50 kDa<br /><br />Exposure time: 43 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/M1511-7" style="font-weight: bold;text-decoration: underline;">M1511-7</a>) at 1/1,000 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 PTP1B on different lysates with Mouse anti-PTP1B antibody (M1511-7) at 1/1,000 dilution.

    Lane 1: RAW264.7 cell lysate
    Lane 2: PC-12 cell lysate
    Lane 3: C6 cell lysate

    Lysates/proteins at 10 µg/Lane.

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

    Exposure time: 43 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 (M1511-7) at 1/1,000 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.

  • Western blot analysis of PTP1B on different lysates with Mouse anti-PTP1B antibody (<a href="/products/M1511-7" style="font-weight: bold;text-decoration: underline;">M1511-7</a>) at 1/500 dilution.<br /><br />Lane 1: A431 cell lysate<br />Lane 2: Hela cell lysate<br />Lane 3: A549 cell lysate<br />Lane 4: HepG2 cell lysate<br /><br />Lysates/proteins at 10 µg/Lane.<br /><br />Predicted band size: 50 kDa<br />Observed band size: 50 kDa<br /><br />Exposure time: 2 minutes;<br /><br />10% 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/M1511-7" style="font-weight: bold;text-decoration: underline;">M1511-7</a>) at 1/500 dilution was used in 5% NFDM/TBST at room temperature for 2 hours. Goat Anti-Mouse IgG - HRP Secondary Antibody (<a href="/products/HA1006" style="font-weight: bold;text-decoration: underline;">HA1006</a>) at 1:100,000 dilution was used for 1 hour at room temperature.

    Western blot analysis of PTP1B on different lysates with Mouse anti-PTP1B antibody (M1511-7) at 1/500 dilution.

    Lane 1: A431 cell lysate
    Lane 2: Hela cell lysate
    Lane 3: A549 cell lysate
    Lane 4: HepG2 cell lysate

    Lysates/proteins at 10 µg/Lane.

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

    Exposure time: 2 minutes;

    10% 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 (M1511-7) at 1/500 dilution was used in 5% NFDM/TBST at room temperature for 2 hours. Goat Anti-Mouse IgG - HRP Secondary Antibody (HA1006) at 1:100,000 dilution was used for 1 hour at room temperature.

  • Immunocytochemistry analysis of A431 cells labeling PTP1B with Mouse anti-PTP1B antibody (<a href="/products/M1511-7" style="font-weight: bold;text-decoration: underline;">M1511-7</a>) at 1/100 dilution.<br /><br />Cells were fixed in 4% paraformaldehyde for 15 minutes at room temperature, permeabilized with 0.1% Triton X-100 in PBS for 15 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 Mouse anti-PTP1B antibody (<a href="/products/M1511-7" style="font-weight: bold;text-decoration: underline;">M1511-7</a>) at 1/100 dilution in 1% BSA in PBST 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. 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/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 A431 cells labeling PTP1B with Mouse anti-PTP1B antibody (M1511-7) at 1/100 dilution.

    Cells were fixed in 4% paraformaldehyde for 15 minutes at room temperature, permeabilized with 0.1% Triton X-100 in PBS for 15 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 Mouse anti-PTP1B antibody (M1511-7) at 1/100 dilution in 1% BSA in PBST overnight at 4 ℃. Goat Anti-Mouse IgG H&L (iFluor™ 488, HA1125) 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 (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.

  • Immunohistochemical analysis of paraffin-embedded human lung cancer tissue with Mouse anti-PTP1B antibody (<a href="/products/M1511-7" style="font-weight: bold;text-decoration: underline;">M1511-7</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/M1511-7" style="font-weight: bold;text-decoration: underline;">M1511-7</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 lung cancer tissue with Mouse anti-PTP1B antibody (M1511-7) 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 (M1511-7) 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 lung tissue with Mouse anti-PTP1B antibody (<a href="/products/M1511-7" style="font-weight: bold;text-decoration: underline;">M1511-7</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/M1511-7" style="font-weight: bold;text-decoration: underline;">M1511-7</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 lung tissue with Mouse anti-PTP1B antibody (M1511-7) 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 (M1511-7) 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.

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Citation

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