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Galectin 3 Recombinant Rabbit Monoclonal Antibody [JF39-10]

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Specification

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Catalog# ET1702-48

Galectin 3 Recombinant Rabbit Monoclonal Antibody [JF39-10]

Application

  • WB

  • IF-Cell

  • IF-Tissue

  • FC

  • IHC-P

Reactivity

  • Human

  • Mouse

  • Rat

Conjugation

  • unconjugated

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

Overview

Product Name

Galectin 3 Recombinant Rabbit Monoclonal Antibody [JF39-10]

Antibody Type

Recombinant Rabbit monoclonal Antibody

Immunogen

Synthetic peptide within Human Galectin 3 aa 1-49 / 250.

Species Reactivity

Human, Mouse, Rat

Validated Applications

WB, IF-Cell, IF-Tissue, FC, IHC-P

Molecular Weight

Predicted band size: 26 kDa

Positive Control

HeLa cell lysate, RAW264.7 cell lysate, Rat colon tissue lysate, Rat brain tissue lysate, PC-12 cell lysate, HeLa, human thyroid carcinoma tissue, human colon tissue, rat bladder tissue.

Conjugation

unconjugated

Clone Number

JF39-10

RRID

AB_3070310

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

Isotype

IgG

Purification Method

Protein A affinity purified.

Application Dilution

  • WB

  • 1:500-1:2,000

  • IF-Cell

  • 1:50-1:200

  • IF-Tissue

  • 1:50-1:200

  • FC

  • 1:1,000

  • IHC-P

  • 1:200-1:1,000

Target

Function

Galectins are a family of soluble b-galactoside-binding animal lectins that modulate cell-to-cell adhesion and cell-to-extracellular matrix (ECM) inter- actions and play a role in tumor progression, pre-mRNA splicing and apop-tosis. The galectin-3 protein, also known as Mac-2, hMac-2, GALBP, CBP35 or LGALS3, contains a single carbohydrate binding domain, which binds galactose-containing glycoconjugates. Galectin-3 is expressed in colonic and intestinal epithelium, inflammatory macrophages, papillary and follicular carcinomas, neoplastic astrocytes and some B and T lymphocytes. Upregulated expression of galectin-3 is involved in cancer progression and metastasis. Galectin-3 mediates the endocytosis of β1 Integrins in a lactose-dependent manner and is associated with thyroid malignancy and Crohn's disease. It may also be used as a marker for diagnosing cases involving Hurthle cell adenomas and carcinomas.

Background References

1. Qiao L et al. Gene silencing of galectin-3 changes the biological behavior of Eca109 human esophageal cancer cells. Mol Med Rep 13:160-6 (2016).

2. Lin YW et al. RIP140 contributes to foam cell formation and atherosclerosis by regulating cholesterol homeostasis in macrophages. J Mol Cell Cardiol 79:287-94 (2015).

Tissue Specificity

A major expression is found in the colonic epithelium. It is also abundant in the activated macrophages. Expressed in fetal membranes.

Subcellular Location

Nucleus, Secreted, Cytoplasm.

UNIPROT

SwissProt: P17931 Human

SwissProt: P16110 Mouse

SwissProt: P08699 Rat

Synonyms

35 kDa lectin antibody

Carbohydrate binding protein 35 antibody

Carbohydrate-binding protein 35 antibody

CBP 35 antibody

CBP35 antibody

Gal-3 antibody

GAL3 antibody

Galactose-specific lectin 3 antibody

Galactoside-binding protein antibody

GALBP antibody

Expand

Images

  • <span style="font-weight: bold;">☑ Relative expression (RE)</span><br /><br />Western blot analysis of Galectin 3 on different lysates with Rabbit anti-Galectin 3 antibody (<a href="/products/ET1702-48" style="font-weight: bold;text-decoration: underline;">ET1702-48</a>) at 1/1,000 dilution.<br /><br />Lane 1: HeLa cell lysate (20 µg/Lane)<br />Lane 2: RAW264.7 cell lysate (20 µg/Lane)<br />Lane 3: Rat colon tissue lysate (40 µg/Lane)<br />Lane 4: Rat brain tissue lysate (low expression) (40 µg/Lane)<br /><br />Predicted band size: 26 kDa<br />Observed band size: 30 kDa<br /><br />Exposure time: 10 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/ET1702-48" style="font-weight: bold;text-decoration: underline;">ET1702-48</a>) at 1/1,000 dilution was used in primary antibody dilution (K1803) 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.

    ☑ Relative expression (RE)

    Western blot analysis of Galectin 3 on different lysates with Rabbit anti-Galectin 3 antibody (ET1702-48) at 1/1,000 dilution.

    Lane 1: HeLa cell lysate (20 µg/Lane)
    Lane 2: RAW264.7 cell lysate (20 µg/Lane)
    Lane 3: Rat colon tissue lysate (40 µg/Lane)
    Lane 4: Rat brain tissue lysate (low expression) (40 µg/Lane)

    Predicted band size: 26 kDa
    Observed band size: 30 kDa

    Exposure time: 10 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 (ET1702-48) at 1/1,000 dilution was used in primary antibody dilution (K1803) at 4℃ overnight. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1/50,000 dilution was used for 1 hour at room temperature.

  • <span style="font-weight: bold;">☑ Relative expression (RE)</span><br /><br />Western blot analysis of Galectin 3 on different lysates with Rabbit anti-Galectin 3 antibody (<a href="/products/ET1702-48" style="font-weight: bold;text-decoration: underline;">ET1702-48</a>) at 1/1,000 dilution.<br /><br />Lane 1: HeLa cell lysate<br />Lane 2: LNCaP cell lysate (negative)<br />Lane 3: PC-12 cell lysate<br /><br />Lysates/proteins at 20 µg/Lane.<br /><br />Predicted band size: 26 kDa<br />Observed band size: 30 kDa<br /><br />Exposure time: 10 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/ET1702-48" style="font-weight: bold;text-decoration: underline;">ET1702-48</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.

    ☑ Relative expression (RE)

    Western blot analysis of Galectin 3 on different lysates with Rabbit anti-Galectin 3 antibody (ET1702-48) at 1/1,000 dilution.

    Lane 1: HeLa cell lysate
    Lane 2: LNCaP cell lysate (negative)
    Lane 3: PC-12 cell lysate

    Lysates/proteins at 20 µg/Lane.

    Predicted band size: 26 kDa
    Observed band size: 30 kDa

    Exposure time: 10 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 (ET1702-48) 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.

  • <span style="font-weight: bold;">☑ Knockdown (KD)</span><br /><br />Western blot analysis of Galectin 3 on different lysates with Rabbit anti-Galectin 3 antibody (<a href="/products/ET1702-48" style="font-weight: bold;text-decoration: underline;">ET1702-48</a>) at 1/1,000 dilution.<br /><br />Lane 1: HAP1-parental cell lysate<br />Lane 2: HAP1-Galectin 3 KD cell lysate<br /><br />Lysates/proteins at 10 µg/Lane.<br /><br />Predicted band size: 26 kDa<br />Observed band size: 30 kDa<br /><br />Exposure time: 2 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/ET1702-48" style="font-weight: bold;text-decoration: underline;">ET1702-48</a>) at 1/1,000 dilution was used in primary antibody dilution (K1803) 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.

    ☑ Knockdown (KD)

    Western blot analysis of Galectin 3 on different lysates with Rabbit anti-Galectin 3 antibody (ET1702-48) at 1/1,000 dilution.

    Lane 1: HAP1-parental cell lysate
    Lane 2: HAP1-Galectin 3 KD cell lysate

    Lysates/proteins at 10 µg/Lane.

    Predicted band size: 26 kDa
    Observed band size: 30 kDa

    Exposure time: 2 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 (ET1702-48) at 1/1,000 dilution was used in primary antibody dilution (K1803) 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 Galectin 3 with Rabbit anti-Galectin 3 antibody (<a href="/products/ET1702-48" style="font-weight: bold;text-decoration: underline;">ET1702-48</a>) at 1/100 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-Galectin 3 antibody (<a href="/products/ET1702-48" style="font-weight: bold;text-decoration: underline;">ET1702-48</a>) at 1/100 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 Galectin 3 with Rabbit anti-Galectin 3 antibody (ET1702-48) at 1/100 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-Galectin 3 antibody (ET1702-48) at 1/100 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.

  • Flow cytometric analysis of HeLa cells labeling Galectin 3.<br /><br />Cells were fixed and permeabilized. Then stained with the primary antibody (<a href="/products/ET1702-48" style="font-weight: bold;text-decoration: underline;">ET1702-48</a>, 1/1,000) (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 Galectin 3.

    Cells were fixed and permeabilized. Then stained with the primary antibody (ET1702-48, 1/1,000) (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).

  • Immunohistochemical analysis of paraffin-embedded human thyroid carcinoma tissue with Rabbit anti-Galectin 3 antibody (<a href="/products/ET1702-48" style="font-weight: bold;text-decoration: underline;">ET1702-48</a>) at 1/200 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/ET1702-48" style="font-weight: bold;text-decoration: underline;">ET1702-48</a>) at 1/200 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 thyroid carcinoma tissue with Rabbit anti-Galectin 3 antibody (ET1702-48) at 1/200 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 (ET1702-48) at 1/200 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 colon tissue with Rabbit anti-Galectin 3 antibody (<a href="/products/ET1702-48" style="font-weight: bold;text-decoration: underline;">ET1702-48</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/ET1702-48" style="font-weight: bold;text-decoration: underline;">ET1702-48</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 colon tissue with Rabbit anti-Galectin 3 antibody (ET1702-48) 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 (ET1702-48) 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-Galectin 3 antibody (<a href="/products/ET1702-48" style="font-weight: bold;text-decoration: underline;">ET1702-48</a>) at 1/200 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/ET1702-48" style="font-weight: bold;text-decoration: underline;">ET1702-48</a>) at 1/200 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-Galectin 3 antibody (ET1702-48) at 1/200 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 (ET1702-48) at 1/200 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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