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IQGAP1 Recombinant Rabbit Monoclonal Antibody [JE50-66]

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Specification

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Catalog# HA721972

IQGAP1 Recombinant Rabbit Monoclonal Antibody [JE50-66]

Application

  • WB

  • IF-Cell

  • IHC-P

  • FC

Reactivity

  • Human

  • Mouse

  • Rat

Conjugation

  • unconjugated

Overview

Product Name

IQGAP1 Recombinant Rabbit Monoclonal Antibody [JE50-66]

Antibody Type

Recombinant Rabbit monoclonal Antibody

Immunogen

Synthetic peptide within Human IQGAP1 aa 1-50 / 1,657.

Species Reactivity

Human, Mouse, Rat

Validated Applications

WB, IF-Cell, IHC-P, FC

Molecular Weight

Predicted band size:

Positive Control

HeLa cell lysate, 293T cell lysate, MDA-MB-231 cell lysate, RAW264.7 cell lysate, NIH/3T3 cell lysate, C6 cell lysate, HeLa, human liver tissue.

Conjugation

unconjugated

Clone Number

JE50-66

RRID

AB_3096790

Product Features

Form

Liquid

Concentration

1 mg/mL.(The concentration of this product may be batch-dependent)

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

  • 1:2,000-1:5,000

  • IF-Cell

  • 1:100

  • IHC-P

  • 1:4,000

  • FC

  • 1:1,000

Target

Function

This gene encodes a member of the IQGAP family. The protein contains four IQ domains, one calponin homology domain, one Ras-GAP domain and one WW domain. It interacts with components of the cytoskeleton such as the formin Dia1 (mDia1), with cell adhesion molecules (CAMs), and with several signaling molecules to regulate cell morphology and motility. For example, IQGAP1 expression is necessary for neuronal process outgrowth on the cell adhesion molecule PTPmu (PTPRM). Expression of the protein is upregulated by gene amplification in two gastric cancer cell lines and its over-expression and distinct membrane localisation is also observed in a range of tumours.

Background References

1. Liao Y et al. The Ras GTPase-activating-like protein IQGAP1 bridges Gasdermin D to the ESCRT system to promote IL-1β release via exosomes. EMBO J. 2023 Jan

2. Wei T et al. Role of IQGAP1 in Carcinogenesis. Cancers (Basel). 2021 Aug

Subcellular Location

Cell membrane, Nucleus, Cytoplasm, Apical cell membrane, Basolateral cell membrane.

UNIPROT

SwissProt: P46940 Human

SwissProt: Q9JKF1 Mouse

SwissProt: B5DFH1 Rat

Synonyms

HUMORFA01 antibody

IQ motif containing GTPase activating protein 1 antibody

IQGA1_HUMAN antibody

Iqgap1 antibody

KIAA0051 antibody

p195 antibody

Ras GTPase activating like protein 1 antibody

Ras GTPase-activating-like protein IQGAP1 antibody

RasGAP-like with IQ motifs antibody

SAR1 antibody

Images

  • Western blot analysis of IQGAP1 on different lysates with Rabbit anti-IQGAP1 antibody (<a href="/products/HA721972" style="font-weight: bold;text-decoration: underline;">HA721972</a>) at 1/2,000 dilution.<br /><br />Lane 1: HeLa cell lysate<br />Lane 2: 293T cell lysate<br />Lane 3: MDA-MB-231 cell lysate<br />Lane 4: RAW264.7 cell lysate<br />Lane 5: NIH/3T3 cell lysate<br />Lane 6: C6 cell lysate<br /><br />Lysates/proteins at 20 µg/Lane.<br /><br />Predicted band size: 189 kDa<br />Observed band size: 189 kDa<br /><br />Exposure time: 1 minute 40 second;<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/HA721972" style="font-weight: bold;text-decoration: underline;">HA721972</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 1/50,000 dilution was used for 1 hour at room temperature.

    Western blot analysis of IQGAP1 on different lysates with Rabbit anti-IQGAP1 antibody (HA721972) at 1/2,000 dilution.

    Lane 1: HeLa cell lysate
    Lane 2: 293T cell lysate
    Lane 3: MDA-MB-231 cell lysate
    Lane 4: RAW264.7 cell lysate
    Lane 5: NIH/3T3 cell lysate
    Lane 6: C6 cell lysate

    Lysates/proteins at 20 µg/Lane.

    Predicted band size: 189 kDa
    Observed band size: 189 kDa

    Exposure time: 1 minute 40 second;

    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 (HA721972) at 1/2,000 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 IQGAP1 on different lysates with Rabbit anti-IQGAP1 antibody (<a href="/products/HA721972" style="font-weight: bold;text-decoration: underline;">HA721972</a>) at 1/5,000 dilution.<br /><br />Lane 1: HAP1-parental cell lysate<br />Lane 2: HAP1-IQGAP1 KD cell lysate<br /><br />Lysates/proteins at 10 µg/Lane.<br /><br />Predicted band size: 189 kDa<br />Observed band size: 189 kDa<br /><br />Exposure time: 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/HA721972" style="font-weight: bold;text-decoration: underline;">HA721972</a>) at 1/5,000 dilution was used in <a href="/products/K1803" style="font-weight: bold;text-decoration: underline;">K1803</a> 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 IQGAP1 on different lysates with Rabbit anti-IQGAP1 antibody (HA721972) at 1/5,000 dilution.

    Lane 1: HAP1-parental cell lysate
    Lane 2: HAP1-IQGAP1 KD cell lysate

    Lysates/proteins at 10 µg/Lane.

    Predicted band size: 189 kDa
    Observed band size: 189 kDa

    Exposure time: 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 (HA721972) at 1/5,000 dilution was used in 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 IQGAP1 with Rabbit anti-IQGAP1 antibody (<a href="/products/HA721972" style="font-weight: bold;text-decoration: underline;">HA721972</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-IQGAP1 antibody (<a href="/products/HA721972" style="font-weight: bold;text-decoration: underline;">HA721972</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.

    Immunocytochemistry analysis of HeLa cells labeling IQGAP1 with Rabbit anti-IQGAP1 antibody (HA721972) 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-IQGAP1 antibody (HA721972) 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.

  • Immunohistochemical analysis of paraffin-embedded human liver tissue with Rabbit anti-IQGAP1 antibody (<a href="/products/HA721972" style="font-weight: bold;text-decoration: underline;">HA721972</a>) at 1/4,000 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/HA721972" style="font-weight: bold;text-decoration: underline;">HA721972</a>) at 1/4,000 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-IQGAP1 antibody (HA721972) at 1/4,000 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 (HA721972) at 1/4,000 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 IQGAP1.<br /><br />Cells were fixed and permeabilized. Then stained with the primary antibody (<a href="/products/HA721972" style="font-weight: bold;text-decoration: underline;">HA721972</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 IQGAP1.

    Cells were fixed and permeabilized. Then stained with the primary antibody (HA721972, 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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