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HMGA1 Recombinant Rabbit Monoclonal Antibody [JE58-17]

Usd: 385

Specification

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Catalog# HA722280

HMGA1 Recombinant Rabbit Monoclonal Antibody [JE58-17]

Application

  • WB

  • IF-Cell

  • FC

Reactivity

  • Human

Conjugation

  • unconjugated

Overview

Product Name

HMGA1 Recombinant Rabbit Monoclonal Antibody [JE58-17]

Antibody Type

Recombinant Rabbit monoclonal Antibody

Immunogen

Recombinant protein within Human HMGA1 aa 1-80 / 107.

Species Reactivity

Human

Validated Applications

WB, IF-Cell, FC

Target Molecular Weight

Predicted band size: 12 kDa

Positive Control

A431 cell lysate, HCT 116 cell lysate, HepG2 cell lysate, MDA-MB-231 cell lysate, HCT 116.

Conjugation

unconjugated

Clone Number

JE58-17

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

  • 1:1,000

  • IF-Cell

  • 1:200

  • FC

  • 1:1,000

Target

Function

High-mobility group protein HMG-I/HMG-Y is a protein that in humans is encoded by the HMGA1 gene. This gene encodes a non-histone chromatin protein involved in many cellular processes, including regulation of inducible gene transcription, DNA replication, heterochromatin organization, integration of retroviruses into chromosomes, and the metastatic progression of cancer cells. HMGA1 proteins are quite small (~10-12 kDa) and basic molecules, and consist of three AT-hooks with the RGRP (Arg-Gly-Arg-Pro) core motif, a novel cross-linking domain located between the second and third AT-hook, and a C-terminal acidic tail characteristic for the HMG family comprising HMGA, HMGB and HMGN proteins.

Background References

1. Wang Y. et. al. HMGA1 in cancer: Cancer classification by location. J Cell Mol Med. 2019 Apr

2. Resar L. et. al. Lessons from the Crypt: HMGA1-Amping up Wnt for Stem Cells and Tumor Progression. Cancer Res. 2018 Apr

Subcellular Location

Nucleus, Chromosome.

UNIPROT

SwissProt: P17096 Human

Synonyms

High mobility group (nonhistone chromosomal) protein isoforms I and Y antibody

High mobility group AT hook 1 antibody

High mobility group AT-hook protein 1 antibody

High mobility group protein 1 antibody

High mobility group protein A1 antibody

High mobility group protein HMG-I/HMG-Y antibody

High mobility group protein Ia antibody

High mobility group protein R antibody

HMG R antibody

HMG-I(Y) antibody

Expand

Images

  • Western blot analysis of HMGA1 on different lysates with Rabbit anti-HMGA1 antibody (<a href="/products/HA722280" style="font-weight: bold;text-decoration: underline;">HA722280</a>) at 1/1,000 dilution.<br /><br />Lane 1: A431 cell lysate<br />Lane 2: HCT 116 cell lysate<br />Lane 3: HepG2 cell lysate<br />Lane 4: MDA-MB-231 cell lysate<br /><br />Lysates/proteins at 20 µg/Lane.<br /><br />Predicted band size: 12 kDa<br />Observed band size: 17 kDa<br /><br />Exposure time: 1 minute 18 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/HA722280" style="font-weight: bold;text-decoration: underline;">HA722280</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.

    Western blot analysis of HMGA1 on different lysates with Rabbit anti-HMGA1 antibody (HA722280) at 1/1,000 dilution.

    Lane 1: A431 cell lysate
    Lane 2: HCT 116 cell lysate
    Lane 3: HepG2 cell lysate
    Lane 4: MDA-MB-231 cell lysate

    Lysates/proteins at 20 µg/Lane.

    Predicted band size: 12 kDa
    Observed band size: 17 kDa

    Exposure time: 1 minute 18 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 (HA722280) 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.

  • Immunocytochemistry analysis of HCT 116 cells labeling HMGA1 with Rabbit anti-HMGA1 antibody (<a href="/products/HA722280" style="font-weight: bold;text-decoration: underline;">HA722280</a>) at 1/200 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-HMGA1 antibody (<a href="/products/HA722280" style="font-weight: bold;text-decoration: underline;">HA722280</a>) at 1/200 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 HCT 116 cells labeling HMGA1 with Rabbit anti-HMGA1 antibody (HA722280) at 1/200 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-HMGA1 antibody (HA722280) at 1/200 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 HCT 116 cells labeling HMGA1.<br /><br />Cells were fixed and permeabilized. Then stained with the primary antibody (<a href="/products/HA722280" style="font-weight: bold;text-decoration: underline;">HA722280</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 HCT 116 cells labeling HMGA1.

    Cells were fixed and permeabilized. Then stained with the primary antibody (HA722280, 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).

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