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Lin28A Recombinant Rabbit Monoclonal Antibody [JE60-36]

Usd: 385

Specification

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Catalog# HA722899

Lin28A Recombinant Rabbit Monoclonal Antibody [JE60-36]

Application

  • WB

  • IF-Cell

  • FC

  • IP

Reactivity

  • Human

Conjugation

  • unconjugated

Overview

Product Name

Lin28A Recombinant Rabbit Monoclonal Antibody [JE60-36]

Antibody Type

Recombinant Rabbit monoclonal Antibody

Immunogen

Synthetic peptide within human Lin28A aa 160-209 / 209.

Species Reactivity

Human

Validated Applications

WB, IF-Cell, FC, IP

Target Molecular Weight

Predicted band size: 23 kDa

Positive Control

NCCIT cell lysate, JAR cell lysate, NCCIT.

Conjugation

unconjugated

Clone Number

JE60-36

Reactivity Data

WB IF-Cell FC IP
Human
Tested Tested
Tested Tested
Tested Tested
Tested Tested
Mouse
Not Recommended
Not Recommended
Rat
Not Recommended

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

PBS (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

  • 1:500

  • FC

  • 1:1,000

  • IP

  • 1-2μg/sample

Target

Function

LIN28A is conserved, developmentally regulated RNA binding proteins that inhibit the processing and maturation of the let-7 family of miRNAs. LIN28A is localized to the periendoplasmic reticulum (ER) area and inhibits translation of mRNAs that are destined for the ER, reducing the synthesis of transmembrane proteins, ER or Golgi lumen proteins, and secretory proteins. Overexpression of LIN28A, in conjunction with Oct-4, Sox2, and Nanog, can reprogram human fibroblasts to pluripotent, ES-like cells.

Background References

1. Oyejobi GK et al. Regulating Protein-RNA Interactions: Advances in Targeting the LIN28/Let-7 Pathway. Int J Mol Sci. 2024 Mar.

2. Zhou F et al. Circular RNA circ_0096041 promotes osteosarcoma cell proliferation and migration via sponging miR-556-5p and regulating LIN28A expression. Cell Mol Biol (Noisy-le-grand). 2024 Feb.

Subcellular Location

Cytoplasm, Nucleus.

UNIPROT

SwissProt: Q9H9Z2 Human

Synonyms

CSDD1 antibody

FLJ12457 antibody

LIN 28 antibody

Lin 28 homolog A (C. elegans) antibody

Lin-28A antibody

LIN28 antibody

LIN28A antibody

LN28A_HUMAN antibody

Protein lin-28 homolog A antibody

ZCCHC1 antibody

Expand

Images

  • <span style="font-weight: bold;">☑ Relative expression (RE)</span><br /><br />Western blot analysis of Lin28A on different lysates with Rabbit anti-Lin28A antibody (<a href="/products/HA722899" style="font-weight: bold;text-decoration: underline;">HA722899</a>) at 1/1,000 dilution.<br /><br />Lane 1: NCCIT cell lysate<br />Lane 2: JAR cell lysate<br />Lane 3: HeLa cell lysate (negative)<br /><br />Lysates/proteins at 20 µg/Lane.<br /><br />Predicted band size: 23 kDa<br />Observed band size: 29 kDa<br /><br />Exposure time: 4 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/HA722899" style="font-weight: bold;text-decoration: underline;">HA722899</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 Lin28A on different lysates with Rabbit anti-Lin28A antibody (HA722899) at 1/1,000 dilution.

    Lane 1: NCCIT cell lysate
    Lane 2: JAR cell lysate
    Lane 3: HeLa cell lysate (negative)

    Lysates/proteins at 20 µg/Lane.

    Predicted band size: 23 kDa
    Observed band size: 29 kDa

    Exposure time: 4 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 (HA722899) 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;">☑ Relative expression (RE)</span><br /><br />Immunocytochemistry analysis of NCCIT (positive) and HeLa (negative) labeling Lin28A with Rabbit anti-Lin28A antibody (<a href="/products/HA722899" style="font-weight: bold;text-decoration: underline;">HA722899</a>) at 1/500 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 Rabbit anti-Lin28A antibody (<a href="/products/HA722899" style="font-weight: bold;text-decoration: underline;">HA722899</a>) at 1/500 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.

    ☑ Relative expression (RE)

    Immunocytochemistry analysis of NCCIT (positive) and HeLa (negative) labeling Lin28A with Rabbit anti-Lin28A antibody (HA722899) at 1/500 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 Rabbit anti-Lin28A antibody (HA722899) at 1/500 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 NCCIT cells labeling Lin28A.<br /><br />Cells were fixed and permeabilized. Then stained with the primary antibody (<a href="/products/HA722899" style="font-weight: bold;text-decoration: underline;">HA722899</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 NCCIT cells labeling Lin28A.

    Cells were fixed and permeabilized. Then stained with the primary antibody (HA722899, 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).

  • Lin28A was immunoprecipitated from 0.2 mg JAR cell lysate with <a href="/products/HA722899" style="font-weight: bold;text-decoration: underline;">HA722899</a> at 2 µg/10 µl beads. Western blot was performed from the immunoprecipitate using <a href="/products/HA722899" style="font-weight: bold;text-decoration: underline;">HA722899</a> at 1/1,000 dilution. Anti-Rabbit IgG for IP, AlpSdAbs&reg; VHH(HRP) (<a href="/products/025-100-005" style="font-weight: bold;text-decoration: underline;">025-100-005</a>) at 1/5,000 dilution was used for 1 hour at room temperature.<br /><br />Lane 1: JAR cell lysate (input)<br />Lane 2: <a href="/products/HA722899" style="font-weight: bold;text-decoration: underline;">HA722899</a> IP in JAR cell lysate<br />Lane 3: Rabbit IgG instead of <a href="/products/HA722899" style="font-weight: bold;text-decoration: underline;">HA722899</a> in JAR cell lysate<br /><br />Blocking/Dilution buffer: 5% NFDM/TBST<br />Exposure time: 6 seconds; ECL: <a href="/products/K1801" style="font-weight: bold;text-decoration: underline;">K1801</a>

    Lin28A was immunoprecipitated from 0.2 mg JAR cell lysate with HA722899 at 2 µg/10 µl beads. Western blot was performed from the immunoprecipitate using HA722899 at 1/1,000 dilution. Anti-Rabbit IgG for IP, AlpSdAbs® VHH(HRP) (025-100-005) at 1/5,000 dilution was used for 1 hour at room temperature.

    Lane 1: JAR cell lysate (input)
    Lane 2: HA722899 IP in JAR cell lysate
    Lane 3: Rabbit IgG instead of HA722899 in JAR cell lysate

    Blocking/Dilution buffer: 5% NFDM/TBST
    Exposure time: 6 seconds; ECL: K1801

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