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APP/beta Amyloid Recombinant Rabbit Monoclonal Antibody [PSH04-90] - BSA and Azide free

Usd: 649

Specification

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Catalog# HA750977

APP/beta Amyloid Recombinant Rabbit Monoclonal Antibody [PSH04-90] - BSA and Azide free

Application

  • WB

  • IF-Cell

  • IHC-P

Reactivity

  • Human

  • Mouse

With BSA and Azide
Conjugation

  • unconjugated

Overview

Product Name

APP/beta Amyloid Recombinant Rabbit Monoclonal Antibody [PSH04-90] - BSA and Azide free

Antibody Type

Recombinant Rabbit monoclonal Antibody

Immunogen

Synthetic peptide within C-terminal human Amyloid Beta.

Species Reactivity

Human, Mouse

Validated Applications

WB, IF-Cell, IHC-P

Molecular Weight

Predicted band size: 87 kDa

Positive Control

HeLa cell lysate, HeLa treated with 100ng/mL Nocodazole for 17 hours cell lysate, SH-SY5Y cell lysate, AD mouse brain tissue lysate, mouse brain tissue lysate, HeLa, AD mouse brain tissue, mouse brain tissue.

Conjugation

unconjugated

Clone Number

PSH04-90

Product Features

Form

Liquid

Concentration

1mg/ml

Storage Instructions

Store at +4℃ after thawing. Aliquot store at -20℃. Avoid repeated freeze / thaw cycles.

Storage Buffer

1*PBS (pH7.4).

Isotype

IgG

Purification Method

Immunogen affinity purified.

Application Dilution

  • WB

  • 1:1,000

  • IF-Cell

  • 1:100

  • IHC-P

  • 1:200-1:1,000

Target

Function

Amyloid beta precursor protein (APP) is a 100-140 kDa transmembrane glycoprotein existing as several isoforms. The amino acid sequence of APP contains the amyloid domain (A-beta), which can be released by a two-step proteolytic cleavage. It is expressed in all fetal tissues examined with highest levels in brain, kidney, heart and spleen. APP acts as a cell surface receptor and performs physiological functions on the surface of neurons relevant to neurite growth, neuronal adhesion and axonogenesis. It is also involved in cell mobility and transcription regulation through protein-protein interactions.

Background References

1. Cho Y et al. Physiology and pharmacology of amyloid precursor protein. Pharmacol Ther. 2022 Jul

2. Strope TA et al. Amyloid precursor protein and mitochondria. Curr Opin Neurobiol. 2023 Feb

Subcellular Location

Cell membrane, Membrane, Perikaryon, Cell projection, growth cone, Membrane, clathrin-coated pit, Early endosome, Cytoplasmic vesicle.

UNIPROT

SwissProt: P05067 Human

SwissProt: P12023 Mouse

Synonyms

A4 amyloid protein antibody

A4_HUMAN antibody

AAA antibody

ABETA antibody

ABPP antibody

AD1 antibody

AICD-50 antibody

AICD-57 antibody

AICD-59 antibody

AID(50) antibody

Expand

Images

  • <span style="font-weight: bold;">☑ Cell treatment (CT)</span><br /><br />Western blot analysis of APP/beta Amyloid on different lysates with Rabbit anti-APP/beta Amyloid antibody (<a href="/products/HA750977" style="font-weight: bold;text-decoration: underline;">HA750977</a>) at 1/1,000 dilution.<br /><br />Lane 1: HeLa cell lysate<br />Lane 2: HeLa treated with 100ng/mL Nocodazole for 17 hours cell lysate<br />Lane 3: SH-SY5Y cell lysate<br /><br />Lysates/proteins at 20 µg/Lane.<br /><br />Predicted band size: 87 kDa<br />Observed band size: 87-120 kDa<br /><br />Exposure time: 1 minute 2 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/HA750977" style="font-weight: bold;text-decoration: underline;">HA750977</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.

    ☑ Cell treatment (CT)

    Western blot analysis of APP/beta Amyloid on different lysates with Rabbit anti-APP/beta Amyloid antibody (HA750977) at 1/1,000 dilution.

    Lane 1: HeLa cell lysate
    Lane 2: HeLa treated with 100ng/mL Nocodazole for 17 hours cell lysate
    Lane 3: SH-SY5Y cell lysate

    Lysates/proteins at 20 µg/Lane.

    Predicted band size: 87 kDa
    Observed band size: 87-120 kDa

    Exposure time: 1 minute 2 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 (HA750977) 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 />Western blot analysis of APP/beta Amyloid on different lysates with Rabbit anti-APP/beta Amyloid antibody (<a href="/products/HA750977" style="font-weight: bold;text-decoration: underline;">HA750977</a>) at 1/1,000 dilution.<br /><br />Lane 1: AD Mouse brain tissue lysate<br />Lane 2: Mouse brain tissue lysate (low expression)<br /><br />Lysates/proteins at 20 µg/Lane.<br /><br />Predicted band size: 87 kDa<br />Observed band size: 100 kDa<br /><br />Exposure time: 3 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/HA750977" style="font-weight: bold;text-decoration: underline;">HA750977</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 APP/beta Amyloid on different lysates with Rabbit anti-APP/beta Amyloid antibody (HA750977) at 1/1,000 dilution.

    Lane 1: AD Mouse brain tissue lysate
    Lane 2: Mouse brain tissue lysate (low expression)

    Lysates/proteins at 20 µg/Lane.

    Predicted band size: 87 kDa
    Observed band size: 100 kDa

    Exposure time: 3 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 (HA750977) 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;">☑ Cell treatment (CT)</span><br /><br />Immunocytochemistry analysis of HeLa cells treated with 100ng/mL Nocodazole for 17 hours labeling APP/beta Amyloid with Rabbit anti-APP/beta Amyloid antibody (<a href="/products/HA750977" style="font-weight: bold;text-decoration: underline;">HA750977</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-APP/beta Amyloid antibody (<a href="/products/HA750977" style="font-weight: bold;text-decoration: underline;">HA750977</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.

    ☑ Cell treatment (CT)

    Immunocytochemistry analysis of HeLa cells treated with 100ng/mL Nocodazole for 17 hours labeling APP/beta Amyloid with Rabbit anti-APP/beta Amyloid antibody (HA750977) 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-APP/beta Amyloid antibody (HA750977) 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 AD mouse brain tissue with Rabbit anti-APP/beta Amyloid antibody (<a href="/products/HA750977" style="font-weight: bold;text-decoration: underline;">HA750977</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/HA750977" style="font-weight: bold;text-decoration: underline;">HA750977</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 AD mouse brain tissue with Rabbit anti-APP/beta Amyloid antibody (HA750977) 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 (HA750977) 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 brain tissue with Rabbit anti-APP/beta Amyloid antibody (<a href="/products/HA750977" style="font-weight: bold;text-decoration: underline;">HA750977</a>) at 1/200 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/HA750977" style="font-weight: bold;text-decoration: underline;">HA750977</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 mouse brain tissue with Rabbit anti-APP/beta Amyloid antibody (HA750977) at 1/200 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 (HA750977) 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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