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AFP Recombinant Rabbit Monoclonal Antibody [PSH11-75]

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Specification

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Catalog# HA723371

AFP Recombinant Rabbit Monoclonal Antibody [PSH11-75]

Application

  • WB

  • IF-Cell

  • IHC-P

  • IF-Tissue

Reactivity

  • Human

  • Mouse

  • Rat

BSA and Azide free
Conjugation

  • unconjugated

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

Overview

Product Name

AFP Recombinant Rabbit Monoclonal Antibody [PSH11-75]

Antibody Type

Recombinant Rabbit monoclonal Antibody

Immunogen

Recombinant protein within human AFP aa 1-609.

Species Reactivity

Human, Mouse, Rat

Validated Applications

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

Molecular Weight

Predicted band size: 69 kDa

Positive Control

HepG2 cell lysate, Mouse placenta tissue lysate, human placenta tissue, human poorly differentiated hepatocellular carcinoma tissue, mouse embryo liver tissue, rat embryo liver tissue, HepG2.

Conjugation

unconjugated

Clone Number

PSH11-75

Reactivity Data

WB IF-Cell IHC-P IF-Tissue
Human
Tested Tested
Tested Tested
Tested Tested
Mouse
Tested Tested
Tested Tested
Rat
Tested Tested

Product Features

Form

Liquid

Concentration

Storage Instructions

Shipped at 4℃. Store at +4℃ short term (1-2 weeks). Store at -20℃ long term.

Storage Buffer

1*PBS (pH7.4), 0.1% BSA, 40% Glycerol, 0.2% Proclean 950.

Isotype

IgG

Purification Method

Protein A affinity purified.

Application Dilution

  • WB

  • 1:2,000

  • IF-Cell

  • 1:250

  • IHC-P

  • 1:20,000

  • IF-Tissue

  • 1:1,000

Target

Function

Alpha-fetoprotein (AFP, α-fetoprotein; also sometimes called alpha-1-fetoprotein, alpha-fetoglobulin, or alpha fetal protein) is a protein that in humans is encoded by the AFP gene. The AFP gene is located on the q arm of chromosome 4 (4q13.3). Maternal AFP serum level is used to screen for Down syndrome, neural tube defects, and other chromosomal abnormalities. AFP is a major plasma protein produced by the yolk sac and the fetal liver during fetal development. It is thought to be the fetal analog of serum albumin. AFP binds to copper, nickel, fatty acids and bilirubin and is found in monomeric, dimeric and trimeric forms.

Background References

1. Yeo YH et al. Alpha-fetoprotein: Past, present, and future. Hepatol Commun. 2024 Apr

2. Hu X et al. The Landscape Of Alpha Fetoprotein In Hepatocellular Carcinoma: Where Are We? Int J Biol Sci. 2022 Jan

Subcellular Location

Secreted.

UNIPROT

SwissProt: P02771 Human

SwissProt: P02772 Mouse

SwissProt: P02773 Rat

Synonyms

Afp antibody

AFPD antibody

Alpha fetoglobulin antibody

Alpha fetoprotein antibody

Alpha fetoprotein precursor antibody

Alpha-1-fetoprotein antibody

Alpha-fetoglobulin antibody

Alpha-fetoprotein antibody

alpha-fetoprotein, Hereditary persistence of, included antibody

FETA antibody

Expand

Images

  • Western blot analysis of AFP on different lysates with Rabbit anti-AFP antibody (<a href="/products/HA723371" style="font-weight: bold;text-decoration: underline;">HA723371</a>) at 1/2,000 dilution.<br /><br />Lane 1: HepG2 cell lysate (20 µg/Lane)<br />Lane 2: Mouse placenta tissue lysate (40 µg/Lane)<br /><br />Predicted band size: 69 kDa<br />Observed band size: 69 kDa<br /><br />Exposure time: Lane 1: 25 seconds; Lane 2: 8 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/HA723371" style="font-weight: bold;text-decoration: underline;">HA723371</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 AFP on different lysates with Rabbit anti-AFP antibody (HA723371) at 1/2,000 dilution.

    Lane 1: HepG2 cell lysate (20 µg/Lane)
    Lane 2: Mouse placenta tissue lysate (40 µg/Lane)

    Predicted band size: 69 kDa
    Observed band size: 69 kDa

    Exposure time: Lane 1: 25 seconds; Lane 2: 8 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 (HA723371) 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.

  • Immunohistochemical analysis of paraffin-embedded human placenta tissue with Rabbit anti-AFP antibody (<a href="/products/HA723371" style="font-weight: bold;text-decoration: underline;">HA723371</a>) at 1/20,000 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/HA723371" style="font-weight: bold;text-decoration: underline;">HA723371</a>) at 1/20,000 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 placenta tissue with Rabbit anti-AFP antibody (HA723371) at 1/20,000 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 (HA723371) at 1/20,000 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 poorly differentiated hepatocellular carcinoma tissue with Rabbit anti-AFP antibody (<a href="/products/HA723371" style="font-weight: bold;text-decoration: underline;">HA723371</a>) at 1/20,000 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/HA723371" style="font-weight: bold;text-decoration: underline;">HA723371</a>) at 1/20,000 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 poorly differentiated hepatocellular carcinoma tissue with Rabbit anti-AFP antibody (HA723371) at 1/20,000 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 (HA723371) at 1/20,000 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.

  • <span style="font-weight: bold;">☑ Relative expression (RE)</span><br /><br />Immunohistochemical analysis of paraffin-embedded human well-differentiated hepatocellular carcinoma tissue (negative) with Rabbit anti-AFP antibody (<a href="/products/HA723371" style="font-weight: bold;text-decoration: underline;">HA723371</a>) at 1/20,000 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/HA723371" style="font-weight: bold;text-decoration: underline;">HA723371</a>) at 1/20,000 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.

    ☑ Relative expression (RE)

    Immunohistochemical analysis of paraffin-embedded human well-differentiated hepatocellular carcinoma tissue (negative) with Rabbit anti-AFP antibody (HA723371) at 1/20,000 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 (HA723371) at 1/20,000 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 embryo liver tissue with Rabbit anti-AFP antibody (<a href="/products/HA723371" style="font-weight: bold;text-decoration: underline;">HA723371</a>) at 1/20,000 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/HA723371" style="font-weight: bold;text-decoration: underline;">HA723371</a>) at 1/20,000 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 embryo liver tissue with Rabbit anti-AFP antibody (HA723371) at 1/20,000 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 (HA723371) at 1/20,000 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 embryo liver tissue with Rabbit anti-AFP antibody (<a href="/products/HA723371" style="font-weight: bold;text-decoration: underline;">HA723371</a>) at 1/20,000 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/HA723371" style="font-weight: bold;text-decoration: underline;">HA723371</a>) at 1/20,000 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 embryo liver tissue with Rabbit anti-AFP antibody (HA723371) at 1/20,000 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 (HA723371) at 1/20,000 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.

  • Immunocytochemistry analysis of HepG2 cells labeling AFP with Rabbit anti-AFP antibody (<a href="/products/HA723371" style="font-weight: bold;text-decoration: underline;">HA723371</a>) at 1/250 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-AFP antibody (<a href="/products/HA723371" style="font-weight: bold;text-decoration: underline;">HA723371</a>) at 1/250 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/HA601187" style="font-weight: bold;text-decoration: underline;">HA601187</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 HepG2 cells labeling AFP with Rabbit anti-AFP antibody (HA723371) at 1/250 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-AFP antibody (HA723371) at 1/250 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 (HA601187, 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.

  • Application: IF-Tissue<br /><br />Species: Human<br /><br />Site: poorly differentiated hepatocellular carcinoma<br /><br />Sample: Paraffin-embedded section<br /><br />Antibody concentration: 1/1,000

    Application: IF-Tissue

    Species: Human

    Site: poorly differentiated hepatocellular carcinoma

    Sample: Paraffin-embedded section

    Antibody concentration: 1/1,000

  • <span style="font-weight: bold;">☑ Relative expression (RE)</span><br /><br />Application: IF-Tissue<br /><br />Species: Human<br /><br />Site: liver (negative)<br /><br />Sample: Paraffin-embedded section<br /><br />Antibody concentration: 1/1,000

    ☑ Relative expression (RE)

    Application: IF-Tissue

    Species: Human

    Site: liver (negative)

    Sample: Paraffin-embedded section

    Antibody concentration: 1/1,000

Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"

Citation