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F4/80 Recombinant Rabbit Monoclonal Antibody [PSH0-85]

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Specification

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Catalog# HA721520

F4/80 Recombinant Rabbit Monoclonal Antibody [PSH0-85]

Application

  • WB

  • IHC-P

  • IF-Tissue

  • IHC-Fr

  • mIHC

Reactivity

  • Mouse

  • Rat

BSA and Azide free
Conjugation

  • unconjugated

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

Overview

Product Name

F4/80 Recombinant Rabbit Monoclonal Antibody [PSH0-85]

Antibody Type

Recombinant Rabbit monoclonal Antibody

Immunogen

Recombinant protein within mouse F4/80 aa 1-650 / 931.

Species Reactivity

Mouse, Rat

Validated Applications

WB, IHC-P, IF-Tissue, IHC-Fr, mIHC

Molecular Weight

Predicted band size: 102 kDa

Positive Control

Mouse kidney tissue lysate, RAW264.7 cell lysate, Rat kidney tissue lysate, Rat spleen tissue lysate, mouse liver tissue, mouse spleen tissue, rat liver tissue, rat spleen tissue, mouse lung tissue, rat lung tissue.

Conjugation

unconjugated

Clone Number

PSH0-85

RRID

AB_3072636

Reactivity Data

WB IHC-P IF-Tissue IHC-Fr mIHC
Mouse
Tested Tested
Tested Tested
Tested Tested
Tested Tested
Tested
Rat
Tested Tested
Tested Tested
Tested
Tested Tested

Product Features

Form

Liquid

Concentration

Storage Instructions

Shipped at 4℃. Store at +4℃ short term (1-2 weeks). It is recommended to aliquot into single-use upon delivery. Store at -20℃ long term.

Storage Buffer

PBS (pH7.4), 0.1% BSA, 40% Glycerol. Preservative: 0.05% Sodium Azide.

Isotype

IgG

Purification Method

Protein A affinity purified.

Application Dilution

  • WB

  • 1:1,000

  • IHC-P

  • 1:200-1:1,000

  • IF-Tissue

  • 1:500

  • IHC-Fr

  • 1:1,000

  • mIHC

  • 1:500

Target

Function

EGF-like module-containing mucin-like hormone receptor-like 1 also known as F4/80 is a protein encoded by the ADGRE1 gene. EMR1 is a member of the adhesion GPCR family. Adhesion GPCRs are characterized by an extended extracellular region often possessing N-terminal protein modules that is linked to a TM7 region via a domain known as the GPCR-Autoproteolysis INducing (GAIN) domain. EMR1 expression in human is restricted to eosinophils and is a specific marker for these cells. The murine homolog of EMR1, F4/80, is a well-known and widely used marker of murine macrophage populations. The N-terminal fragment (NTF) of EMR1 contains 4-6 Epidermal Growth Factor-like (EGF-like) domains in human and 4-7 EGF-like domains in the mouse. Utilizing F4/80 knockout mice, Lin et al. showed that F4/80 is not necessary for the development of tissue macrophages but is required for the induction of efferent CD8+ regulatory T cells needed for peripheral tolerance.

Background References

1. Deng R et al. Periosteal CD68+ F4/80+ Macrophages Are Mechanosensitive for Cortical Bone Formation by Secretion and Activation of TGF-β1. Adv Sci (Weinh). 2022 Jan

2. Shin AE et al. F4/80+Ly6Chigh Macrophages Lead to Cell Plasticity and Cancer Initiation in Colitis. Gastroenterology. 2023 Apr

Subcellular Location

Cell membrane.

UNIPROT

SwissProt: Q61549 Mouse

SwissProt: Q5Y4N8 Rat

Synonyms

ADGRE1 antibody

Adhesion G protein coupled receptor E1 antibody

Adhesion G protein-coupled receptor E1 antibody

AGRE1_HUMAN antibody

Cell surface glycoprotein EMR1 antibody

Cell surface glycoprotein F4/80 antibody

DD7A5 7 antibody

Egf like module containing mucin like hormone receptor like 1 antibody

Egf like module containing mucin like hormone receptor like sequence 1 antibody

EGF like module receptor 1 antibody

Expand

Images

  • Western blot analysis of F4/80 on different lysates with Rabbit anti-F4/80 antibody (<a href="/products/HA721520" style="font-weight: bold;text-decoration: underline;">HA721520</a>) at 1/1,000 dilution.<br /><br />Lane 1: Mouse kidney tissue lysate (no heat) (40 µg/Lane) <br />Lane 2: Mouse kidney tissue lysate (40 µg/Lane) <br />Lane 3: RAW264.7 cell lysate (no heat) (20 µg/Lane)<br />Lane 4: Rat kidney tissue lysate (no heat) (40 µg/Lane) <br /><br />Notice: no heat means the lysate is not boiled.<br /><br />Predicted band size: 102 kDa<br />Observed band size: 160 kDa<br /><br />Exposure time: 59 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/HA721520" style="font-weight: bold;text-decoration: underline;">HA721520</a>) at 1/1,000 dilution was used in primary antibody dilution (K1803) 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 F4/80 on different lysates with Rabbit anti-F4/80 antibody (HA721520) at 1/1,000 dilution.

    Lane 1: Mouse kidney tissue lysate (no heat) (40 µg/Lane)
    Lane 2: Mouse kidney tissue lysate (40 µg/Lane)
    Lane 3: RAW264.7 cell lysate (no heat) (20 µg/Lane)
    Lane 4: Rat kidney tissue lysate (no heat) (40 µg/Lane)

    Notice: no heat means the lysate is not boiled.

    Predicted band size: 102 kDa
    Observed band size: 160 kDa

    Exposure time: 59 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 (HA721520) at 1/1,000 dilution was used in primary antibody dilution (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.

  • <span style="font-weight: bold;">☑ Relative expression (RE)</span><br /><br />Western blot analysis of F4/80 on different lysates with Rabbit anti-F4/80 antibody (<a href="/products/HA721520" style="font-weight: bold;text-decoration: underline;">HA721520</a>) at 1/1,000 dilution.<br /><br />Lane 1: RAW264.7 cell lysate (no heat) (20 µg/Lane)<br />Lane 2: L929 cell lysate (no heat) (negative) (20 µg/Lane)<br />Lane 3: Rat spleen tissue lysate (70℃ heat) (40 µg/Lane)<br /><br />Notice: no heat means the lysate is not boiled.<br /><br />Predicted band size: 102 kDa<br />Observed band size: 160 kDa<br /><br />Exposure time: 2 minutes;<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/HA721520" style="font-weight: bold;text-decoration: underline;">HA721520</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 1:100,000 dilution was used for 1 hour at room temperature.

    ☑ Relative expression (RE)

    Western blot analysis of F4/80 on different lysates with Rabbit anti-F4/80 antibody (HA721520) at 1/1,000 dilution.

    Lane 1: RAW264.7 cell lysate (no heat) (20 µg/Lane)
    Lane 2: L929 cell lysate (no heat) (negative) (20 µg/Lane)
    Lane 3: Rat spleen tissue lysate (70℃ heat) (40 µg/Lane)

    Notice: no heat means the lysate is not boiled.

    Predicted band size: 102 kDa
    Observed band size: 160 kDa

    Exposure time: 2 minutes;

    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 (HA721520) at 1/1,000 dilution was used in 5% NFDM/TBST at 4℃ overnight. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1:100,000 dilution was used for 1 hour at room temperature.

  • Application: IHC-Fr<br /><br />Species: Mouse<br /><br />Site: Spleen<br /><br />Sample: Frozen section<br /><br />Antibody concentration: 1/1,000 (F4/80, <a href="/products/HA721520" style="font-weight: bold;text-decoration: underline;">HA721520</a>, rabbit, red); 1/500 (Iba1, <a href="/products/HA601376" style="font-weight: bold;text-decoration: underline;">HA601376</a>, Chicken, green)<br /> <br />Antigen retrieval: Not required

    Application: IHC-Fr

    Species: Mouse

    Site: Spleen

    Sample: Frozen section

    Antibody concentration: 1/1,000 (F4/80, HA721520, rabbit, red); 1/500 (Iba1, HA601376, Chicken, green)

    Antigen retrieval: Not required

  • Application: IHC-Fr<br /><br />Species: Rat<br /><br />Site: Spleen<br /><br />Sample: Frozen section<br /><br />Antibody concentration: 1/1,000 (F4/80, <a href="/products/HA721520" style="font-weight: bold;text-decoration: underline;">HA721520</a>, rabbit, red); 1/500 (Iba1, <a href="/products/HA601376" style="font-weight: bold;text-decoration: underline;">HA601376</a>, Chicken, green)<br /> <br />Antigen retrieval: Not required

    Application: IHC-Fr

    Species: Rat

    Site: Spleen

    Sample: Frozen section

    Antibody concentration: 1/1,000 (F4/80, HA721520, rabbit, red); 1/500 (Iba1, HA601376, Chicken, green)

    Antigen retrieval: Not required

  • Immunohistochemical analysis of paraffin-embedded mouse liver tissue with Rabbit anti-F4/80 antibody (<a href="/products/HA721520" style="font-weight: bold;text-decoration: underline;">HA721520</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/HA721520" style="font-weight: bold;text-decoration: underline;">HA721520</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 mouse liver tissue with Rabbit anti-F4/80 antibody (HA721520) 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 (HA721520) 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 spleen tissue with Rabbit anti-F4/80 antibody (<a href="/products/HA721520" style="font-weight: bold;text-decoration: underline;">HA721520</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/HA721520" style="font-weight: bold;text-decoration: underline;">HA721520</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 mouse spleen tissue with Rabbit anti-F4/80 antibody (HA721520) 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 (HA721520) 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 rat liver tissue with Rabbit anti-F4/80 antibody (<a href="/products/HA721520" style="font-weight: bold;text-decoration: underline;">HA721520</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/HA721520" style="font-weight: bold;text-decoration: underline;">HA721520</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 rat liver tissue with Rabbit anti-F4/80 antibody (HA721520) 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 (HA721520) 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 rat spleen tissue with Rabbit anti-F4/80 antibody (<a href="/products/HA721520" style="font-weight: bold;text-decoration: underline;">HA721520</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/HA721520" style="font-weight: bold;text-decoration: underline;">HA721520</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 rat spleen tissue with Rabbit anti-F4/80 antibody (HA721520) 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 (HA721520) 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 lung tissue with Rabbit anti-F4/80 antibody (<a href="/products/HA721520" style="font-weight: bold;text-decoration: underline;">HA721520</a>) at 1/500 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/HA721520" style="font-weight: bold;text-decoration: underline;">HA721520</a>) at 1/500 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 lung tissue with Rabbit anti-F4/80 antibody (HA721520) at 1/500 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 (HA721520) at 1/500 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 lung tissue with Rabbit anti-F4/80 antibody (<a href="/products/HA721520" style="font-weight: bold;text-decoration: underline;">HA721520</a>) at 1/2,000 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/HA721520" style="font-weight: bold;text-decoration: underline;">HA721520</a>) at 1/2,000 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 lung tissue with Rabbit anti-F4/80 antibody (HA721520) at 1/2,000 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 (HA721520) at 1/2,000 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.

  • Application: IF-tissue<br /><br />Species: Mouse<br /><br />Site: Liver<br /><br />Sample: Paraffin-embedded section<br /><br />Antibody concentration: 1/500

    Application: IF-tissue

    Species: Mouse

    Site: Liver

    Sample: Paraffin-embedded section

    Antibody concentration: 1/500

  • Application: IF-tissue<br /><br />Species: Rat<br /><br />Site: Liver<br /><br />Sample: Paraffin-embedded section<br /><br />Antibody concentration: 1/500

    Application: IF-tissue

    Species: Rat

    Site: Liver

    Sample: Paraffin-embedded section

    Antibody concentration: 1/500

  • Application: IF-tissue<br /><br />Species: Mouse<br /><br />Site: spleen<br /><br />Sample: Paraffin-embedded section<br /><br />Antibody concentration: 1/500

    Application: IF-tissue

    Species: Mouse

    Site: spleen

    Sample: Paraffin-embedded section

    Antibody concentration: 1/500

  • Application: IF-tissue<br /><br />Species: Mouse<br /><br />Site: small intestine<br /><br />Sample: Paraffin-embedded section<br /><br />Antibody concentration: 1/500

    Application: IF-tissue

    Species: Mouse

    Site: small intestine

    Sample: Paraffin-embedded section

    Antibody concentration: 1/500

  • mIHC analysis of mouse spleen tissue (Formalin/PFA-fixed paraffin-embedded sections) with Rabbit anti-F4/80 antibody (<a href="/products/HA721520" style="font-weight: bold;text-decoration: underline;">HA721520</a>) at 1/500 dilution. The immunostaining was performed with the IRISKitCmTSA Kit (<a href="/products/900808" style="font-weight: bold;text-decoration: underline;">900808</a>). Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 30 mins at 95℃. DAPI (blue) was used as a nuclear counter stain. Image acquisition was performed with Olympus VS200 Slide Scanner.

    mIHC analysis of mouse spleen tissue (Formalin/PFA-fixed paraffin-embedded sections) with Rabbit anti-F4/80 antibody (HA721520) at 1/500 dilution. The immunostaining was performed with the IRISKitCmTSA Kit (900808). Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 30 mins at 95℃. DAPI (blue) was used as a nuclear counter stain. Image acquisition was performed with Olympus VS200 Slide Scanner.

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Citation