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MSR1 / CD204 Recombinant Rabbit Monoclonal Antibody [PSH29-03]

Usd: 385

Specification

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Catalog# HA724831

MSR1 / CD204 Recombinant Rabbit Monoclonal Antibody [PSH29-03]

Application

  • WB

  • IF-Cell

  • IHC-P

  • FC

  • IP

Reactivity

  • Mouse

BSA and Azide free
Conjugation

  • unconjugated

Overview

Product Name

MSR1 / CD204 Recombinant Rabbit Monoclonal Antibody [PSH29-03]

Antibody Type

Recombinant Rabbit monoclonal Antibody

Immunogen

Recombinant protein within mouse MRS1 aa 79-458.

Species Reactivity

Mouse

Validated Applications

WB, IF-Cell, IHC-P, FC, IP

Target Molecular Weight

Predicted band size: 50 kDa

Positive Control

RAW264.7 (Mouse monocytic macrophage leukemia cell) cell lysate, RAW264.7 treated with 50nM PMA and 5μg/mL Lipopolysaccharide for 24 hours cell lysate, J774A.1 (Mouse monocyte macrophage) cell lysate.

Conjugation

unconjugated

Clone Number

PSH29-03

Reactivity Data

Tested Tested Tested
Verified (internally validated)
Published Tested
Reported in literature (not internally validated)
Predicted Tested
Predicted reactive (based on sequence homology)
Not recommended Tested
Not recommended (failed internal validation)
WB IHC-P FC IF-Cell IP
human
Not Recommended
Not Recommended
Not Recommended
Not Recommended
 
mouse
Tested Tested
Tested Tested
Tested Tested
Tested Tested
Tested Tested
rat
Not Recommended
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

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

Isotype

IgG

Purification Method

Protein A affinity purified.

Application Dilution

  • WB

  • 1:5,000

  • IF-Cell

  • 1:500-1:5,000

  • IHC-P

  • 1:200-1:1,000

  • FC

  • 1:1,000

  • IP

  • 1-2μg/sample

Target

Function

Macrophage scavenger receptor 1, also known as MSR1. MSR1 has also been designated CD204 (cluster of differentiation 204). This gene encodes the class A macrophage scavenger receptors, which include three different types (1, 2, 3) generated by alternative splicing of this gene. These receptors or isoforms are trimeric integral membrane glycoproteins and have been implicated in many macrophage-associated physiological and pathological processes including atherosclerosis, Alzheimer's disease, and host defense. They were thought to be expressed macrophage-specific, but recently shown to be present on different dendritic cells classes, too. The isoforms type 1 and type 2 are functional receptors and are able to mediate the endocytosis of modified low density lipoproteins (LDLs). The isoform type 3 does not internalize modified LDL (acetyl-LDL) despite having the domain shown to mediate this function in the types 1 and 2 isoforms. It has an altered intracellular processing and is trapped within the endoplasmic reticulum, making it unable to perform endocytosis. The isoform type 3 can inhibit the function of isoforms type 1 and type 2 when co-expressed, indicating a dominant negative effect and suggesting a mechanism for regulation of scavenger receptor activity in macrophages.

Background References

1. Sheng W et al. Role of macrophage scavenger receptor MSR1 in the progression of non-alcoholic steatohepatitis. Front Immunol. 2022 Dec

2. Gudgeon J et al. The role of macrophage scavenger receptor 1 (MSR1) in inflammatory disorders and cancer. Front Immunol. 2022 Oct

Subcellular Location

Membrane.

UNIPROT

SwissProt: P30204 Mouse

Synonyms

CD204 antibody

CD204 antigen antibody

Macrophage acetylated LDL receptor I and II antibody

Macrophage scavenger receptor 1 antibody

Macrophage scavenger receptor type III antibody

Macrophage scavenger receptor types I and II antibody

Msr 1 antibody

MSR1 antibody

phSR1 antibody

phSR2 antibody

Expand

Images

  • <span style="font-weight: bold;">☑ Cell treatment (CT)</span><br /><br />Western blot analysis of MSR1 / CD204 on different lysates with Rabbit anti-MSR1 / CD204 antibody (<a href="/products/HA724831" style="font-weight: bold;text-decoration: underline;">HA724831</a>) at 1/5,000 dilution. <br /> <br />Lane 1: RAW264.7 (Mouse monocytic macrophage leukemia cell) cell lysate <br />Lane 2: RAW264.7 treated with 50nM PMA and 5μg/mL Lipopolysaccharide for 24 hours cell lysate <br /> <br />Lysates/proteins at 20 µg/Lane. <br />Exposure time: 3 minutes; ECL: K1801 <br /> <br />Blocking: 5% NFDM/TBST, 1 hour at room temperature <br />Primary antibody: <a href="/products/HA724831" style="font-weight: bold;text-decoration: underline;">HA724831</a>, 1/5,000 in primary antibody dilution buffer (<a href="/products/K1803" style="font-weight: bold;text-decoration: underline;">K1803</a>), overnight at 4 °C <br />Secondary antibody: Goat anti-Rabbit IgG-HRP (<a href="/products/HA1001" style="font-weight: bold;text-decoration: underline;">HA1001</a>), 1/50,000 in 5% NFDM/TBST, 1 hour at room temperature <br /> <br />Predicted band size: 50 kDa <br />Observed band size: 70-90 kDa

    ☑ Cell treatment (CT)

    Western blot analysis of MSR1 / CD204 on different lysates with Rabbit anti-MSR1 / CD204 antibody (HA724831) at 1/5,000 dilution.

    Lane 1: RAW264.7 (Mouse monocytic macrophage leukemia cell) cell lysate
    Lane 2: RAW264.7 treated with 50nM PMA and 5μg/mL Lipopolysaccharide for 24 hours cell lysate

    Lysates/proteins at 20 µg/Lane.
    Exposure time: 3 minutes; ECL: K1801

    Blocking: 5% NFDM/TBST, 1 hour at room temperature
    Primary antibody: HA724831, 1/5,000 in primary antibody dilution buffer (K1803), overnight at 4 °C
    Secondary antibody: Goat anti-Rabbit IgG-HRP (HA1001), 1/50,000 in 5% NFDM/TBST, 1 hour at room temperature

    Predicted band size: 50 kDa
    Observed band size: 70-90 kDa

  • <span style="font-weight: bold;">☑ Relative expression (RE)</span><br /><br />Western blot analysis of MSR1 / CD204 on different lysates with Rabbit anti-MSR1 / CD204 antibody (<a href="/products/HA724831" style="font-weight: bold;text-decoration: underline;">HA724831</a>) at 1/5,000 dilution. <br /> <br />Lane 1: J774A.1 (Mouse monocyte macrophage) cell lysate <br />Lane 2: Neuro-2a (Mouse brain neuroblastoma cell) cell lysate <br />Lane 3: NIH/3T3 (Mouse fibroblast) cell lysate <br /> <br />Lysates/proteins at 30 µg/Lane. <br />Exposure time: 3 minutes; ECL: K1802 <br /> <br />Negative control: Neuro-2a, NIH/3T3 (PMID: 35937296). <br /> <br />Blocking: 5% NFDM/TBST, 1 hour at room temperature <br />Primary antibody: <a href="/products/HA724831" style="font-weight: bold;text-decoration: underline;">HA724831</a>, 1/5,000 in primary antibody dilution buffer (<a href="/products/K1803" style="font-weight: bold;text-decoration: underline;">K1803</a>), overnight at 4 °C <br />Secondary antibody: Goat anti-Rabbit IgG-HRP (<a href="/products/HA1001" style="font-weight: bold;text-decoration: underline;">HA1001</a>), 1/50,000 in 5% NFDM/TBST, 1 hour at room temperature <br /> <br />Predicted band size: 50 kDa <br />Observed band size: 70-90 kDa

    ☑ Relative expression (RE)

    Western blot analysis of MSR1 / CD204 on different lysates with Rabbit anti-MSR1 / CD204 antibody (HA724831) at 1/5,000 dilution.

    Lane 1: J774A.1 (Mouse monocyte macrophage) cell lysate
    Lane 2: Neuro-2a (Mouse brain neuroblastoma cell) cell lysate
    Lane 3: NIH/3T3 (Mouse fibroblast) cell lysate

    Lysates/proteins at 30 µg/Lane.
    Exposure time: 3 minutes; ECL: K1802

    Negative control: Neuro-2a, NIH/3T3 (PMID: 35937296).

    Blocking: 5% NFDM/TBST, 1 hour at room temperature
    Primary antibody: HA724831, 1/5,000 in primary antibody dilution buffer (K1803), overnight at 4 °C
    Secondary antibody: Goat anti-Rabbit IgG-HRP (HA1001), 1/50,000 in 5% NFDM/TBST, 1 hour at room temperature

    Predicted band size: 50 kDa
    Observed band size: 70-90 kDa

  • <span style="font-weight: bold;">☑ Cell treatment (CT)</span><br /><br />Application: Immunocytochemistry (IF-cell) <br /> <br />Species: Mouse <br />Sample: RAW264.7 (Mouse monocytic macrophage leukemia cell) treated with or without 50 nM PMA and 5 μg/mL LPS for 24 hours <br /> <br />Fixation: 4% Paraformaldehyde, 15 minutes at room temperature. <br />Permeabilization: 0.1% Triton X-100, 15 minutes at room temperature. <br />Blocking: 1% BSA + 10% normal goat serum, 1 hour at room temperature. <br />Antibody dilution buffer: 1% BSA in PBST. <br /> <br />Primary antibody: <a href="/products/HA724831" style="font-weight: bold;text-decoration: underline;">HA724831</a>, 1/5,000, overnight at 4°C. <br />Secondary antibody: Goat Anti-Rabbit IgG (iFluor&trade; 488, <a href="/products/HA1121" style="font-weight: bold;text-decoration: underline;">HA1121</a>), 45 minutes at room temperature. <br />Counterstain: Beta tubulin (<a href="/products/HA601187" style="font-weight: bold;text-decoration: underline;">HA601187</a>, Red), 1/100, overnight at 4℃. The nuclear counterstain was DAPI (Blue).

    ☑ Cell treatment (CT)

    Application: Immunocytochemistry (IF-cell)

    Species: Mouse
    Sample: RAW264.7 (Mouse monocytic macrophage leukemia cell) treated with or without 50 nM PMA and 5 μg/mL LPS for 24 hours

    Fixation: 4% Paraformaldehyde, 15 minutes at room temperature.
    Permeabilization: 0.1% Triton X-100, 15 minutes at room temperature.
    Blocking: 1% BSA + 10% normal goat serum, 1 hour at room temperature.
    Antibody dilution buffer: 1% BSA in PBST.

    Primary antibody: HA724831, 1/5,000, overnight at 4°C.
    Secondary antibody: Goat Anti-Rabbit IgG (iFluor™ 488, HA1121), 45 minutes at room temperature.
    Counterstain: Beta tubulin (HA601187, Red), 1/100, overnight at 4℃. The nuclear counterstain was DAPI (Blue).

  • <span style="font-weight: bold;">☑ Relative expression (RE)</span><br /><br />Application: Immunocytochemistry (IF-cell)<br /><br />Species: Mouse <br />Sample: J774A.1 (Mouse monocyte macrophage) / NIH/3T3 (Mouse fibroblast)<br /><br />Fixation: 4% Paraformaldehyde, 15 minutes at room temperature.<br />Permeabilization: 0.1% Triton X-100, 15 minutes at room temperature.<br />Blocking: 1% BSA + 10% normal goat serum, 1 hour at room temperature.<br />Antibody dilution buffer: 1% BSA in PBST.<br /><br />Primary antibody: <a href="/products/HA724831" style="font-weight: bold;text-decoration: underline;">HA724831</a>, 1/500, overnight at 4℃.<br />Secondary antibody: Goat Anti-Rabbit IgG (iFluor&trade; 488, <a href="/products/HA1121" style="font-weight: bold;text-decoration: underline;">HA1121</a>), 45 minutes at room temperature.<br />Counterstain: Beta tubulin (<a href="/products/HA601187" style="font-weight: bold;text-decoration: underline;">HA601187</a>, Red), 1/100, overnight at 4℃. The nuclear counterstain was DAPI (Blue).<br /><br />Negative control: NIH/3T3 (PMID: 35937296).

    ☑ Relative expression (RE)

    Application: Immunocytochemistry (IF-cell)

    Species: Mouse
    Sample: J774A.1 (Mouse monocyte macrophage) / NIH/3T3 (Mouse fibroblast)

    Fixation: 4% Paraformaldehyde, 15 minutes at room temperature.
    Permeabilization: 0.1% Triton X-100, 15 minutes at room temperature.
    Blocking: 1% BSA + 10% normal goat serum, 1 hour at room temperature.
    Antibody dilution buffer: 1% BSA in PBST.

    Primary antibody: HA724831, 1/500, overnight at 4℃.
    Secondary antibody: Goat Anti-Rabbit IgG (iFluor™ 488, HA1121), 45 minutes at room temperature.
    Counterstain: Beta tubulin (HA601187, Red), 1/100, overnight at 4℃. The nuclear counterstain was DAPI (Blue).

    Negative control: NIH/3T3 (PMID: 35937296).

  • Application: Immunohistochemistry (IHC-P) <br /> <br />Species: Mouse <br />Tissue: Thymus <br />Sample: Paraffin-embedded section <br /> <br />Antigen retrieval: Heat-mediated, Tris-EDTA buffer (pH 9.0), 20 minutes at 95°C. <br /> <br />Wash buffer: 1× TBST <br />Endogenous peroxidase blocking: 3% H₂O₂, 10 minutes at room temperature. <br />Blocking: 1% BSA + 10% normal goat serum, 10 minutes at room temperature. <br />Primary antibody: <a href="/products/HA724831" style="font-weight: bold;text-decoration: underline;">HA724831</a>, 1/200, 1 hour at room temperature. <br />Secondary antibody: <a href="/products/HA1119" style="font-weight: bold;text-decoration: underline;">HA1119</a>, 20 minutes at room temperature.

    Application: Immunohistochemistry (IHC-P)

    Species: Mouse
    Tissue: Thymus
    Sample: Paraffin-embedded section

    Antigen retrieval: Heat-mediated, Tris-EDTA buffer (pH 9.0), 20 minutes at 95°C.

    Wash buffer: 1× TBST
    Endogenous peroxidase blocking: 3% H₂O₂, 10 minutes at room temperature.
    Blocking: 1% BSA + 10% normal goat serum, 10 minutes at room temperature.
    Primary antibody: HA724831, 1/200, 1 hour at room temperature.
    Secondary antibody: HA1119, 20 minutes at room temperature.

  • Application: Immunohistochemistry (IHC-P) <br /> <br />Species: Mouse <br />Tissue: Spleen <br />Sample: Paraffin-embedded section <br /> <br />Antigen retrieval: Heat-mediated, Tris-EDTA buffer (pH 9.0), 20 minutes at 95°C. <br /> <br />Wash buffer: 1× TBST <br />Endogenous peroxidase blocking: 3% H₂O₂, 10 minutes at room temperature. <br />Blocking: 1% BSA + 10% normal goat serum, 10 minutes at room temperature. <br />Primary antibody: <a href="/products/HA724831" style="font-weight: bold;text-decoration: underline;">HA724831</a>, 1/200, 1 hour at room temperature. <br />Secondary antibody: <a href="/products/HA1119" style="font-weight: bold;text-decoration: underline;">HA1119</a>, 20 minutes at room temperature.

    Application: Immunohistochemistry (IHC-P)

    Species: Mouse
    Tissue: Spleen
    Sample: Paraffin-embedded section

    Antigen retrieval: Heat-mediated, Tris-EDTA buffer (pH 9.0), 20 minutes at 95°C.

    Wash buffer: 1× TBST
    Endogenous peroxidase blocking: 3% H₂O₂, 10 minutes at room temperature.
    Blocking: 1% BSA + 10% normal goat serum, 10 minutes at room temperature.
    Primary antibody: HA724831, 1/200, 1 hour at room temperature.
    Secondary antibody: HA1119, 20 minutes at room temperature.

  • Application: Immunohistochemistry (IHC-P) <br /> <br />Species: Mouse <br />Tissue: Colon <br />Sample: Paraffin-embedded section <br /> <br />Antigen retrieval: Heat-mediated, Tris-EDTA buffer (pH 9.0), 20 minutes at 95°C. <br /> <br />Wash buffer: 1× TBST <br />Endogenous peroxidase blocking: 3% H₂O₂, 10 minutes at room temperature. <br />Blocking: 1% BSA + 10% normal goat serum, 10 minutes at room temperature. <br />Primary antibody: <a href="/products/HA724831" style="font-weight: bold;text-decoration: underline;">HA724831</a>, 1/200, 1 hour at room temperature. <br />Secondary antibody: <a href="/products/HA1119" style="font-weight: bold;text-decoration: underline;">HA1119</a>, 20 minutes at room temperature.

    Application: Immunohistochemistry (IHC-P)

    Species: Mouse
    Tissue: Colon
    Sample: Paraffin-embedded section

    Antigen retrieval: Heat-mediated, Tris-EDTA buffer (pH 9.0), 20 minutes at 95°C.

    Wash buffer: 1× TBST
    Endogenous peroxidase blocking: 3% H₂O₂, 10 minutes at room temperature.
    Blocking: 1% BSA + 10% normal goat serum, 10 minutes at room temperature.
    Primary antibody: HA724831, 1/200, 1 hour at room temperature.
    Secondary antibody: HA1119, 20 minutes at room temperature.

  • Application: Immunohistochemistry (IHC-P) <br /> <br />Species: Mouse <br />Tissue: RAW264.7 (Mouse monocytic macrophage leukemia cell) <br />Sample: Paraffin-embedded section <br /> <br />Antigen retrieval: Heat-mediated, Tris-EDTA buffer (pH 9.0), 20 minutes at 95°C. <br /> <br />Wash buffer: 1× TBST <br />Endogenous peroxidase blocking: 3% H₂O₂, 10 minutes at room temperature. <br />Blocking: 1% BSA + 10% normal goat serum, 10 minutes at room temperature. <br />Primary antibody: <a href="/products/HA724831" style="font-weight: bold;text-decoration: underline;">HA724831</a>, 1/1,000, 1 hour at room temperature. <br />Secondary antibody: <a href="/products/HA1119" style="font-weight: bold;text-decoration: underline;">HA1119</a>, 20 minutes at room temperature.

    Application: Immunohistochemistry (IHC-P)

    Species: Mouse
    Tissue: RAW264.7 (Mouse monocytic macrophage leukemia cell)
    Sample: Paraffin-embedded section

    Antigen retrieval: Heat-mediated, Tris-EDTA buffer (pH 9.0), 20 minutes at 95°C.

    Wash buffer: 1× TBST
    Endogenous peroxidase blocking: 3% H₂O₂, 10 minutes at room temperature.
    Blocking: 1% BSA + 10% normal goat serum, 10 minutes at room temperature.
    Primary antibody: HA724831, 1/1,000, 1 hour at room temperature.
    Secondary antibody: HA1119, 20 minutes at room temperature.

  • Application: Immunohistochemistry (IHC-P) <br /> <br />Species: Mouse <br />Tissue: J774A.1 (Mouse monocyte macrophage) <br />Sample: Paraffin-embedded section <br /> <br />Antigen retrieval: Heat-mediated, Tris-EDTA buffer (pH 9.0), 20 minutes at 95°C. <br /> <br />Wash buffer: 1× TBST <br />Endogenous peroxidase blocking: 3% H₂O₂, 10 minutes at room temperature. <br />Blocking: 1% BSA + 10% normal goat serum, 10 minutes at room temperature. <br />Primary antibody: <a href="/products/HA724831" style="font-weight: bold;text-decoration: underline;">HA724831</a>, 1/1,000, 1 hour at room temperature. <br />Secondary antibody: <a href="/products/HA1119" style="font-weight: bold;text-decoration: underline;">HA1119</a>, 20 minutes at room temperature.

    Application: Immunohistochemistry (IHC-P)

    Species: Mouse
    Tissue: J774A.1 (Mouse monocyte macrophage)
    Sample: Paraffin-embedded section

    Antigen retrieval: Heat-mediated, Tris-EDTA buffer (pH 9.0), 20 minutes at 95°C.

    Wash buffer: 1× TBST
    Endogenous peroxidase blocking: 3% H₂O₂, 10 minutes at room temperature.
    Blocking: 1% BSA + 10% normal goat serum, 10 minutes at room temperature.
    Primary antibody: HA724831, 1/1,000, 1 hour at room temperature.
    Secondary antibody: HA1119, 20 minutes at room temperature.

  • <span style="font-weight: bold;">☑ Relative expression (RE)</span><br /><br />Application: Immunohistochemistry (IHC-P) <br /> <br />Species: Mouse <br />Tissue: Neuro-2a (Mouse brain neuroblastoma cell) <br />Sample: Paraffin-embedded section <br /> <br />Negative control: Neuro-2a (PMID: 35937296). <br /> <br />Antigen retrieval: Heat-mediated, Tris-EDTA buffer (pH 9.0), 20 minutes at 95°C. <br /> <br />Wash buffer: 1× TBST <br />Endogenous peroxidase blocking: 3% H₂O₂, 10 minutes at room temperature. <br />Blocking: 1% BSA + 10% normal goat serum, 10 minutes at room temperature. <br />Primary antibody: <a href="/products/HA724831" style="font-weight: bold;text-decoration: underline;">HA724831</a>, 1/1,000, 1 hour at room temperature. <br />Secondary antibody: <a href="/products/HA1119" style="font-weight: bold;text-decoration: underline;">HA1119</a>, 20 minutes at room temperature.

    ☑ Relative expression (RE)

    Application: Immunohistochemistry (IHC-P)

    Species: Mouse
    Tissue: Neuro-2a (Mouse brain neuroblastoma cell)
    Sample: Paraffin-embedded section

    Negative control: Neuro-2a (PMID: 35937296).

    Antigen retrieval: Heat-mediated, Tris-EDTA buffer (pH 9.0), 20 minutes at 95°C.

    Wash buffer: 1× TBST
    Endogenous peroxidase blocking: 3% H₂O₂, 10 minutes at room temperature.
    Blocking: 1% BSA + 10% normal goat serum, 10 minutes at room temperature.
    Primary antibody: HA724831, 1/1,000, 1 hour at room temperature.
    Secondary antibody: HA1119, 20 minutes at room temperature.

  • <span style="font-weight: bold;">☑ Relative expression (RE)</span><br /><br />Application: Flow Cytometry <br /> <br />Species: Mouse <br />Sample: NIH/3T3 (Mouse fibroblasts) (Left, negative) / J774A.1 (Mouse monocyte macrophage) <br /> <br />Blocking: 1% BSA + 10% normal goat serum, 15 minutes at room temperature. <br />Antibody dilution buffer: 1x PBS. <br /> <br />Primary antibody: <a href="/products/HA724831" style="font-weight: bold;text-decoration: underline;">HA724831</a>(1/1,000, Red) compared with Rabbit IgG Isotype Control (<a href="/products/HA722127" style="font-weight: bold;text-decoration: underline;">HA722127</a>, Green), 15 minutes at room temperature. <br />Secondary antibody: Goat Anti-Rabbit IgG (iFluor&trade; 488, <a href="/products/HA1121" style="font-weight: bold;text-decoration: underline;">HA1121</a>), 15 minutes at room temperature. <br /> <br />Negative control: NIH/3T3 (PMID: 35937296).

    ☑ Relative expression (RE)

    Application: Flow Cytometry

    Species: Mouse
    Sample: NIH/3T3 (Mouse fibroblasts) (Left, negative) / J774A.1 (Mouse monocyte macrophage)

    Blocking: 1% BSA + 10% normal goat serum, 15 minutes at room temperature.
    Antibody dilution buffer: 1x PBS.

    Primary antibody: HA724831(1/1,000, Red) compared with Rabbit IgG Isotype Control (HA722127, Green), 15 minutes at room temperature.
    Secondary antibody: Goat Anti-Rabbit IgG (iFluor™ 488, HA1121), 15 minutes at room temperature.

    Negative control: NIH/3T3 (PMID: 35937296).

  • <span style="font-weight: bold;">☑ Cell treatment (CT)</span><br /><br />Immunoprecipitation (IP)<br /><br />MSR1 / CD204 was immunoprecipitated in 0.2 mg RAW264.7 (Mouse monocytic macrophage leukemia cell) treated with 50nM PMA and 5μg/mL LPS for 24 hours cell lysate with <a href="/products/HA724831" style="font-weight: bold;text-decoration: underline;">HA724831</a> at 2 µg/10 µl beads. Western blot was performed from the immunoprecipitate using <a href="/products/HA724831" style="font-weight: bold;text-decoration: underline;">HA724831</a> at 1/5,000 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: RAW264.7 treated with 50nM PMA and 5μg/mL LPS for 24 hours cell lysate (input)<br />Lane 2: <a href="/products/HA724831" style="font-weight: bold;text-decoration: underline;">HA724831</a> IP in RAW264.7 treated with 50nM PMA and 5μg/mL LPS for 24 hours cell lysate<br />Lane 3: Rabbit IgG instead of <a href="/products/HA724831" style="font-weight: bold;text-decoration: underline;">HA724831</a> in RAW264.7 treated with 50nM PMA and 5μg/mL LPS for 24 hours cell lysate<br /><br />Exposure time: 2 seconds<br />Blocking: 5% NFDM/TBST, 1 hour at room temperature<br /><br />Primary dilution: <a href="/products/HA724831" style="font-weight: bold;text-decoration: underline;">HA724831</a>, 1/5,000 in primary antibody dilution buffer (<a href="/products/K1803" style="font-weight: bold;text-decoration: underline;">K1803</a>), 2 hours at room temperature<br /><br />Predicted band size: 50 kDa<br />Observed band size: 70-90 kDa

    ☑ Cell treatment (CT)

    Immunoprecipitation (IP)

    MSR1 / CD204 was immunoprecipitated in 0.2 mg RAW264.7 (Mouse monocytic macrophage leukemia cell) treated with 50nM PMA and 5μg/mL LPS for 24 hours cell lysate with HA724831 at 2 µg/10 µl beads. Western blot was performed from the immunoprecipitate using HA724831 at 1/5,000 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: RAW264.7 treated with 50nM PMA and 5μg/mL LPS for 24 hours cell lysate (input)
    Lane 2: HA724831 IP in RAW264.7 treated with 50nM PMA and 5μg/mL LPS for 24 hours cell lysate
    Lane 3: Rabbit IgG instead of HA724831 in RAW264.7 treated with 50nM PMA and 5μg/mL LPS for 24 hours cell lysate

    Exposure time: 2 seconds
    Blocking: 5% NFDM/TBST, 1 hour at room temperature

    Primary dilution: HA724831, 1/5,000 in primary antibody dilution buffer (K1803), 2 hours at room temperature

    Predicted band size: 50 kDa
    Observed band size: 70-90 kDa

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