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Human IgM Recombinant Rabbit Monoclonal Antibody [JF84-09]

Usd: 385

Specification

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Catalog# ET1702-81

Human IgM Recombinant Rabbit Monoclonal Antibody [JF84-09]

Application

  • WB

  • IHC-P

  • IP

  • FC

  • IF-Cell

Reactivity

  • Human

Conjugation

  • unconjugated

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

Overview

Product Name

Human IgM Recombinant Rabbit Monoclonal Antibody [JF84-09]

Antibody Type

Recombinant Rabbit monoclonal Antibody

Immunogen

Recombinant protein within Human IgM from Human serum (Sigma#I8260).

Species Reactivity

Human

Validated Applications

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

Molecular Weight

Predicted band size: 50 kDa

Positive Control

Daudi cell lysate, human IgM protein, human tonsil tissue, human spleen tissue, Daudi.

Conjugation

unconjugated

Clone Number

JF84-09

RRID

AB_3070341

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*TBS (pH7.4), 0.05% BSA, 40% Glycerol. Preservative: 0.05% Sodium Azide.

Isotype

IgG

Purification Method

Protein A affinity purified.

Application Dilution

  • WB

  • 1:1,000-1:5,000

  • IHC-P

  • 1:50-1:200

  • FC

  • 1:1,000

  • IF-Cell

  • 1:100

Target

Function

Immunoglobulin M (IgM) is one of several isotypes of antibody (also known as immunoglobulin) that are produced by vertebrates. IgM is the largest antibody, and it is the first antibody to appear in the response to initial exposure to an antigen. In humans and other mammals that have been studied, Plasmablasts residing in the spleen are the main source for specific IgM production. IgM interacts with several other physiological molecules: IgM can bind complement component C1 and activate the classical pathway, leading to opsonization of antigens and cytolysis; IgM binds to the polyimmunoglobulin receptor (pIgR) in a process that brings IgM to mucosal surfaces, such as the gut lumen and into breast milk. This binding depends on J chain; Two other Fc receptors that bind IgM—Fcα/µ-R and Fcµ-R -- have been detected. Fcα/µ-R, like pIgR, binds polymeric IgM and IgA. Fcα/µ-R can mediate endocytosis, and its expression in the gut suggests a role in mucosal immunity. Fcµ-R (formerly known as Toso/Faim3) binds IgM exclusively and can mediate cellular uptake of IgM-conjugated antigen.

Background References

1. Klasener K., et al. 2014. B cell activation involves nanoscale receptor reorganizations and inside-out signaling by Syk. Elife 3:e02069-e02069.

2. Huang J., et al. 2014. Rearrangement and expression of the immunoglobulin mu-chain gene in human myeloid cells. Cell. Mol. Immunol. 11:94-104.

Subcellular Location

Secreted, Cell membrane.

UNIPROT

SwissProt: P01871 Human

Synonyms

Immunoglobin heavy chain constant region mu edit item name - Immunoglobin heavy chain mu constant region antibody

Immunoglobin heavy chain constant region mu antibody

AGM1 antibody

Constant region of heavy chain of IgM antibody

DKFZp686I15196 antibody

DKFZp686I15212 antibody

FLJ00385 antibody

Ig mu chain C region antibody

IGHM antibody

IgM heavy chain constant region antibody

Expand

Images

  • Western blot analysis of Human IgM on Daudi cell lysates with Rabbit anti-Human IgM antibody (<a href="/products/ET1702-81" style="font-weight: bold;text-decoration: underline;">ET1702-81</a>) at 1/1,000 dilution.<br /><br />Lysates/proteins at 20 µg/Lane.<br /><br />Predicted band size: 50 kDa<br />Observed band size: 80 kDa<br /><br />Exposure time: 30 seconds;<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/ET1702-81" style="font-weight: bold;text-decoration: underline;">ET1702-81</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.

    Western blot analysis of Human IgM on Daudi cell lysates with Rabbit anti-Human IgM antibody (ET1702-81) at 1/1,000 dilution.

    Lysates/proteins at 20 µg/Lane.

    Predicted band size: 50 kDa
    Observed band size: 80 kDa

    Exposure time: 30 seconds;

    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 (ET1702-81) 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 />Western blot analysis of Human IgM on different lysates with Rabbit anti-Human IgM antibody (<a href="/products/ET1702-81" style="font-weight: bold;text-decoration: underline;">ET1702-81</a>) at 1/1,000 dilution.<br /><br />Lane 1: Daudi cell lysate<br />Lane 2: Daudi treated with deglycosylation cell lysate<br /><br />Lysates/proteins at 20 µg/Lane.<br /><br />Predicted band size: 50 kDa<br />Observed band size: 80 kDa<br /><br />Exposure time: 30 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/ET1702-81" style="font-weight: bold;text-decoration: underline;">ET1702-81</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 Human IgM on different lysates with Rabbit anti-Human IgM antibody (ET1702-81) at 1/1,000 dilution.

    Lane 1: Daudi cell lysate
    Lane 2: Daudi treated with deglycosylation cell lysate

    Lysates/proteins at 20 µg/Lane.

    Predicted band size: 50 kDa
    Observed band size: 80 kDa

    Exposure time: 30 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 (ET1702-81) 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.

  • Immunohistochemical analysis of paraffin-embedded human tonsil tissue using anti-Human IgM antibody. 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 30 minutes at room temperature, washed with ddH<sub>2</sub>O and PBS, and then probed with the primary antibody (<a href="/products/ET1702-81" style="font-weight: bold;text-decoration: underline;">ET1702-81</a>, 1/50) for 30 minutes 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 tonsil tissue using anti-Human IgM antibody. 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 30 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (ET1702-81, 1/50) for 30 minutes 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 spleen tissue using anti-Human IgM antibody. 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 30 minutes at room temperature, washed with ddH<sub>2</sub>O and PBS, and then probed with the primary antibody (<a href="/products/ET1702-81" style="font-weight: bold;text-decoration: underline;">ET1702-81</a>, 1/50) for 30 minutes 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 spleen tissue using anti-Human IgM antibody. 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 30 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (ET1702-81, 1/50) for 30 minutes 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.

  • Flow cytometric analysis of Daudi cells labeling Human IgM.<br /><br />Cells were fixed and permeabilized. Then stained with the primary antibody (<a href="/products/ET1702-81" style="font-weight: bold;text-decoration: underline;">ET1702-81</a>, 1μg/mL) (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 Daudi cells labeling Human IgM.

    Cells were fixed and permeabilized. Then stained with the primary antibody (ET1702-81, 1μg/mL) (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).

  • Immunocytochemistry analysis of Daudi cells labeling Human IgM with Rabbit anti-Human IgM antibody (<a href="/products/ET1702-81" style="font-weight: bold;text-decoration: underline;">ET1702-81</a>) at 1/100 dilution.<br /><br />Cells were fixed in 80% precooled methanol 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-Human IgM antibody (<a href="/products/ET1702-81" style="font-weight: bold;text-decoration: underline;">ET1702-81</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.<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.

    Immunocytochemistry analysis of Daudi cells labeling Human IgM with Rabbit anti-Human IgM antibody (ET1702-81) at 1/100 dilution.

    Cells were fixed in 80% precooled methanol 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-Human IgM antibody (ET1702-81) 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.

    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.

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Citation