Human IgM Recombinant Rabbit Monoclonal Antibody [JF84-09]
cat.: ET1702-81
Product Type: Recombinant Rabbit monoclonal IgG, primary antibodies
Species reactivity: Human
Applications: WB, IHC-P, IP, FC, IF-Cell
Clonality: Monoclonal
Clone number: JF84-09
Form: Liquid
Storage condition: Store at +4℃ after thawing. Aliquot store at -20℃ or -80℃. Avoid repeated freeze / thaw cycles.
Storage buffer: 1*TBS (pH7.4), 0.05% BSA, 40% Glycerol. Preservative: 0.05% Sodium Azide.
Concentration: 1ug/ul
Purification: Protein A affinity purified.
Molecular weight: Predicted band size: 50 kDa
Isotype: IgG
Immunogen: Recombinant protein within Human IgM from Human serum (Sigma#I8260).
Positive control: Daudi cell lysate, human IgM protein, human tonsil tissue, human spleen tissue, Daudi.
Subcellular location: Secreted, Cell membrane.
Recommended Dilutions:
  WB
  IHC-P
  FC
  IF-Cell
1:1,000-1:5,000
1:50-1:200
1:1,000
1:100
Uniprot #: SwissProt: P01871 Human
Alternative names: Immunoglobin heavy chain constant region mu edit item name - Immunoglobin heavy chain mu constant region Immunoglobin heavy chain constant region mu AGM1 Constant region of heavy chain of IgM DKFZp686I15196 DKFZp686I15212 FLJ00385 Ig mu chain C region IGHM IgM heavy chain constant region Immunoglobin heavy constant mu Immunoglobulin mu MGC104996 MGC52291 MU VH
Images
ET1702-81_1.jpg Fig1: 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.
ET1702-81_2.jpg Fig2: Western blot analysis of Human IgM on human IgM protein with Rabbit anti-Human IgM antibody (ET1702-81) at 1/500 dilution.

Lysates/proteins at 50 ng/Lane.

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

Exposure time: 30 seconds;

10% 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 1/500 dilution was used in 5% NFDM/TBST at room temperature for 2 hours. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1:200,000 dilution was used for 1 hour at room temperature.
ET1702-81_3.jpg Fig3: 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.
ET1702-81_4.jpg Fig4: 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.
ET1702-81_5.jpg Fig5: 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).
ET1702-81_6.jpg Fig6: 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.
Note: All products are “FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE”.