CA125/MUC16 Recombinant Rabbit Monoclonal Antibody [PSH21-77]
cat.: HA724248
Product Type: Recombinant Rabbit monoclonal IgG, primary antibodies
Species reactivity: Human
Applications: WB, IF-Cell, IHC-P, FC, IP
Clonality: Monoclonal
Clone number: PSH21-77
Form: Liquid
Storage condition: 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: 1*PBS (pH7.4), 0.1% BSA, 40% Glycerol, 0.2% Proclean 950.
Concentration: 1ug/ul
Purification: Protein A affinity purified.
Molecular weight: Predicted band size: 1519 kDa
Isotype: IgG
Immunogen: Recombinant protein within human CA125/MUC16 aa 13,302-14,451.
Positive control: NIH:OVCAR-3 cell lysate, HeLa cell lysate, human ovarian carcinoma tissue, human salivary glands tissue, NIH:OVCAR-3.
Subcellular location: Cell membrane, secreted, extracellular space.
Recommended Dilutions:
  WB
  IF-Cell
  IHC-P
  FC
  IP
1:5,000
1:500
1:200-1:1,000
1:1,000
1-2μg/sample
Uniprot #: SwissProt: Q8WXI7 Human
Alternative names: CA 125 CA-125 CA125 CA125 ovarian cancer antigen Cancer antigen 125 FLJ14303 MUC 16 MUC-16 MUC16 MUC16_HUMAN Mucin 16 Mucin 16 cell surface associated Mucin-16 Mucin16 Ovarian cancer related tumor marker CA125 Ovarian cancer-related tumor marker CA125 Ovarian carcinoma antigen CA125
Images
HA724248_1.jpg Fig1: Western blot analysis of CA125/MUC16 on different lysates with Rabbit anti-CA125/MUC16 antibody (HA724248) at 1/5,000 dilution.

Lane 1: NIH:OVCAR-3 cell lysate
Lane 2: SK-OV-3 cell lysate (negative)
Lane 3: HeLa cell lysate
Lane 4: MCF7 cell lysate (negative)

Lysates/proteins at 15 µg/Lane.

Predicted band size: 1519 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 (HA724248) at 1/5,000 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.
HA724248_2.jpg Fig2: Immunocytochemistry analysis of NIH:OVCAR-3 (positive) and SK-OV-3 (negative) labeling CA125/MUC16 with Rabbit anti-CA125/MUC16 antibody (HA724248) at 1/500 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-CA125/MUC16 antibody (HA724248) at 1/500 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.
HA724248_3.jpg Fig3: Immunohistochemical analysis of paraffin-embedded human ovarian carcinoma tissue with Rabbit anti-CA125/MUC16 antibody (HA724248) 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 (HA724248) 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.
HA724248_4.jpg Fig4: Immunohistochemical analysis of paraffin-embedded human ovarian carcinoma tissue with Rabbit anti-CA125/MUC16 antibody (HA724248) 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 (HA724248) 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.
HA724248_5.jpg Fig5: Immunohistochemical analysis of paraffin-embedded human ovarian carcinoma tissue with Rabbit anti-CA125/MUC16 antibody (HA724248) 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 (HA724248) 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.
HA724248_6.jpg Fig6: Immunohistochemical analysis of paraffin-embedded human salivary glands tissue with Rabbit anti-CA125/MUC16 antibody (HA724248) at 1/200 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 (HA724248) at 1/200 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.
HA724248_7.jpg Fig7: Immunohistochemical analysis of paraffin-embedded human hepatocellular carcinoma tissue with Rabbit anti-CA125/MUC16 antibody (HA724248) at 1/200 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 (HA724248) at 1/200 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.
HA724248_8.jpg Fig8: Flow cytometric analysis of SK-OV-3 (left, negative) and NIH:OVCAR-3 (right, positive) cells labeling CA125/MUC16.

Cells were washed twice with cold PBS and resuspend. Then stained with the primary antibody (HA724248, 1/1,000) (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).
HA724248_9.jpg Fig9: CA125/MUC16 was immunoprecipitated from 0.2 mg NIH:OVCAR-3 cell lysate with HA724248 at 2 µg/10 µl beads. Western blot was performed from the immunoprecipitate using HA724248 at 1/5,000 dilution. HRP Conjugated Anti-Rabbit IgG for IP Nano-secondary antibody at 1/5,000 dilution was used for 1 hour at room temperature.

Lane 1: NIH:OVCAR-3 cell lysate (input)
Lane 2: HA724248 IP in NIH:OVCAR-3 cell lysate
Lane 3: Rabbit IgG instead of HA724248 in NIH:OVCAR-3 cell lysate

Blocking/Dilution buffer: primary antibody dilution (K1803)
Exposure time: 3 seconds; ECL: K1801
Note: All products are “FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE”.