HA721810

Mesothelin Recombinant Rabbit Monoclonal Antibody [PSH02-30]

cat.: HA721810

Product Type:	Recombinant Rabbit monoclonal IgG, primary antibodies
Species reactivity:	Human
Applications:	WB, IF-Cell, IF-Tissue, IHC-P
Clonality:	Monoclonal
Clone number:	PSH02-30

Form:	Liquid
Storage condition:	Store at +4℃ after thawing. Aliquot store at -20℃. Avoid repeated freeze / thaw cycles.
Storage buffer:	PBS (pH7.4), 0.1% BSA, 40% Glycerol. Preservative: 0.05% Sodium Azide.
Concentration:	1ug/ul
Purification:	Protein A affinity purified.
Molecular weight:	Predicted band size: 68 kDa
Isotype:	IgG
Immunogen:	Recombinant protein within human Mesothelin aa 273-622 / 622 (Q13421-3).
Positive control:	SiHa cell lysate, NCI-H226 cell lysate, HeLa cell lysate, NIH:OVCAR-3 cell lysate, NCI-H226, human ovarian carcinoma tissue, human tonsil tissue.
Subcellular location:	Cell membrane, Golgi apparatus; Secreted.
Recommended Dilutions: WB IF-Cell IF-Tissue IHC-P	1:1,000 1:100 1:100 1:1,000
Uniprot #:	SwissProt: Q13421 Human
Alternative names:	CAK 1 CAK1 CAK1 antigen cleaved form Megakaryocyte potentiating factor Mesothelin Mesothelin isoform 1 precursor MPF Msln MSLN_HUMAN Pre pro megakaryocyte potentiating factor Pre-pro-megakaryocyte-potentiating factor SMR SMRP Soluble MPF mesothelin related protein

Images

Fig1: Western blot analysis of Mesothelin on different lysates with Rabbit anti-Mesothelin antibody (HA721810) at 1/1,000 dilution.

Lane 1: SiHa cell lysate
Lane 2: NCI-H226 cell lysate
Lane 3: HeLa cell lysate
Lane 4: NIH:OVCAR-3 cell lysate
Lane 5: PC-3M cell lysate (negative)
Lane 6: A549 cell lysate (negative)

Lysates/proteins at 20 µg/Lane.

Predicted band size: 68 kDa
Observed band size: 40 kDa

Exposure time: 1 minute 2 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 (HA721810) at 1/1,000 dilution was used in 5% NFDM/TBST at room temperature for 2 hours. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1/50,000 dilution was used for 1 hour at room temperature.

Fig2: Immunocytochemistry analysis of NCI-H226 cells labeling Mesothelin with Rabbit anti-Mesothelin antibody (HA721810) at 1/100 dilution.

Cells were fixed in 4% paraformaldehyde for 20 minutes at room temperature, permeabilized with 0.1% Triton X-100 in PBS 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-Mesothelin antibody (HA721810) 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.

	Fig3: Immunofluorescence analysis of paraffin-embedded human ovarian carcinoma tissue labeling Mesothelin with Rabbit anti-Mesothelin antibody (HA721810) at 1/100 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 10% negative goat serum for 1 hour at room temperature, washed with PBS, and then probed with the primary antibody (HA721810, green) at 1/100 dilution overnight at 4 ℃, washed with PBS. Goat Anti-Rabbit IgG H&L (iFluor™ 488, HA1121) was used as the secondary antibody at 1/1,000 dilution. Nuclei were counterstained with DAPI (blue).
	Fig4: Immunohistochemical analysis of paraffin-embedded human ovarian carcinoma tissue with Rabbit anti-Mesothelin antibody (HA721810) 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 (HA721810) 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.
	Fig5: Immunohistochemical analysis of paraffin-embedded human tonsil tissue with Rabbit anti-Mesothelin antibody (HA721810) 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 (HA721810) 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.

Fig6: Immunohistochemical analysis of paraffin-embedded human colon tissue (negative control) with Rabbit anti-Mesothelin antibody (HA721810) 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 (HA721810) 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.

Fig7: Immunohistochemical analysis of paraffin-embedded human gallbladder tissue (negative control) with Rabbit anti-Mesothelin antibody (HA721810) 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 (HA721810) 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.

Note: All products are “FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE”.