HA721721

PSMA Recombinant Rabbit Monoclonal Antibody [PD00-72]

cat.: HA721721

Product Type:	Recombinant Rabbit monoclonal IgG, primary antibodies
Species reactivity:	Human
Applications:	WB, IHC, IF-Tissue
Clonality:	Monoclonal
Clone number:	PD00-72

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: 84 kDa
Isotype:	IgG
Immunogen:	Synthetic peptide within human PSMA.
Positive control:	LNCaP cell lysate, 22RV1 cell lysate, LNCaP, human prostate cancer tissue, human kidney tissue.
Subcellular location:	Cell membrane, Cytoplasm
Recommended Dilutions: WB IHC IF-Tissue	1:1,000 1:1,000 1:200
Uniprot #:	SwissProt: Q04609 Human
Alternative names:	Cell growth inhibiting protein 27 Cell growth-inhibiting gene 27 protein FGCP Folate hydrolase (prostate-specific membrane antigen) 1 Folate hydrolase 1 Folate hydrolase Folate hydrolase prostate specific membrane antigen 1 FOLH 1 FOLH Folh1 FOLH1_HUMAN Folylpoly gamma glutamate carboxypeptidase Folylpoly-gamma-glutamate carboxypeptidase GCP 2 GCP II GCP2 GCPII GIG27 Glutamate carboxylase II Glutamate carboxypeptidase 2 Glutamate carboxypeptidase II Membrane glutamate carboxypeptidase mGCP N acetylated alpha linked acidic dipeptidase 1 N-acetylated-alpha-linked acidic dipeptidase I NAALAD 1 NAALAD1 NAALAdase NAALADase I Prostate specific membrane antigen Prostate specific membrane antigen variant F Prostate-specific membrane antigen PSM PSMA Pteroylpoly gamma glutamate carboxypeptidase Pteroylpoly-gamma-glutamate carboxypeptidase

Images

Fig1: Western blot analysis of PSMA on different lysates with Rabbit anti-PSMA antibody (HA721721) at 1/1,000 dilution.

Lane 1: LNCaP cell lysate
Lane 2: 22RV1 cell lysate
Lane 3: PC-3M cell lysate (negative)

Lysates/proteins at 20 µg/Lane.

Predicted band size: 84 kDa
Observed band size: 100/200 kDa

Exposure time: 15 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 (HA721721) 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.

Fig2: Immunocytochemistry analysis of LNCaP (positive) and PC-3M (negative) cells labeling PSMA with Rabbit anti-PSMA antibody (HA721721) at 1/100 dilution.

Cells were fixed in 100% 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-PSMA antibody (HA721721) 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: Immunohistochemical analysis of paraffin-embedded human prostate cancer tissue with Rabbit anti-PSMA antibody (HA721721) 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 (HA721721) 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.
	Fig4: Immunohistochemical analysis of paraffin-embedded human kidney tissue with Rabbit anti-PSMA antibody (HA721721) 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 (HA721721) 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 colon tissue (negative control) with Rabbit anti-PSMA antibody (HA721721) 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 (HA721721) 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 breast carcinoma tissue (negative control) with Rabbit anti-PSMA antibody (HA721721) 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 (HA721721) 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: Immunofluorescence analysis of paraffin-embedded human prostate cancer tissue labeling PSMA with Rabbit anti-PSMA antibody (HA721721) 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 10% negative goat serum for 1 hour at room temperature, washed with PBS, and then probed with the primary antibody (HA721721, green) at 1/200 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).

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