P4HB Recombinant Rabbit Monoclonal Antibody [JE54-98]
cat.: ET7110-92
Product Type: Recombinant Rabbit monoclonal IgG, primary antibodies
Species reactivity: Human, Mouse, Rat
Applications: WB, IF-Cell, IHC-P, FC
Clonality: Monoclonal
Clone number: JE54-98
Form: Liquid
Storage condition: Store at +4℃ after thawing. Aliquot store at -20℃. 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: 57 kDa
Isotype: IgG
Immunogen: Recombinant protein within Human P4HB aa 360-508 / 508.
Positive control: Human placenta tissue lysate, THP-1 cell lysate, HepG2, MCF-7, rat kidney tissue, human liver tissue, human placenta tissue, human pancreas tissue, mouse colon tissue, HepG2.
Subcellular location: Endoplasmic reticulum, endoplasmic reticulum lumen, cell membrane, melanosome.
Recommended Dilutions:
  WB
  IF-Cell
  IHC-P
  FC
1:1,000-1:10,000
1:50-1:100
1:50-1:200
1:50-1:100
Uniprot #: SwissProt: P07237 Human | P09103 Mouse | P04785 Rat
Alternative names: Cellular thyroid hormone binding protein Cellular thyroid hormone-binding protein Collagen prolyl 4 hydroxylase beta Disulphide Isomerase DSI EC 5.3.4.1 Endoplasmic reticulum resident protein 59 ER protein 59 ERBA2L ERp59 GIT Gltathione insulin transhydrogenase Glutathione insulin transhydrogenase P4HB P4Hbeta p55 PDI PDIA1 PDIA1_HUMAN PDIR PHDB PO4DB PO4HB Procollagen proline 2 oxoglutarate 4 dioxygenase (proline 4 hydroxylase) beta polypeptide (protein disulfide isomerase associated 1) Procollagen proline 2 oxoglutarate 4 dioxygenase beta subunit PROHB Prolyl 4 hydroxylase beta polypeptide Prolyl 4 hydroxylase beta subunit Prolyl 4 hydroxylase subunit beta Prolyl 4-hydroxylase subunit beta Protein disulfide isomerase associated 1 Protein disulfide isomerase, family A, member 1 Protein disulfide isomerase/oxidoreductase Protein disulfide-isomerase Protocollagen hydroxylase Thbp Thyroid hormone binding protein p55 ......
Images
ET7110-92_1.jpg Fig1: Western blot analysis of P4HB on different lysates with Rabbit anti-P4HB antibody (ET7110-92) at 1/500 dilution.

Lane 1: Human placenta tissue lysate(20 µg/Lane)
Lane 2: THP-1 cell lysate

Lysates/proteins at 10 µg/Lane.

Predicted band size: 57 kDa
Observed band size: 57 kDa

Exposure time: 2 minutes;

8% 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 (ET7110-92) 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 1:5,000 dilution was used for 1 hour at room temperature.
ET7110-92_2.jpg Fig2: Immunocytochemistry analysis of HepG2 cells labeling P4HB with Rabbit anti-P4HB antibody (ET7110-92) at 1/100 dilution.

Cells were fixed in 4% paraformaldehyde for 10 minutes at 37 ℃, permeabilized with 0.05% Triton X-100 in PBS for 20 minutes, and then blocked with 2% negative goat serum for 30 minutes at room temperature. Cells were then incubated with Rabbit anti-P4HB antibody (ET7110-92) at 1/100 dilution in 2% negative goat serum overnight at 4 ℃.Alexa Fluor®488 Goat anti-Rabbit IgG was used as the secondary antibody at 1/1,000 dilution.Nuclear DNA was labelled in blue with DAPI.
ET7110-92_3.jpg Fig3: Immunocytochemistry analysis of MCF-7 cells labeling P4HB with Rabbit anti-P4HB antibody (ET7110-92) at 1/100 dilution.

Cells were fixed in 4% paraformaldehyde for 10 minutes at 37 ℃, permeabilized with 0.05% Triton X-100 in PBS for 20 minutes, and then blocked with 2% negative goat serum for 30 minutes at room temperature. Cells were then incubated with Rabbit anti-P4HB antibody (ET7110-92) at 1/100 dilution in 2% negative goat serum overnight at 4 ℃.Alexa Fluor®488 Goat anti-Rabbit IgG was used as the secondary antibody at 1/1,000 dilution. Nuclear DNA was labelled in blue with DAPI.
ET7110-92_4.jpg Fig4: Immunohistochemical analysis of paraffin-embedded rat kidney tissue with Rabbit anti-P4HB antibody (ET7110-92) at 1/50 dilution.

The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 8.0-8.4)) 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 (ET7110-92) at 1/50 dilution for 0.5 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.
ET7110-92_5.jpg Fig5: Immunohistochemical analysis of paraffin-embedded human liver tissue with Rabbit anti-P4HB antibody (ET7110-92) at 1/200 dilution.

The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 8.0-8.4)) 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 (ET7110-92) at 1/200 dilution for 0.5 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.
ET7110-92_6.jpg Fig6: Immunohistochemical analysis of paraffin-embedded human placenta tissue with Rabbit anti-P4HB antibody (ET7110-92) at 1/50 dilution.

The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 8.0-8.4)) 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 (ET7110-92) at 1/50 dilution for 0.5 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.
ET7110-92_7.jpg Fig7: Immunohistochemical analysis of paraffin-embedded human pancreas tissue with Rabbit anti-P4HB antibody (ET7110-92) at 1/200 dilution.

The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 8.0-8.4)) 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 (ET7110-92) at 1/200 dilution for 0.5 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.
ET7110-92_8.jpg Fig8: Immunohistochemical analysis of paraffin-embedded mouse colon tissue with Rabbit anti-P4HB antibody (ET7110-92) at 1/200 dilution.

The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 8.0-8.4)) 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 (ET7110-92) at 1/200 dilution for 0.5 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.
ET7110-92_9.jpg Fig9: Flow cytometric analysis of P4HB was done on HepG2 cells. The cells were fixed, permeabilized and stained with the primary antibody (ET7110-92, 1/50) (red). After incubation of the primary antibody at room temperature for an hour, the cells were stained with a Alexa Fluor 488-conjugated Goat anti-Rabbit IgG Secondary antibody at 1/1000 dilution for 30 minutes.Unlabelled sample was used as a control (cells without incubation with primary antibody; black).
Note: All products are “FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE”.