WB
Western blot analysis of HIF Prolyl Hydroxylases on different lysates with Rabbit anti-HIF Prolyl Hydroxylases antibody at 1/1,000 dilution. Lysates/proteins at 20 µg/Lane. Exposure time: 25 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 at 1/1,000 dilution was used in 5% NFDM/TBST at 4℃ overnight. Goat Anti-Rabbit IgG - HRP Secondary Antibody at 1/50,000 dilution was used for 1 hour at room temperature.IHC
Immunohistochemical analysis of paraffin-embedded human lung cancer tissue with Rabbit anti-HIF Prolyl Hydroxylases antibody 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 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.ICC/IF
Immunocytochemistry analysis of HEK-293 cells labeling HIF Prolyl Hydroxylases with Rabbit anti-HIF Prolyl Hydroxylases antibody 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-HIF Prolyl Hydroxylases antibody at 1/100 dilution in 1% BSA in PBST overnight at 4 ℃. Goat Anti-Rabbit IgG H&L (488) 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 (red) was stained at 1/100 dilution overnight at +4℃. Goat Anti-Mouse IgG H&L (594) was used as the secondary antibody at 1/1,000 dilution.FC
Flow cytometric analysis of HEK-293 cells labeling HIF Prolyl Hydroxylases. Cells were fixed and permeabilized. Then stained with the primary antibody (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 488 conjugate-Goat anti-Rabbit IgG Secondary antibody at 1/1,000 dilution for 30 minutes at +4℃. Unlabelled sample was used as a control (cells without incubation with primary antibody; black).IP
HIF Prolyl Hydroxylases was immunoprecipitated from 0.2 mg HEK-293 cell lysate with HIF Prolyl Hydroxylases Recombinant Rabbit Monoclonal Antibody at 2 µg/10 µl beads. Western blot was performed from the immunoprecipitate using HIF Prolyl Hydroxylases Recombinant Rabbit Monoclonal Antibody at 1/1,000 dilution. Anti-Rabbit IgG for IP, VHH(HRP) at 1/5,000 dilution was used for 1 hour at room temperature. Lane 1: HEK-293 cell lysate (input). Lane 2: HIF Prolyl Hydroxylases Recombinant Rabbit Monoclonal Antibody IP in HEK-293 cell lysate. Lane 3: Rabbit IgG instead of HIF Prolyl Hydroxylases Recombinant Rabbit Monoclonal Antibody in HEK-293 cell lysate. Blocking/Dilution buffer: 5% NFDM/TBST. Exposure time: 26 seconds.| Product Name | HIF Prolyl Hydroxylases Recombinant Rabbit Monoclonal Antibody |
|---|---|
| Antibody Type | Primary Antibodies |
| Immunogen | Synthetic peptide within Human HIF Prolyl Hydroxylases aa 51-150 / 502. |
| Clonality | Monoclonal |
|---|---|
| Isotype | IgG |
| Host Species | Rabbit |
| Tested Applications | FCICC/IFIHCIPWB |
| WB:1:1000 IHC:1:200 ICC/IF:1:100 FC:1:1000 IP:1-2μg/sample |
|
| Species Reactivity | Human |
| Concentration | 1mg/ml |
| Purification | Protein A |
| Gene Symbol | P4HTM |
|---|---|
| Gene Synonyms | PH4 PH-4 PHD4 EGLN4 HIDEA HIFPH4 P4H-TM |
| Gene Full Name | prolyl 4-hydroxylase, transmembrane |
| Gene Summary | The product of this gene belongs to the family of prolyl 4-hydroxylases. This protein is a prolyl hydroxylase that may be involved in the degradation of hypoxia-inducible transcription factors under normoxia. It plays a role in adaptation to hypoxia and may be related to cellular oxygen sensing. Alternatively spliced variants encoding different isoforms have been identified. [provided by RefSeq, Jul 2008] |
| Alternative Names | EGLN4 antibody FLJ20262 antibody HIF prolyl hydroxylase PH4 antibody HIF-PH4 antibody HIF-prolyl hydroxylase 4 antibody HIFPH4 antibody HPH-4 antibody Hypoxia inducible factor prolyl 4 hydroxylase antibody Hypoxia inducible factor prolyl hydroxylase antibody Hypoxia-inducible factor prolyl hydroxylase 4 antibody P4H TM antibody P4H with transmembrane domain antibody P4H-TM antibody P4htm antibody P4HTM_HUMAN antibody PH 4 antibody PH4 antibody PHD4 antibody Proline 4 hydroxylase antibody Prolyl 4 hydroxylase transmembrane (endoplasmic reticulum) antibody Prolyl hydroxlase domain containing 4 antibody Transmembrane prolyl 4-hydroxylase antibody |
| Molecular Weight(MW) | 47 kDa |
| Cellular Localization | Endoplasmic reticulum,membrane. |
| SwissProt ID | Q9NXG6 |
|---|
WB
Western blot analysis of HIF Prolyl Hydroxylases on different lysates with Rabbit anti-HIF Prolyl Hydroxylases antibody at 1/1,000 dilution. Lysates/proteins at 20 µg/Lane. Exposure time: 25 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 at 1/1,000 dilution was used in 5% NFDM/TBST at 4℃ overnight. Goat Anti-Rabbit IgG - HRP Secondary Antibody at 1/50,000 dilution was used for 1 hour at room temperature.
IHC
Immunohistochemical analysis of paraffin-embedded human lung cancer tissue with Rabbit anti-HIF Prolyl Hydroxylases antibody 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 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.
ICC/IF
Immunocytochemistry analysis of HEK-293 cells labeling HIF Prolyl Hydroxylases with Rabbit anti-HIF Prolyl Hydroxylases antibody 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-HIF Prolyl Hydroxylases antibody at 1/100 dilution in 1% BSA in PBST overnight at 4 ℃. Goat Anti-Rabbit IgG H&L (488) 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 (red) was stained at 1/100 dilution overnight at +4℃. Goat Anti-Mouse IgG H&L (594) was used as the secondary antibody at 1/1,000 dilution.
FC
Flow cytometric analysis of HEK-293 cells labeling HIF Prolyl Hydroxylases. Cells were fixed and permeabilized. Then stained with the primary antibody (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 488 conjugate-Goat anti-Rabbit IgG Secondary antibody at 1/1,000 dilution for 30 minutes at +4℃. Unlabelled sample was used as a control (cells without incubation with primary antibody; black).
IP
HIF Prolyl Hydroxylases was immunoprecipitated from 0.2 mg HEK-293 cell lysate with HIF Prolyl Hydroxylases Recombinant Rabbit Monoclonal Antibody at 2 µg/10 µl beads. Western blot was performed from the immunoprecipitate using HIF Prolyl Hydroxylases Recombinant Rabbit Monoclonal Antibody at 1/1,000 dilution. Anti-Rabbit IgG for IP, VHH(HRP) at 1/5,000 dilution was used for 1 hour at room temperature. Lane 1: HEK-293 cell lysate (input). Lane 2: HIF Prolyl Hydroxylases Recombinant Rabbit Monoclonal Antibody IP in HEK-293 cell lysate. Lane 3: Rabbit IgG instead of HIF Prolyl Hydroxylases Recombinant Rabbit Monoclonal Antibody in HEK-293 cell lysate. Blocking/Dilution buffer: 5% NFDM/TBST. Exposure time: 26 seconds.| Application Notes | WB:1:1000 IHC:1:200 ICC/IF:1:100 FC:1:1000 IP:1-2μg/sample |
|---|
| Form | Liquid |
|---|---|
| Storage Instructions | Shipped at 4℃. Store at +4℃ short term (1-2 weeks). Store at -20℃ long term. |
| Storage Buffer | 1*TBS (pH7.4), 0.05% BSA, 40% Glycerol. Preservative: 0.05% Sodium Azide. |
