Anti-PHIP antibody [EPR27150-82] - BSA and Azide free (ab309963)

Knockout Tested Rabbit Recombinant Monoclonal PHIP antibody. Carrier free. Suitable for WB, IHC-P, ICC/IF, Flow Cyt (Intra), IP and reacts with Human samples.

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Images

Immunoprecipitation - Anti-PHIP antibody [EPR27150-82] - BSA and Azide free (AB309963), expandable thumbnail

Key facts

Isotype: IgG
Host species: Rabbit
Storage buffer: pH: 7.2 - 7.4
Constituents: 100% PBS
Form: Liquid
Clonality: Monoclonal

Immunogen

The exact immunogen used to generate this antibody is proprietary information.

Reactivity data

Select an application

Product promiseTestedExpectedPredictedNot recommended

	ChIP-seq	WB	IHC-P	ICC/IF	Flow Cyt (Intra)	IP
Human	Not recommended	Tested	Tested	Tested	Tested	Tested
Mouse	Not recommended	Not recommended	Not recommended	Not recommended	Not recommended	Not recommended
Rat	Not recommended	Not recommended	Not recommended	Not recommended	Not recommended	Not recommended

Not recommended
Not recommended

Species	Dilution info	Notes
Species Human, Mouse, Rat	Dilution info -	Notes -

Tested
Tested

Species	Dilution info	Notes
Species Human	Dilution info -	Notes -

Not recommended
Not recommended

Species	Dilution info	Notes
Species Mouse, Rat	Dilution info -	Notes -

Tested
Tested

Species	Dilution info	Notes
Species Human	Dilution info -	Notes Perform heat-mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

Not recommended
Not recommended

Species	Dilution info	Notes
Species Mouse, Rat	Dilution info -	Notes -

Tested
Tested

Species	Dilution info	Notes
Species Human	Dilution info -	Notes -

Not recommended
Not recommended

Species	Dilution info	Notes
Species Mouse, Rat	Dilution info -	Notes -

Tested
Tested

Species	Dilution info	Notes
Species Human	Dilution info -	Notes -

Not recommended
Not recommended

Species	Dilution info	Notes
Species Mouse, Rat	Dilution info -	Notes -

Tested
Tested

Species	Dilution info	Notes
Species Human	Dilution info -	Notes -

Not recommended
Not recommended

Species	Dilution info	Notes
Species Mouse, Rat	Dilution info -	Notes -

Target data

See full target information PHIP

Function

Probable regulator of the insulin and insulin-like growth factor signaling pathways. Stimulates cell proliferation through regulation of cyclin transcription and has an anti-apoptotic activity through AKT1 phosphorylation and activation. Plays a role in the regulation of cell morphology and cytoskeletal organization.

Alternative names

DCAF14, WDR11, PHIP, PH-interacting protein, DDB1- and CUL4-associated factor 14, IRS-1 PH domain-binding protein, WD repeat-containing protein 11

Recommended products

Knockout Tested Rabbit Recombinant Monoclonal PHIP antibody. Carrier free. Suitable for WB, IHC-P, ICC/IF, Flow Cyt (Intra), IP and reacts with Human samples.

Key facts

Isotype: IgG
Host species: Rabbit
Storage buffer: pH: 7.2 - 7.4
Constituents: 100% PBS
Form: Liquid
Clonality: Monoclonal
Immunogen: The exact immunogen used to generate this antibody is proprietary information.
Carrier free: Yes
Clone number: EPR27150-82
Purification technique: Affinity purification Protein A
Concentration

Storage

Shipped at conditions: Blue Ice
Appropriate short-term storage conditions: +4°C
Appropriate long-term storage conditions: +4°C

Notes

Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMAb® patents.

This product is a recombinant monoclonal antibody, which offers several advantages including:

- High batch-to-batch consistency and reproducibility
- Improved sensitivity and specificity
- Long-term security of supply
- Animal-free batch production

For more information, read more on recombinant antibodies.

Our carrier-free antibodies are typically supplied in a PBS-only formulation, purified and free of BSA, sodium azide and glycerol. The carrier-free buffer and high concentration allow for increased conjugation efficiency.

This conjugation-ready format is designed for use with fluorochromes, metal isotopes, oligonucleotides, and enzymes, which makes them ideal for antibody labelling, functional and cell-based assays, flow-based assays (e.g. mass cytometry) and Multiplex Imaging applications.

Use our conjugation kits for antibody conjugates that are ready-to-use in as little as 20 minutes with 1 minute hands-on-time and 100% antibody recovery: available for fluorescent dyes, HRP, biotin and gold.

This product is compatible with the Maxpar^® Antibody Labeling Kit from Fluidigm, without the need for antibody preparation. Maxpar^® is a trademark of Fluidigm Canada Inc.

Supplementary info

This supplementary information is collated from multiple sources and compiled automatically.

Activity summary

PHIP also known as pleckstrin homology domain interacting protein is an adapter protein that interacts with insulin receptor substrates. It has a mass of approximately 100 kDa. PHIP is expressed in various tissues including the brain and liver. The protein is involved in intracellular signaling pathways by binding phosphoinositides which are important for cellular communication and growth regulation.

Biological function summary

PHIP plays a significant role in cell proliferation and survival. It contributes to the regulation of the cell cycle by influencing transcription factors and other core components involved in cell growth. PHIP is not only a standalone protein but also part of complexes that modulate chromatin affecting gene expression patterns.

Pathways

PHIP is integrated into insulin signaling and PI3K/AKT pathways. In these pathways PHIP interacts with key proteins like AKT1 supporting cellular responses to insulin and growth factors. The involvement of PHIP in these pathways highlights its role in metabolic regulation and its impact on cell growth and metabolism.

Associated diseases and disorders

PHIP has links to cancer and diabetes. Its dysregulation contributes to abnormal cell proliferation often observed in cancers. PHIP's interaction with insulin signaling components such as IRS1 demonstrates its relevance to diabetes management. These connections show PHIP's potential as a therapeutic target for these conditions.

Product promise

We are dedicated to supporting your work with high quality reagents and we are here for you every step of the way should you need us.

In the unlikely event of one of our products not working as expected, you are covered by our product promise.

Full details and terms and conditions can be found here:
Terms & Conditions.

11 product images

Immunoprecipitation - Anti-PHIP antibody [EPR27150-82] - BSA and Azide free (ab309963)
This data was developed using Anti-PHIP antibody [EPR27150-82] ab309962, the same antibody clone in a different buffer formulation.
PHIP was immunoprecipitated from 0.35 mg HeLa (human cervical adenocarcinoma epithelial cell) whole cell lysate with Anti-PHIP antibody [EPR27150-82] ab309962 at 1/30 dilution (2ug in 0.35mg lysates). Western blot was performed on the immunoprecipitate using Anti-PHIP antibody [EPR27150-82] ab309962 at 1/1000 dilution. VeriBlot for IP secondary antibody(HRP)(VeriBlot for IP Detection Reagent (HRP) ab131366) was used at 1/5000 dilution.
Lane 1: HeLa (human cervical adenocarcinoma epithelial cell) whole cell lysate
Lane 2: Anti-PHIP antibody [EPR27150-82] ab309962 IP in HeLa (human cervical adenocarcinoma epithelial cell) whole cell lysate
Lane 3:Rabbit monoclonal IgG (Rabbit IgG, monoclonal [EPR25A] - Isotype Control ab172730) instead of Anti-PHIP antibody [EPR27150-82] ab309962 in HeLa whole cell lysate
Blocking and dilution buffer and concentration: 5% NFDM/TBST.
Exposure time: 3 minutes
All lanes: Immunoprecipitation - Anti-PHIP antibody [EPR27150-82] (Anti-PHIP antibody [EPR27150-82] ab309962) at 1/30 dilution
All lanes: HeLa (human cervical adenocarcinoma epithelial cell) whole cell lysate
Secondary
All lanes: Immunoprecipitation - VeriBlot for IP Detection Reagent (HRP) (VeriBlot for IP Detection Reagent (HRP) ab131366) at 1/5000 dilution
Exposure time: 3min
Western blot - Anti-PHIP antibody [EPR27150-82] - BSA and Azide free (ab309963)
This data was developed using Anti-PHIP antibody [EPR27150-82] ab309962, the same antibody clone in a different buffer formulation.
Blocking and diluting buffer and concentration: 5% NFDM/TBST
The band beneath the target band (250 kDa) is likely to be degraded target fragment.
In Western blot, anti-Vinculin antibody (Anti-Vinculin antibody [EPR8185] ab129002) loading control staining at 1/10000 dilution.
Exposure time: 48 seconds
All lanes: Western blot - Anti-PHIP antibody [EPR27150-82] (Anti-PHIP antibody [EPR27150-82] ab309962) at 1/1000 dilution
Lane 1: HeLa (human cervical adenocarcinoma epithelial cell) transfected with scrambled siRNA control whole cell lysate at 20 µg
Lane 2: HeLa transfected with siRNA specifically targeti PHIP whole cell lysate at 20 µg
Secondary
All lanes: Western blot - Goat Anti-Rabbit IgG H&L (HRP) (Goat Anti-Rabbit IgG H&L (HRP) ab97051) at 1/20000 dilution
Observed band size: 250 kDa
Exposure time: 48s
Western blot - Anti-PHIP antibody [EPR27150-82] - BSA and Azide free (ab309963)
This data was developed using Anti-PHIP antibody [EPR27150-82] ab309962, the same antibody clone in a different buffer formulation.
Blocking and diluting buffer and concentration: 5% NFDM/TBST
Lysates were freshly made and used for Western blotting immediately to minimize protein degradation.

Exposure time: 59 seconds
All lanes: Western blot - Anti-PHIP antibody [EPR27150-82] (Anti-PHIP antibody [EPR27150-82] ab309962) at 1/1000 dilution
All lanes: 293T (human embryonic kidney epithelial cell) whole cell lysate at 20 µg
Secondary
All lanes: Western blot - Goat Anti-Rabbit IgG H&L (HRP) (Goat Anti-Rabbit IgG H&L (HRP) ab97051) at 1/20000 dilution
Observed band size: 250 kDa
Exposure time: 59s
Western blot - Anti-PHIP antibody [EPR27150-82] - BSA and Azide free (ab309963)
This data was developed using Anti-PHIP antibody [EPR27150-82] ab309962, the same antibody clone in a different buffer formulation.
Blocking and diluting buffer and concentration: 5% NFDM/TBST
Performed under reducing conditions.
In Western blot, Anti-PHIP antibody [EPR27150-82] ab309962 was shown to bind specifically to PHIP. A band was observed at 250 kDa in wild-type Hela cell lysates whereas no signal observed at this size in PHIP knockout cell line Human PHIP knockout HeLa cell line ab265403 (knockout cell lysate Human PHIP knockout HeLa cell lysate ab259039).
The band beneath the target band (250 kDa) is likely to be degraded target fragment.
In Western blot, anti- Vinculin antibody (Anti-Vinculin antibody [EPR8185] ab129002) loading control staining at 1/10000 dilution.
Exposure time: 3 minutes
All lanes: Western blot - Anti-PHIP antibody [EPR27150-82] (Anti-PHIP antibody [EPR27150-82] ab309962) at 1/1000 dilution
Lane 1: Wild-type HeLa(human cervical adenocarcinoma epithelial cell) whole cell lysate at 40 µg
Lane 2: Western blot - Human PHIP knockout HeLa cell lysate (Human PHIP knockout HeLa cell lysate ab259039) at 40 µg
Lane 3: K-562 (human chronic myelogenous leukemia lymphoblast) whole cell lysate at 20 µg
Secondary
All lanes: Western blot - Goat Anti-Rabbit IgG H&L (HRP) (Goat Anti-Rabbit IgG H&L (HRP) ab97051) at 1/20000 dilution
Observed band size: 250 kDa
Exposure time: 3min
Flow Cytometry (Intracellular) - Anti-PHIP antibody [EPR27150-82] - BSA and Azide free (ab309963)
This data was developed using Anti-PHIP antibody [EPR27150-82] ab309962, the same antibody clone in a different buffer formulation.
Flow cytometric analysis of PHIP KO HeLa (PHIP knockout human cervical adenocarcinoma epithelial cell, Left) / Parental HeLa (Right) cells labelling PHIP with Anti-PHIP antibody [EPR27150-82] ab309962 at 1/500 dilution (0.1 ug)/Red compared with a Rabbit monoclonal IgG (Rabbit IgG, monoclonal [EPR25A] - Isotype Control ab172730) (Black) isotype control and an unlabelled control (cells without incubation with primary antibody and secondary antibody) (Blue). Goat Anti-Rabbit IgG (Alexa Fluor® 488, Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed ab150081) at 1/5000 dilution was used as the secondary antibody.
Immunocytochemistry/ Immunofluorescence - Anti-PHIP antibody [EPR27150-82] - BSA and Azide free (ab309963)
This data was developed using Anti-PHIP antibody [EPR27150-82] ab309962, the same antibody clone in a different buffer formulation.
Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% TritonX-100 permeabilized PHIP KO HeLa (human cervical adenocarcinoma epithelial cell) (Human PHIP knockout HeLa cell line ab265403) cells labelling PHIP with Anti-PHIP antibody [EPR27150-82] ab309962 at 1/100 (4.92 ug/ml) dilution, followed by Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed antibody at 1/1000 2 ug/mL dilution (Green). Confocal image showing nuclear staining in parental HeLa cells.Image was taken with a confocal microscope(Leica-Microsystems, TCS SP8). is observed. Alexa Fluor® 594 Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker ab195889 Anti-alpha Tubulin mouse monoclonal antibody - Microtubule Marker (Alexa Fluor® 594) was used to counterstain tubulin at 1/200 2.5ug/ml dilution (Red). The Nuclear counterstain was DAPI (Blue). Secondary antibody only control: Secondary antibody is Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed at 1/1000 2 ug/mL dilution.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-PHIP antibody [EPR27150-82] - BSA and Azide free (ab309963)
This data was developed using Anti-PHIP antibody [EPR27150-82] ab309962, the same antibody clone in a different buffer formulation.
Immunohistochemical analysis of paraffin-embedded (A) Wild-type Hela ( tissue labeling PHIP with Anti-PHIP antibody [EPR27150-82] ab309962 at 1/2000 (0.246 ug/ml) followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection). Nuclear staining on (A) Wild-type Hela cell pellet, no staining on (B) PHIP knockout Hela (Human PHIP knockout HeLa cell line ab265403) cell pellet.The section was incubated with Anti-PHIP antibody [EPR27150-82] ab309962 for 30 mins at room temperature.The immunostaining was performed on a Leica Biosystems BOND® RX instrument Counterstained with Hematoxylin.
Secondary antibody only control: Secondary antibody is a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).
Heat mediated antigen retrieval was performed with Tris-EDTA buffer (pH 9.0, Epitope Retrieval Solution2) for 20 mins
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-PHIP antibody [EPR27150-82] - BSA and Azide free (ab309963)
This data was developed using Anti-PHIP antibody [EPR27150-82] ab309962, the same antibody clone in a different buffer formulation.
Immunohistochemical analysis of paraffin-embedded Human breast carcino tissue labeling PHIP with Anti-PHIP antibody [EPR27150-82] ab309962 at 1/100 (4.92 ug/ml) followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection). Nuclear staining on human breast carcinoma.The section was incubated with Anti-PHIP antibody [EPR27150-82] ab309962 for 30 mins at room temperature.The immunostaining was performed on a Leica Biosystems BOND® RX instrument Counterstained with Hematoxylin.
Secondary antibody only control: Secondary antibody is a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).
Heat mediated antigen retrieval was performed with Tris-EDTA buffer (pH 9.0, Epitope Retrieval Solution2) for 20 mins
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-PHIP antibody [EPR27150-82] - BSA and Azide free (ab309963)
This data was developed using Anti-PHIP antibody [EPR27150-82] ab309962, the same antibody clone in a different buffer formulation.
Immunohistochemical analysis of paraffin-embedded Human lung adenocarc tissue labeling PHIP with Anti-PHIP antibody [EPR27150-82] ab309962 at 1/100 (4.92 ug/ml) followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection). Nuclear staining on human lung adenocarcinoma.The section was incubated with Anti-PHIP antibody [EPR27150-82] ab309962 for 30 mins at room temperature.The immunostaining was performed on a Leica Biosystems BOND® RX instrument Counterstained with Hematoxylin.
Secondary antibody only control: Secondary antibody is a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).
Heat mediated antigen retrieval was performed with Tris-EDTA buffer (pH 9.0, Epitope Retrieval Solution2) for 20 mins
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-PHIP antibody [EPR27150-82] - BSA and Azide free (ab309963)
This data was developed using Anti-PHIP antibody [EPR27150-82] ab309962, the same antibody clone in a different buffer formulation.
Immunohistochemical analysis of paraffin-embedded Human colon tissue labeling PHIP with Anti-PHIP antibody [EPR27150-82] ab309962 at 1/100 (4.92 ug/ml) followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection). Nuclear staining on human colon.The section was incubated with Anti-PHIP antibody [EPR27150-82] ab309962 for 30 mins at room temperature.The immunostaining was performed on a Leica Biosystems BOND® RX instrument Counterstained with Hematoxylin.
Secondary antibody only control: Secondary antibody is a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).
Heat mediated antigen retrieval was performed with Tris-EDTA buffer (pH 9.0, Epitope Retrieval Solution2) for 20 mins
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-PHIP antibody [EPR27150-82] - BSA and Azide free (ab309963)
This data was developed using Anti-PHIP antibody [EPR27150-82] ab309962, the same antibody clone in a different buffer formulation.
Immunohistochemical analysis of paraffin-embedded Human pancreas tissue labeling PHIP with Anti-PHIP antibody [EPR27150-82] ab309962 at 1/100 (4.92 ug/ml) followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection). Nuclear staining on human pancreas.The section was incubated with Anti-PHIP antibody [EPR27150-82] ab309962 for 30 mins at room temperature.The immunostaining was performed on a Leica Biosystems BOND® RX instrument Counterstained with Hematoxylin.
Secondary antibody only control: Secondary antibody is a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).
Heat mediated antigen retrieval was performed with Tris-EDTA buffer (pH 9.0, Epitope Retrieval Solution2) for 20 mins