Anti-PHC2 antibody [EPR27147-176] - ChIP Grade - BSA and Azide free
- BOND RX™ Validated
- Recombinant
- Advanced Validation
- RabMAb
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Rabbit Recombinant Monoclonal PHC2 antibody. Carrier free. Suitable for ChIP-seq, WB, ICC/IF, IP, Flow Cyt (Intra), IHC-P and reacts with Human, Mouse, Rat, Recombinant fragment - Human samples.
View Alternative Names
EDR2, PH2, PHC2, Polyhomeotic-like protein 2, hPH2, Early development regulatory protein 2
- ICC/IF
Supplier Data
Immunocytochemistry/ Immunofluorescence - Anti-PHC2 antibody [EPR27147-176] - ChIP Grade - BSA and Azide free (AB314437)
This data was developed using ab314436, the same antibody clone in a different buffer formulation.Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% TritonX-100 permeabilized SW480 (human colorectal adenocarcinoma epithelial cell) cells labelling PHC2 with ab314436 at 1/50 (10.38 ug/ml) dilution, followed by 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 SW480 cell line.Image was taken with a confocal microscope(Leica-Microsystems, TCS SP8).ab195889 Anti-alpha Tubulin mouse monoclonal antibody - Microtubule Marker (Alexa Fluor® 594) was used to counterstain tubulin at 1/200 (2.5 ug/ml) dilution (Red). The Nuclear counterstain was DAPI (Blue). Secondary antibody only control : Secondary antibody is ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed at 1/1000 (2 ug/ml) dilution.
- IHC-P
Supplier Data
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-PHC2 antibody [EPR27147-176] - ChIP Grade - BSA and Azide free (AB314437)
This data was developed using ab314436, the same antibody clone in a different buffer formulation. Immunohistochemical analysis of paraffin-embedded human cerebrum tissue labeling PHC2 with ab314436 at 1/100 (5.19 ug/ml) followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection). Nuclear staining on human cerebrum. The section was incubated with ab314436 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.
- IHC-P
Supplier Data
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-PHC2 antibody [EPR27147-176] - ChIP Grade - BSA and Azide free (AB314437)
This data was developed using ab314436, the same antibody clone in a different buffer formulation. Immunohistochemical analysis of paraffin-embedded human kidney tissue labeling PHC2 with ab314436 at 1/100 (5.19 ug/ml) followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection). Nuclear staining on human kidney.The section was incubated with ab314436 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.
- ChIP-seq
Lab
ChIP-sequencing - Anti-PHC2 antibody [EPR27147-176] - ChIP Grade - BSA and Azide free (AB314437)
Chromatin was prepared from NCCIT cells. Cells were fixed with 1% formaldehyde for 10 minutes. ChIP was performed with 10^7 cells and 8 µg of ab314436 [EPR27147-176]. ChIP DNA was sequenced on the Illumina NovaSeq 6000 to a depth of 30 million reads. The Input control is also shown.
- ChIP-seq
Lab
ChIP-sequencing - Anti-PHC2 antibody [EPR27147-176] - ChIP Grade - BSA and Azide free (AB314437)
Chromatin was prepared from NCCIT cells. Cells were fixed with 1% formaldehyde for 10 minutes. ChIP was performed with 10^7 cells and 8 µg of ab314436 [EPR27147-176]. ChIP DNA was sequenced on the Illumina NovaSeq 6000 to a depth of 30 million reads. The Input control is also shown. This data was developed using ab314436, the same antibody clone in a different buffer formulation.
- ChIP-seq
Lab
ChIP-sequencing - Anti-PHC2 antibody [EPR27147-176] - ChIP Grade - BSA and Azide free (AB314437)
Chromatin was prepared from NCCIT cells. Cells were fixed with 1% formaldehyde for 10 minutes. ChIP was performed with 10^7 cells and 8 µg of ab314436 [EPR27147-176]. ChIP DNA was sequenced on the Illumina NovaSeq 6000 to a depth of 30 million reads. The Input control is also shown. This data was developed using ab314436, the same antibody clone in a different buffer formulation.
- IP
Supplier Data
Immunoprecipitation - Anti-PHC2 antibody [EPR27147-176] - ChIP Grade - BSA and Azide free (AB314437)
This data was developed using ab314436, the same antibody clone in a different buffer formulation. PHC2 was immunoprecipitated from 0.35 mg SW480 (human colorectal adenocarcinoma epithelial cell) whole cell lysate with ab314436 at 1/30 dilution (2ug in 0.35mg lysates). Western blot was performed on the immunoprecipitate using ab314436 at 1/1000 dilution. VeriBlot for IP secondary antibody(HRP)(ab131366) was used at 1/5000 dilution. Lane 1 : SW480 (human colorectal adenocarcinoma epithelial cell) whole cell lysate Lane 2 : ab314436 IP in SW480 (human colorectal adenocarcinoma epithelial cell) whole cell lysate Lane 3 : Rabbit monoclonal IgG (ab172730) instead of ab314436 in SW480 whole cell lysate Blocking and dilution buffer and concentration : 5% NFDM/TBST.
All lanes:
Immunoprecipitation - Anti-PHC2 antibody [EPR27147-176] - ChIP Grade (<a href='/en-us/products/primary-antibodies/phc2-antibody-epr27147-176-chip-grade-ab314436'>ab314436</a>) at 1/30 dilution
All lanes:
SW480 (human colorectal adenocarcinoma epithelial cell) whole cell lysate
Secondary
All lanes:
Immunoprecipitation - VeriBlot for IP Detection Reagent (HRP) (<a href='/en-us/products/reagents/veriblot-for-ip-detection-reagent-hrp-ab131366'>ab131366</a>) at 1/5000 dilution
false
Exposure time: 58s
- IHC-P
Supplier Data
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-PHC2 antibody [EPR27147-176] - ChIP Grade - BSA and Azide free (AB314437)
This data was developed using ab314436, the same antibody clone in a different buffer formulation. Immunohistochemical analysis of paraffin-embedded rat cerebrum tissue labeling PHC2 with ab314436 at 1/100 (5.19 ug/ml) followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection). Nuclear staining on rat cerebrum. The section was incubated with ab314436 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.
- IHC-P
Supplier Data
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-PHC2 antibody [EPR27147-176] - ChIP Grade - BSA and Azide free (AB314437)
This data was developed using ab314436, the same antibody clone in a different buffer formulation. Immunohistochemical analysis of paraffin-embedded rat kidney tissue labeling PHC2 with ab314436 at 1/100 (5.19 ug/ml) followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection). Nuclear staining on rat kidney. The section was incubated with ab314436 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.
- IHC-P
Supplier Data
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-PHC2 antibody [EPR27147-176] - ChIP Grade - BSA and Azide free (AB314437)
This data was developed using ab314436, the same antibody clone in a different buffer formulation. Immunohistochemical analysis of paraffin-embedded mouse kidney tissue labeling PHC2 with ab314436 at 1/100 (5.19 ug/ml) followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection). Nuclear staining on mouse kidney. The section was incubated with ab314436 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.
- ICC/IF
Supplier Data
Immunocytochemistry/ Immunofluorescence - Anti-PHC2 antibody [EPR27147-176] - ChIP Grade - BSA and Azide free (AB314437)
This data was developed using ab314436, the same antibody clone in a different buffer formulation.Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% TritonX-100 permeabilized NIH/3T3 (mouse embryonic fibroblast) cells labelling PHC2 with ab314436 at 1/50 (10.38 ug/ml) dilution, followed by 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 NIH/3T3 cell line.Image was taken with a confocal microscope(Leica-Microsystems, TCS SP8). ab195889 Anti-alpha Tubulin mouse monoclonal antibody - Microtubule Marker (Alexa Fluor® 594) was used to counterstain tubulin at 1/200 (2.5 ug/ml) dilution (Red). The Nuclear counterstain was DAPI (Blue). Secondary antibody only control : Secondary antibody is ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed at 1/1000 (2 ug/ml) dilution.
- Flow Cyt (Intra)
Supplier Data
Flow Cytometry (Intracellular) - Anti-PHC2 antibody [EPR27147-176] - ChIP Grade - BSA and Azide free (AB314437)
This data was developed using ab314436, the same antibody clone in a different buffer formulation.Flow cytometric analysis of 4% paraformaldehyde fixed 90% methanol permeabilized NIH/3T3 (mouse embryonic fibroblast) cells labelling PHC2 with ab314436 at 1/50 dilution (1 ug)/Red (Red) compared with a Rabbit monoclonal IgG (ab172730) (Black) isotype control and an unlabelled control (cells without incubation with primary antibody and secondary antibody) (Blue). A Goat Anti-Rabbit IgG (Alexa Fluor® 488, ab150081) at 1/5000 dilution was used as the secondary antibody.
- IHC-P
Supplier Data
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-PHC2 antibody [EPR27147-176] - ChIP Grade - BSA and Azide free (AB314437)
This data was developed using ab314436, the same antibody clone in a different buffer formulation. Immunohistochemical analysis of paraffin-embedded mouse cerebrum tissue labeling PHC2 with ab314436 at 1/100 (5.19 ug/ml) followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection). Nuclear staining on mouse cerebrum. The section was incubated with ab314436 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.
- IP
Supplier Data
Immunoprecipitation - Anti-PHC2 antibody [EPR27147-176] - ChIP Grade - BSA and Azide free (AB314437)
This data was developed using ab314436, the same antibody clone in a different buffer formulation. PHC2 was immunoprecipitated from 0.35 mg NIH/3T3 (mouse embryonic fibroblast) whole cell lysate with ab314436 at 1/30 dilution (2ug in 0.35mg lysates). Western blot was performed on the immunoprecipitate using ab314436 at 1/1000 dilution. VeriBlot for IP secondary antibody(HRP)(ab131366) was used at 1/5000 dilution. Lane 1 : NIH/3T3 (mouse embryonic fibroblast) whole cell lysate Lane 2 : ab314436 IP in NIH/3T3 (mouse embryonic fibroblast) whole cell lysate Lane 3 : Rabbit monoclonal IgG (ab172730) instead of ab314436 NIH/3T3 whole cell lysate Blocking and dilution buffer and concentration : 5% NFDM/TBST.
All lanes:
Immunoprecipitation - Anti-PHC2 antibody [EPR27147-176] - ChIP Grade (<a href='/en-us/products/primary-antibodies/phc2-antibody-epr27147-176-chip-grade-ab314436'>ab314436</a>) at 1/30 dilution
All lanes:
NIH/3T3 (mouse embryonic fibroblast) whole cell lysate
Secondary
All lanes:
Immunoprecipitation - VeriBlot for IP Detection Reagent (HRP) (<a href='/en-us/products/reagents/veriblot-for-ip-detection-reagent-hrp-ab131366'>ab131366</a>) at 1/5000 dilution
false
Exposure time: 76s
- WB
Supplier Data
Western blot - Anti-PHC2 antibody [EPR27147-176] - ChIP Grade - BSA and Azide free (AB314437)
This data was developed using ab314436, the same antibody clone in a different buffer formulation. Blocking and diluting buffer and concentration : 5% NFDM/TBST The expression profile/molecular weight observed is consistent with what has been described in the literature (PMID : 16024804). The identity of the bands below 31 kDa is unknown. In Western blot, Anti-GAPDH antibody [EPR16891] - Loading Control (ab181602) staining at 1/200000 dilution. In Western blot, Anti-Vinculin antibody [EPR8185] (ab129002) staining at 1/10000 dilution. Exposure time : Lane1 : 180 seconds, lane2-7 : 37 seconds
All lanes:
Western blot - Anti-PHC2 antibody [EPR27147-176] - ChIP Grade (<a href='/en-us/products/primary-antibodies/phc2-antibody-epr27147-176-chip-grade-ab314436'>ab314436</a>) at 1/1000 dilution
Lane 1:
NCCIT (human pluripotent embryonic carcinoma epithelial cell) whole cell lysate at 20 µg
Lanes 2 - 3:
Human testis tissue lysate at 20 µg
Lane 4:
Mouse testis tissue lysate at 20 µg
Lane 5:
Mouse kidney tissue lysate at 20 µg
Lane 6:
Rat testis tissue lysate at 20 µg
Lane 7:
Rat kidney tissue lysate at 20 µg
Secondary
All lanes:
Western blot - Goat Anti-Rabbit IgG H&L (HRP) (<a href='/en-us/products/secondary-antibodies/goat-rabbit-igg-h-l-hrp-ab97051'>ab97051</a>) at 1/100000 dilution
Observed band size: 31 kDa,36 kDa,40 kDa
false
- WB
Supplier Data
Western blot - Anti-PHC2 antibody [EPR27147-176] - ChIP Grade - BSA and Azide free (AB314437)
This data was developed using ab314436, the same antibody clone in a different buffer formulation. Blocking and diluting buffer and concentration : 5% NFDM/TBST The expression profile/molecular weight observed is consistent with what has been described in the literature (PMID : 16024804). In Western blot, Anti-GAPDH antibody [EPR16891] - Loading Control (ab181602) staining at 1/200000 dilution.
All lanes:
Western blot - Anti-PHC2 antibody [EPR27147-176] - ChIP Grade (<a href='/en-us/products/primary-antibodies/phc2-antibody-epr27147-176-chip-grade-ab314436'>ab314436</a>) at 1/1000 dilution
Lane 1:
SW480 (human colorectal adenocarcinoma epithelial cell) transfected with scrambled siRNA control whole cell lysate at 20 µg
Lane 2:
SW480 transfected with siRNA specifically targeti PHC2 whole cell lysate at 20 µg
Secondary
All lanes:
Western blot - Goat Anti-Rabbit IgG H&L (HRP) (<a href='/en-us/products/secondary-antibodies/goat-rabbit-igg-h-l-hrp-ab97051'>ab97051</a>) at 1/100000 dilution
Observed band size: 31 kDa,36 kDa,40 kDa
false
Exposure time: 80s
- WB
Supplier Data
Western blot - Anti-PHC2 antibody [EPR27147-176] - ChIP Grade - BSA and Azide free (AB314437)
This data was developed using ab314436, the same antibody clone in a different buffer formulation. Blocking and diluting buffer and concentration : 5% NFDM/TBST Low expression : Raji (PMID : 9121482) The expression profile/ molecular weight observed is consistent with what has been described in the literature (PMID : 16024804). In Western blot, Anti-GAPDH antibody [EPR16891] - Loading Control (ab181602) staining at 1/200000 dilution.
All lanes:
Western blot - Anti-PHC2 antibody [EPR27147-176] - ChIP Grade (<a href='/en-us/products/primary-antibodies/phc2-antibody-epr27147-176-chip-grade-ab314436'>ab314436</a>) at 1/1000 dilution
Lane 1:
SW480 (human colorectal adenocarcinoma epithelial cell) whole cell lysate at 20 µg
Lane 2:
Raji (human burkitts lymphoma b lymphocyte) whole cell lysate at 20 µg
Lane 3:
NIH/3T3 (mouse embryonic fibroblast) whole cell lysate at 20 µg
Secondary
All lanes:
Western blot - Goat Anti-Rabbit IgG H&L (HRP) (<a href='/en-us/products/secondary-antibodies/goat-rabbit-igg-h-l-hrp-ab97051'>ab97051</a>) at 1/100000 dilution
Observed band size: 31 kDa,36 kDa,40 kDa
false
Exposure time: 92s
- WB
Supplier Data
Western blot - Anti-PHC2 antibody [EPR27147-176] - ChIP Grade - BSA and Azide free (AB314437)
This data was developed using ab314436, the same antibody clone in a different buffer formulation. Blocking and diluting buffer and concentration : 5% NFDM/TBST This antibody does not react with human PHC1 and PHC3. In Western blot, Anti-6X His tag® antibody [EPR20547] - ChIP Grade (ab213204) staining at 1/5000 dilution
All lanes:
Western blot - Anti-PHC2 antibody [EPR27147-176] - ChIP Grade (<a href='/en-us/products/primary-antibodies/phc2-antibody-epr27147-176-chip-grade-ab314436'>ab314436</a>) at 1/1000 dilution
Lane 1:
His-tagged human PHC2 recombinant protein at 10 ng
Lane 2:
His-tagged human PHC1 recombinant protein at 10 ng
Lane 3:
His-tagged human PHC3 recombinant protein at 10 ng
Secondary
All lanes:
Western blot - Goat Anti-Rabbit IgG H&L (HRP) (<a href='/en-us/products/secondary-antibodies/goat-rabbit-igg-h-l-hrp-ab97051'>ab97051</a>) at 1/100000 dilution
Observed band size: 14 kDa
false
Exposure time: 48s
Related conjugates and formulations (1)
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665 Alexa Fluor® 647
Alexa Fluor® 647 Anti-PHC2 antibody [EPR27147-176] - ChIP Grade
Reactivity data
Product details
ab314437 is the carrier-free version of ab314436.
Patented technology
Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMAb® patents.
What are the advantages of a recombinant monoclonal antibody?
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.
Conjugation ready
Our carrier-free antibodies are typically supplied in a PBS-only formulation, purified and free of BSA, sodium azide and glycerol. 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.
Compatibility
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.
Properties and storage information
Form
Purification technique
Storage buffer
Shipped at conditions
Appropriate short-term storage conditions
Appropriate long-term storage conditions
Supplementary information
This supplementary information is collated from multiple sources and compiled automatically.
Biological function summary
PHC2 functions as a part of the Polycomb repressive complex 1 (PRC1). This complex is involved in modifying the structure of chromatin which helps maintain the repression of gene transcription. Through interaction with other PRC1 components PHC2 helps establish a compact chromatin state promoting histone modifications that lead to transcriptional silencing. This activity is fundamental for cellular processes like embryonic development and stem cell maintenance.
Pathways
PHC2 plays a role in the regulation of the Wnt signaling and Notch signaling pathways. These pathways are integral to controlling cell proliferation differentiation and apoptosis. Within these pathways PHC2 interacts with other proteins like BMI1 and RING1B which are also part of the PRC1 complex. These interactions ensure the precise control over gene expression needed for cellular homeostasis and proper developmental progression.
Product protocols
- Visit the General protocols
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Target data
Product promise
Please note: All products are 'FOR RESEARCH USE ONLY. NOT FOR USE IN DIAGNOSTIC OR THERAPEUTIC PROCEDURES'.
For licensing inquiries, please contact partnerships@abcam.com