Anti-DIAPH1 antibody [EPR7948] - BSA and Azide free (ab248325)

Rabbit Recombinant Monoclonal DIAPH1 antibody. Carrier free. Suitable for IHC-P, ICC/IF, WB, Flow Cyt (Intra) and reacts with Mouse, Human, Rat samples.

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Images

Western blot - Anti-DIAPH1 antibody [EPR7948] - BSA and Azide free (AB248325), expandable thumbnail

Key facts

Isotype: IgG
Host species: Rabbit
Storage buffer: pH: 7.2 - 7.4
Constituents: 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

	IHC-P	ICC/IF	WB	Flow Cyt (Intra)
Human	Tested	Tested	Tested	Tested
Mouse	Tested	Expected	Tested	Expected
Rat	Tested	Expected	Tested	Expected

Tested
Tested

Species	Dilution info	Notes
Species Mouse	Dilution info -	Notes Perform heat-mediated antigen retrieval before commencing with IHC staining protocol.
Species Human	Dilution info -	Notes Perform heat-mediated antigen retrieval before commencing with IHC staining protocol.
Species Rat	Dilution info -	Notes Perform heat-mediated antigen retrieval before commencing with IHC staining protocol.

Tested
Tested

Species	Dilution info	Notes
Species Human	Dilution info -	Notes -

Expected
Expected

Species	Dilution info	Notes
Species Mouse, Rat	Dilution info Use at an assay dependent concentration.	Notes -

Tested
Tested

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

Tested
Tested

Species	Dilution info	Notes
Species Human	Dilution info -	Notes -

Expected
Expected

Species	Dilution info	Notes
Species Mouse, Rat	Dilution info Use at an assay dependent concentration.	Notes -

Associated Products

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Target data

See full target information DIAPH1

Function

Actin nucleation and elongation factor required for the assembly of F-actin structures, such as actin cables and stress fibers (By similarity). Binds to the barbed end of the actin filament and slows down actin polymerization and depolymerization (By similarity). Required for cytokinesis, and transcriptional activation of the serum response factor (By similarity). DFR proteins couple Rho and Src tyrosine kinase during signaling and the regulation of actin dynamics (By similarity). Functions as a scaffold protein for MAPRE1 and APC to stabilize microtubules and promote cell migration (By similarity). Has neurite outgrowth promoting activity. Acts in a Rho-dependent manner to recruit PFY1 to the membrane (By similarity). In hear cells, it may play a role in the regulation of actin polymerization in hair cells (PubMed:20937854, PubMed:21834987, PubMed:26912466). The MEMO1-RHOA-DIAPH1 signaling pathway plays an important role in ERBB2-dependent stabilization of microtubules at the cell cortex (PubMed:20937854, PubMed:21834987). It controls the localization of APC and CLASP2 to the cell membrane, via the regulation of GSK3B activity (PubMed:20937854, PubMed:21834987). In turn, membrane-bound APC allows the localization of the MACF1 to the cell membrane, which is required for microtubule capture and stabilization (PubMed:20937854, PubMed:21834987). Plays a role in the regulation of cell morphology and cytoskeletal organization. Required in the control of cell shape (PubMed:20937854, PubMed:21834987). Plays a role in brain development (PubMed:24781755). Also acts as an actin nucleation and elongation factor in the nucleus by promoting nuclear actin polymerization inside the nucleus to drive serum-dependent SRF-MRTFA activity (By similarity).

Alternative names

DIAP1, DIAPH1, Protein diaphanous homolog 1, Diaphanous-related formin-1, DRF1

Recommended products

Rabbit Recombinant Monoclonal DIAPH1 antibody. Carrier free. Suitable for IHC-P, ICC/IF, WB, Flow Cyt (Intra) and reacts with Mouse, Human, Rat samples.

Key facts

Isotype: IgG
Host species: Rabbit
Storage buffer: pH: 7.2 - 7.4
Constituents: PBS
Form: Liquid
Clonality: Monoclonal
Immunogen: The exact immunogen used to generate this antibody is proprietary information.
Carrier free: Yes
Clone number: EPR7948
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
Storage information: Do Not Freeze

Notes

ab248325 is the carrier-free version of Anti-DIAPH1 antibody [EPR7948] ab129167.

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

DIAPH1 also known as Diaphanous-related formin 1 operates mainly in actin polymerization and is critical for filament formation. It weighs approximately 140 kDa. DIAPH1 expresses in a variety of tissues including heart liver and kidney making it a target of interest in multiple biological contexts. The protein plays a role in maintaining proper cytoskeletal dynamics by regulating the assembly of actin filaments.

Biological function summary

DIAPH1 functions as part of the formin family which are important in cytoskeletal organization. It interacts with other proteins to initiate and elongate actin filaments often acting within larger protein complexes that control cellular morphology and movement. DIAPH1 contributes to muscle contraction cell division and cell migration highlighting its importance in cellular structure and dynamics.

Pathways

DIAPH1 plays a significant role in the Rho GTPase signaling pathway. This pathway connects to cytoskeleton remodeling impacting cell shape and movement. DIAPH1 associates with proteins like RhoA which regulates its activity. Additionally DIAPH1 influences the Wnt signaling pathway known for its involvement in cell proliferation and differentiation.

Associated diseases and disorders

DIAPH1 is connected to a form of sensorineural hearing loss and leukemia. Mutations in DIAPH1 can lead to autosomal dominant deafness as it affects the proper maintenance of actin structures in hair cells within the inner ear. The protein also interacts with genes linked to cancer development including those coding for p53 influencing tumorigenesis and cell cycle regulation.

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:
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8 product images

Western blot - Anti-DIAPH1 antibody [EPR7948] - BSA and Azide free (ab248325)
All lanes: Western blot - Anti-DIAPH1 antibody [EPR7948] (Anti-DIAPH1 antibody [EPR7948] ab129167) at 1/10000 dilution
Lane 1: PC-12 (Rat adrenal gland pheochromocytoma ) whole cell lysate at 20 µg
Lane 2: MCF7 (Human breast adenocarcinoma epithelial cell) whole cell lysate at 20 µg
Secondary
All lanes: Goat Anti-Rabbit IgG (HRP) with minimal cross-reactivity with human IgG at 1/2000 dilution
Predicted band size: 141 kDa
Western blot - Anti-DIAPH1 antibody [EPR7948] - BSA and Azide free (ab248325)
All lanes: Western blot - Anti-DIAPH1 antibody [EPR7948] (Anti-DIAPH1 antibody [EPR7948] ab129167) at 1/1000 dilution
Lane 1: HEK-293 (Human embryonic kidney epithelial cell) whole cell lysate at 20 µg
Lane 2: Human brain lysate at 20 µg
Lane 3: RAW 264.7 (Mouse Abelson murine leukemia virus-induced tumor macrophage) whole cell lysate at 20 µg
Secondary
All lanes: Goat Anti-Rabbit IgG (HRP) with minimal cross-reactivity with human IgG at 1/2000 dilution
Predicted band size: 141 kDa
Western blot - Anti-DIAPH1 antibody [EPR7948] - BSA and Azide free (ab248325)
This data was developed using the same antibody clone in a different buffer formulation (Anti-DIAPH1 antibody [EPR7948] ab129167).
Lanes 1- 2: Merged signal (red and green). Green - Anti-DIAPH1 antibody [EPR7948] ab129167 observed at 150 kDa. Red - Anti-alpha Tubulin antibody [DM1A] - Loading Control (Anti-alpha Tubulin antibody [DM1A] - Loading Control ab7291) observed at 50 kDa.
Anti-DIAPH1 antibody [EPR7948] ab129167 was shown to react with DIAPH1 in wild-type HEK-293T cells in western blot. Loss of signal was observed when knockout cell line Human DIAPH1 knockout HEK-293T cell line ab266120 (knockout cell lysate Human DIAPH1 knockout HEK-293T cell lysate ab257411) was used. Wild-type HEK-293T and DIAPH1 knockout HEK-293T cell lysates were subjected to SDS-PAGE. Membrane was blocked for 1 hour at room temperature in 0.1% TBST with 3% non-fat dried milk. Anti-DIAPH1 antibody [EPR7948] ab129167 and Anti-alpha Tubulin antibody [DM1A] - Loading Control (Anti-alpha Tubulin antibody [DM1A] - Loading Control ab7291) overnight at 4°C at a 1 in 1000 dilution and a 1 in 20000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye^®800CW) preadsorbed (Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed ab216773) and Goat anti-Mouse IgG H&L (IRDye^®680RD) preadsorbed (Goat anti-Mouse IgG H&L (IRDye® 680RD) preadsorbed ab216776) secondary antibodies at 1 in 20000 dilution for 1 hour at room temperature before imaging.
All lanes: Western blot - Anti-DIAPH1 antibody [EPR7948] (Anti-DIAPH1 antibody [EPR7948] ab129167) at 1/1000 dilution
Lane 1: Wild-type HEK-293T cell lysate at 20 µg
Lane 2: DIAPH1 knockout HEK-293T cell lysate at 20 µg
Lane 2: Western blot - Human DIAPH1 knockout HCT116 cell line (Human DIAPH1 knockout HCT116 cell line ab273727)
Performed under reducing conditions.
Predicted band size: 141 kDa
Observed band size: 150 kDa
Immunocytochemistry/ Immunofluorescence - Anti-DIAPH1 antibody [EPR7948] - BSA and Azide free (ab248325)
This data was developed using Anti-DIAPH1 antibody [EPR7948] ab129167, the same antibody clone in a different buffer formulation.
Immunocytochemistry analysis of HeLa (Human cervix adenocarcinoma epithelial cell) cells labeling DIAPH1 with Purified Anti-DIAPH1 antibody [EPR7948] ab129167 at 1/50 dilution (10 µg/mL). Cells were fixed in 4% Paraformaldehyde and permeabilized with 0.1% tritonX-100. Cells were counterstained with Alexa Fluor® 594 Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker ab195889 Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor® 594) 1/200 dilution (2.5 µg/mL). Goat anti rabbit IgG (Alexa Fluor® 488, Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) ab150077) was used as the secondary antibody at 1/1000 dilution (2 µg/mL). DAPI (blue) was used as nuclear counterstain. PBS instead of the primary antibody was used as the secondary antibody only control.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-DIAPH1 antibody [EPR7948] - BSA and Azide free (ab248325)
This data was developed using Anti-DIAPH1 antibody [EPR7948] ab129167, the same antibody clone in a different buffer formulation.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human kidney tissue sections labeling DIAPH1 with Purified Anti-DIAPH1 antibody [EPR7948] ab129167 at 1/100 dilution (5.96 µg/mL). Heat mediated antigen retrieval using Bond™ Epitope Retrieval Solution 2 (pH 9.0). Rabbit specific IHC polymer detection kit HRP/DAB (Rabbit specific IHC polymer detection kit HRP/DAB ab209101) was used as the secondary antibody. Negative control: PBS instead of the primary antibody. Hematoxylin was used as a counterstain.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-DIAPH1 antibody [EPR7948] - BSA and Azide free (ab248325)
This data was developed using Anti-DIAPH1 antibody [EPR7948] ab129167, the same antibody clone in a different buffer formulation.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of mouse kidney tissue sections labeling DIAPH1 with Purified Anti-DIAPH1 antibody [EPR7948] ab129167 at 1/100 dilution (5.96 µg/mL). Heat mediated antigen retrieval using Bond™ Epitope Retrieval Solution 2 (pH 9.0). Rabbit specific IHC polymer detection kit HRP/DAB (Rabbit specific IHC polymer detection kit HRP/DAB ab209101) was used as the secondary antibody. Negative control: PBS instead of the primary antibody. Hematoxylin was used as a counterstain.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-DIAPH1 antibody [EPR7948] - BSA and Azide free (ab248325)
This data was developed using Anti-DIAPH1 antibody [EPR7948] ab129167, the same antibody clone in a different buffer formulation.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of rat kidney tissue sections labeling DIAPH1 with Purified Anti-DIAPH1 antibody [EPR7948] ab129167 at 1/100 dilution (5.96 µg/mL). Heat mediated antigen retrieval using Bond™ Epitope Retrieval Solution 2 (pH 9.0). Rabbit specific IHC polymer detection kit HRP/DAB (Rabbit specific IHC polymer detection kit HRP/DAB ab209101) was used as the secondary antibody. Negative control: PBS instead of the primary antibody. Hematoxylin was used as a counterstain.
Flow Cytometry (Intracellular) - Anti-DIAPH1 antibody [EPR7948] - BSA and Azide free (ab248325)
This data was developed using Anti-DIAPH1 antibody [EPR7948] ab129167, the same antibody clone in a different buffer formulation.
Intracellular Flow Cytometry analysis of HeLa (human epithelial cell line from cervix adenocarcinoma) cells labeling DIAPH1 with purified Anti-DIAPH1 antibody [EPR7948] ab129167 at 1/50 dilution. Cells were fixed with 4% paraformaldehyde and permeabilised with 90% methanol. Goat Anti-Rabbit IgG H&L (Alexa Fluor^® 488) (Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) ab150077) at 1/2000 was used as the secondary antibody (red). Rabbit monoclonal IgG (Rabbit IgG, monoclonal [EPR25A] - Isotype Control ab172730) was used as the isotype control (black). Cells without incubation with primary and secondary antibodies were used as the unlabeled control (blue).