Rabbit Recombinant Monoclonal DIAPH1 antibody. Suitable for IHC-P, ICC/IF, WB, Flow Cyt (Intra) and reacts with Mouse, Rat, Human samples. Cited in 15 publications.
pH: 7.2 - 7.4
Preservative: 0.01% Sodium azide
Constituents: 59% PBS, 40% Glycerol (glycerin, glycerine), 0.05% BSA
IHC-P | ICC/IF | WB | Flow Cyt (Intra) | |
---|---|---|---|---|
Human | Tested | Tested | Tested | Tested |
Mouse | Tested | Expected | Expected | Expected |
Rat | Tested | Expected | Tested | Expected |
Species | Dilution info | Notes |
---|---|---|
Species Mouse | Dilution info 1/50 - 1/100 | Notes Perform heat-mediated antigen retrieval before commencing with IHC staining protocol. |
Species Rat | Dilution info 1/50 - 1/100 | Notes Perform heat-mediated antigen retrieval before commencing with IHC staining protocol. |
Species Human | Dilution info 1/50 - 1/100 | Notes Perform heat-mediated antigen retrieval before commencing with IHC staining protocol. |
Species | Dilution info | Notes |
---|---|---|
Species Human | Dilution info 1/50 | Notes For unpurified use at 1/100 - 1/250. |
Species | Dilution info | Notes |
---|---|---|
Species Mouse, Rat | Dilution info Use at an assay dependent concentration. | Notes - |
Species | Dilution info | Notes |
---|---|---|
Species Rat | Dilution info 1/1000 - 1/10000 | Notes - |
Species Human | Dilution info 1/1000 - 1/10000 | Notes - |
Species | Dilution info | Notes |
---|---|---|
Species Mouse | Dilution info Use at an assay dependent concentration. | Notes - |
Species | Dilution info | Notes |
---|---|---|
Species Human | Dilution info 1/50 | Notes - |
Species | Dilution info | Notes |
---|---|---|
Species Mouse, Rat | Dilution info Use at an assay dependent concentration. | Notes - |
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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).
DIAP1, DIAPH1, Protein diaphanous homolog 1, Diaphanous-related formin-1, DRF1
Rabbit Recombinant Monoclonal DIAPH1 antibody. Suitable for IHC-P, ICC/IF, WB, Flow Cyt (Intra) and reacts with Mouse, Rat, Human samples. Cited in 15 publications.
pH: 7.2 - 7.4
Preservative: 0.01% Sodium azide
Constituents: 59% PBS, 40% Glycerol (glycerin, glycerine), 0.05% BSA
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:
For more information, read more on recombinant antibodies.
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.
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.
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.
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.
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We have not tested this specific species and application combination in-house, but expect it will work. It is covered by our product promise.
This species and application combination has not been tested, but we predict it will work based on strong homology. However, this combination is not covered by our product promise.
We do not recommend this combination. It is not covered by our product promise.
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Terms & Conditions.
ab129167 was shown to react with DIAPH1 in wild-type HCT 116 cells in western blot with loss of signal observed in DIAPH1 knockout cell line Human DIAPH1 knockout HCT116 cell line ab273727 (DIAPH1 knockout cell lysate Human DIAPH1 knockout HCT116 cell lysate ab275252). Wild-type and DIAPH1 knockout HCT 116 cell lysates were subjected to SDS-PAGE. Membranes were blocked in 3% milk in TBS-T (0.1% Tween®) before incubation with ab129167 and Anti-GAPDH antibody [6C5] - Loading Control ab8245 (Mouse anti-GAPDH antibody [6C5]) overnight at 4°C at a 1 in 1000 dilution and a 1 in 20000 dilution respectively. Blots were incubated with Goat anti-Rabbit IgG H&L (IRDye® 800CW) preabsorbed (Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed ab216773) and Goat anti-Mouse IgG H&L (IRDye® 680RD) preabsorbed (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] (ab129167) at 1/1000 dilution
Lane 1: Wild-type HCT116 cell lysate at 20 µg
Lane 2: DIAPH1 knockout HCT116 cell lysate at 20 µg
Lane 2: Western blot - Human DIAPH1 knockout HEK-293T cell line (Human DIAPH1 knockout HEK-293T cell line ab266120)
Performed under reducing conditions.
Predicted band size: 141 kDa
Observed band size: 155 kDa
All lanes: Western blot - 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
All lanes: Goat Anti-Rabbit IgG (HRP) with minimal cross-reactivity with human IgG at 1/2000 dilution
Predicted band size: 141 kDa
All lanes: Western blot - 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
All lanes: Goat Anti-Rabbit IgG (HRP) with minimal cross-reactivity with human IgG at 1/2000 dilution
Predicted band size: 141 kDa
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. 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] (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 analysis of HeLa (Human cervix adenocarcinoma epithelial cell) cells labeling DIAPH1 with Purified 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) analysis of human kidney tissue sections labeling DIAPH1 with Purified 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) analysis of mouse kidney tissue sections labeling DIAPH1 with Purified 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) analysis of rat kidney tissue sections labeling DIAPH1 with Purified 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.
Intracellular Flow Cytometry analysis of HeLa (human epithelial cell line from cervix adenocarcinoma) cells labeling DIAPH1 with purified 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).
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