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antibody from Invitrogen Antibodies
Targeting: ARHGEF2
GEF-H1, GEFH1, KIAA0651, Lfc, LFP40, P40
Western blot
ImmunoprecipitationAntibody data
- Antibody Data
- Antigen structure
- References [0]
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- Validations
- Western blot [10]
- Immunocytochemistry [2]
- Immunohistochemistry [5]
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- Product number
- PA5-32213 - Provider product page
- Provider
- Invitrogen Antibodies
- Product name
- GEF-H1 Polyclonal Antibody
- Antibody type
- Polyclonal
- Antigen
- Recombinant protein fragment
- Description
- Recommended positive controls: 293T, A431, BCL-1, C2C12, C8D30, GL261, HeLa, HepG2, NIH-3T3, Neuro 2A, Raw264.7, hMSC-3A6, PC-12, Rat2, mouse fibroblast. Predicted reactivity: Mouse (89%), Rat (89%), Dog (92%), Pig (95%), Bovine (94%). Store product as a concentrated solution. Centrifuge briefly prior to opening the vial.
- Reactivity
- Human, Mouse, Rat
- Host
- Rabbit
- Isotype
- IgG
- Vial size
- 100 µL
- Concentration
- 0.51 mg/mL
- Storage
- Store at 4°C short term. For long term storage, store at -20°C, avoiding freeze/thaw cycles.
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Supportive validation
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Western blot analysis of GEF-H1 using 10 µg of A) hMSC-3A6 and B) siRNA of ARHGEF2/GEF-H1 lysate. Samples were loaded onto a 8% SDS-PAGE gel and probed with a GEF-H1 polyclonal antibody (Product # PA5-32213) at a dilution of 1:1000.
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Western Blot analysis of GEF-H1 was performed by separating 30 µg of various whole cell extracts by 7.5% SDS-PAGE. Proteins were transferred to a membrane and probed with a GEF-H1 Polyclonal Antibody (Product # PA5-32213) at a dilution of 1:2000 and a HRP-conjugated anti-rabbit IgG secondary antibody.
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Western Blot using GEF-H1 Polyclonal Antibody (Product # PA5-32213). Sample (10 µg of whole cell lysate). Lane A: hMSC-3A6. Lane B: siRNA of ARHGEF2/GEF-H1. 8% SDS PAGE. GEF-H1 Polyclonal Antibody (Product # PA5-32213) diluted at 1:1,000. The HRP-conjugated anti-rabbit IgG antibody was used to detect the primary antibody.
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- GEF-H1 Polyclonal Antibody detects GEF-H1 protein by western blot analysis. A. 30 µg Raw264.7 whole cell lysate/extract. B. 30 µg C2C12 whole cell lysate/extract.5% SDS-PAGE. GEF-H1 Polyclonal Antibody (Product # PA5-32213) dilution: 1:1,000. The HRP-conjugated anti-rabbit IgG antibody was used to detect the primary antibody.
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Western Blot analysis of GEF-H1 was performed by separating 30 µg of various whole cell extracts by 7.5% SDS-PAGE. Proteins were transferred to a membrane and probed with a GEF-H1 Polyclonal Antibody (Product # PA5-32213) at a dilution of 1:500 and a HRP-conjugated anti-rabbit IgG secondary antibody.
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Western Blot using GEF-H1 Polyclonal Antibody (Product # PA5-32213). Sample (30 µg of whole cell lysate). Lane A: 293T. Lane B: A431. Lane C: HeLa. 5% SDS PAGE. GEF-H1 Polyclonal Antibody (Product # PA5-32213) diluted at 1:2,000. The HRP-conjugated anti-rabbit IgG antibody was used to detect the primary antibody.
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Western Blot analysis of GEF-H1 was performed by separating 30 µg of various whole cell lysates by 5% SDS-PAGE. Proteins were transferred to a membrane and probed with a GEF-H1 Polyclonal Antibody (Product # PA5-32213) at a dilution of 1:2000. The HRP-conjugated anti-rabbit IgG antibody was used to detect the primary antibody. A. 293T, B. A431 , C. HeLa , D. HepG2.
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Western Blot analysis of GEF-H1 was performed by separating 30 µg of various whole cell lysates by 5% SDS-PAGE. Proteins were transferred to a membrane and probed with a GEF-H1 Polyclonal Antibody (Product # PA5-32213) at a dilution of 1:1000. The HRP-conjugated anti-rabbit IgG antibody was used to detect the primary antibody. A. Neuro2A, B. GL261 , C. C8D30 , D. NIH-3T3 , E. BCL-1 , F. Raw 264.7 , G. C2Cl2.
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Knockdown of GEFH1 was achieved by transfecting HeLa cells with GEFH1 specific siRNAs (Silencer® select Product # s17547, s17545). Western blot analysis (Fig. a) was performed using whole cell extracts from the GEFH1 knockdown cells (lane 3), non-specific scrambled siRNA transfected cells (lane 2) and untransfected cells (lane 1). The blots were probed with GEFH1 Polyclonal Antibody (Product # PA5-32213, 1:1000 dilution) and Goat anti-Rabbit IgG (H+L) Superclonal™ Secondary Antibody, HRP conjugate (Product # A27036, 0.25 µg/mL, 1:4000 dilution). Densitometric analysis of this western blot is shown in histogram (Fig. b). Decrease in signal upon siRNA mediated knock down confirms that antibody is specific to GEFH1.
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Western blot analysis was performed on whole cell extracts (30 µg lysate) of K-562 (Lane 1), SH-SY5Y (Lane 2), RAW 264.7 (Lane 3), C2C12 (Lane 4), HeLa (Lane 5) and NIH/3T3(Lane 6). The blot was probed with Anti-GEF-H1 Polyclonal Antibody (Product # PA5-32213, 1:2000 dilution) and detected by chemiluminescence using Goat anti Rabbit IgG (H+L) Superclonal™ Secondary Antibody, HRP conjugate (Product # A27036, 0.25 µg/mL, 1:4000 dilution). A 120 kDa band corresponding to GEF-H1 was observed across all cell lines tested.
Supportive validation
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Immunocytochemistry-Immunofluorescence analysis of GEF-H1 was performed in HeLa cells fixed in ice-cold MeOH for 5 min. Green: GEF-H1 Polyclonal Antibody (Product # PA5-32213) diluted at 1:500. Blue: Hoechst 33342 staining. Scale bar = 10 µm.
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Immunofluorescence analysis of GEF-H1 was performed using 70% confluent log phase K-562 cells. The cells were fixed with 4% paraformaldehyde for 10 minutes, permeabilized with 0.1% Triton™ X-100 for 15 minutes, and blocked with 1% BSA for 1 hour at room temperature. The cells were labeled with GEF-H1 Rabbit Polyclonal Antibody (Product # PA5-32213) at 5 microgram/mL in 0.1% BSA, incubated at 4 degree Celsius overnight and then labeled with Goat anti-Rabbit IgG (H+L) Superclonal™ Secondary Antibody, Alexa Fluor® 488 conjugate (Product # A27034) at a dilution of 1:2000 for 45 minutes at room temperature (Panel a: green). Nuclei (Panel b: blue) were stained with ProLong™ Diamond Antifade Mountant with DAPI (Product # P36962). F-actin (Panel c: red) was stained with Rhodamine Phalloidin (Product # R415). Panel d represents the merged image showing cytoplasmic localization. Panel e represents control cells with no primary antibody to assess background. The images were captured at 60X magnification.
Supportive validation
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Immunohistochemistry (Paraffin) analysis of GEF-H1 was performed in paraffin-embedded mouse brain tissue using GEF-H1 Polyclonal Antibody (Product # PA5-32213) at a dilution of 1:1000. Antigen Retrieval: Citrate buffer, pH 6.0, 15 min.
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Immunohistochemistry (Frozen) analysis of GEF-H1 was performed in frozen-sectioned mouse mouse cerebellum tissue using GEF-H1 Polyclonal Antibody (Product # PA5-32213) at a dilution of 1:250 (Green). Red: NF-H, stained by NF-H antibody diluted at 1:500. Blue: Fluoroshield with DAPI. Antigen Retrieval: Citrate buffer, pH 6.0, 10 min.
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- GEF-H1 Polyclonal Antibody detects GEF-H1 protein at cytosol on mouse testis by immunohistochemical analysis. Sample: Paraffin-embedded mouse testis. GEF-H1 Polyclonal Antibody (Product # PA5-32213) dilution: 1:500. Antigen Retrieval: EDTA based buffer, pH 8.0, 15 min.
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- GEF-H1 Polyclonal Antibody detects GEF-H1 protein at cytosol on rat hind brain by immunohistochemical analysis. Sample: Paraffin-embedded rat hind brain. GEF-H1 Polyclonal Antibody (Product # PA5-32213) dilution: 1:500. Antigen Retrieval: EDTA based buffer, pH 8.0, 15 min.
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Immunohistochemical analysis of paraffin-embedded human gastric cancer, using GEF-H1 (Product # PA5-32213) antibody at 1:500 dilution. Antigen Retrieval: EDTA based buffer, pH 8.0, 15 min.
