- PA5-28195 - Invitrogen Antibodies OGDH antibody

Submitted references Quantitative Proteomic Approach Reveals Altered Metabolic Pathways in Response to the Inhibition of Lysine Deacetylases in A549 Cells under Normoxia and Hypoxia.

Martín-Bernabé A, Tarragó-Celada J, Cunin V, Michelland S, Cortés R, Poignant J, Boyault C, Rachidi W, Bourgoin-Voillard S, Cascante M, Seve M
International journal of molecular sciences 2021 Mar 25;22(7)

Selective knockdown of hexokinase 2 in rods leads to age-related photoreceptor degeneration and retinal metabolic remodeling.

Zhang R, Shen W, Du J, Gillies MC
Cell death & disease 2020 Oct 20;11(10):885

The Eating-Disorder Associated HDAC4(A778T) Mutation Alters Feeding Behaviors in Female Mice.

Lutter M, Khan MZ, Satio K, Davis KC, Kidder IJ, McDaniel L, Darbro BW, Pieper AA, Cui H
Biological psychiatry 2017 May 1;81(9):770-777

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Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Western Blot analysis of OGDH 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 OGDH Polyclonal Antibody (Product # PA5-28195) at a dilution of 1:1000 and a HRP-conjugated anti-rabbit IgG secondary antibody.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Knockdown of OGDH was achieved by transfecting Hep G2 with OGDH specific siRNAs (Silencer® select Product # s9834, s9835). Western blot analysis (Fig. a) was performed using membrane enriched extracts from the OGDH knockdown cells (lane 3), non-specific scrambled siRNA transfected cells (lane 2) and untransfected cells (lane 1). The blot was probed with OGDH Polyclonal Antibody (Product # PA5-28195, 1:1000 dilution) and Goat anti-Rabbit IgG (H+L) Superclonal™ Secondary Antibody, HRP conjugate (Product # A27036, 0.25µg/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 OGDH.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: Western blot analysis was performed on membrane enriched extracts (30 µg lysate) of Hep G2 (Lane 1), LNCaP (Lane 2), C2C12 (Lane 3), SW-480 (Lane 4), tissue extracts of Mouse Heart (Lane 5), Mouse Skeletal Muscle (Lane 6), Mouse Kidney (Lane 7) and Mouse Brain (Lane 8). The blot was probed with Anti-OGDH Polyclonal Antibody (Product # PA5-14358, 1:1000 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 116 kDa band corresponding to OGDH was observed across the cell lines and tissues tested.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Western Blot using OGDH Polyclonal Antibody (Product # PA5-28195). Sample (50 µg of whole cell lysate). Lane A: Mouse brain. 7.5% SDS PAGE. OGDH Polyclonal Antibody (Product # PA5-28195) diluted at 1:5,000. The HRP-conjugated anti-rabbit IgG antibody was used to detect the primary antibody.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Western Blot using OGDH Polyclonal Antibody (Product # PA5-28195). Various whole cell extracts (30 µg) were separated by 7.5% SDS-PAGE, and the membrane was blotted with OGDH Polyclonal Antibody (Product # PA5-28195) diluted at 1:1,000. The HRP-conjugated anti-rabbit IgG antibody was used to detect the primary antibody.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: OGDH antibody detects OGDH protein by western blot analysis. Various whole cell extracts (30 µg) were separated by 7.5% SDS-PAGE, and blotted with OGDH antibody OGDH Polyclonal Antibody (Product # PA5-28195) diluted by 1:1,000. The HRP-conjugated anti-rabbit IgG antibody was used to detect the primary antibody.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Western Blot analysis of OGDH 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 OGDH Polyclonal Antibody (Product # PA5-28195) at a dilution of 1:1000 and a HRP-conjugated anti-rabbit IgG secondary antibody.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Western Blot analysis of OGDH 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 OGDH Polyclonal Antibody (Product # PA5-28195) at a dilution of 1:1000 and a HRP-conjugated anti-rabbit IgG secondary antibody.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Immunofluorescence analysis of OGDH was performed using 70% confluent log phase Hep G2 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 OGDH Polyclonal Antibody (Product # PA5-28195) at 1:100 dilution 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 SlowFade® Gold Antifade Mountant with DAPI (Product # S36938). Mitochondria (Panel c: red) was stained with Mitotracker Red CMXRos (Product # M7512). Panel d represents the merged image showing mitochondrial 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: Immunocytochemistry-Immunofluorescence analysis of OGDH was performed in HeLa cells fixed in ice-cold MeOH for 5 min. Green: OGDH Polyclonal Antibody (Product # PA5-28195) diluted at 1:500. Blue: Hoechst 33342 staining. Scale bar = 10 µm.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Immunocytochemistry-Immunofluorescence analysis of OGDH was performed in HeLa cells fixed in ice-cold MeOH for 5 min. Green: OGDH Polyclonal Antibody (Product # PA5-28195) diluted at 1:500. Blue: Hoechst 33342 staining. Scale bar = 10 µm.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: OGDH Polyclonal Antibody detects OGDH protein at mitochondria by immunofluorescent analysis. Sample: HeLa cells were fixed in ice-cold MeOH for 5 min. Green: OGDH stained by OGDH Polyclonal Antibody (Product # PA5-28195) diluted at 1:200. Blue: Fluoroshield with DAPI . Scale bar= 10 µm.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Immunohistochemical analysis of paraffin-embedded Cal27 xenograft, using OGDH (Product # PA5-28195) antibody at 1:500 dilution. Antigen Retrieval: EDTA based buffer, pH 8.0, 15 min.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: OGDH Polyclonal Antibody detects OGDH protein at cytoplasm by immunohistochemical analysis. Sample: Paraffin-embedded mouse heart. OGDH stained by OGDH Polyclonal Antibody (Product # PA5-28195) diluted at 1:500. Antigen Retrieval: Citrate buffer, pH 6.0, 15 min.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: OGDH Polyclonal Antibody detects OGDH protein at cytoplasm by immunohistochemical analysis. Sample: Paraffin-embedded mouse muscle. OGDH stained by OGDH Polyclonal Antibody (Product # PA5-28195) diluted at 1:500. Antigen Retrieval: Citrate buffer, pH 6.0, 15 min.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: OGDH Polyclonal Antibody detects OGDH protein at mitochondria on mouse muscle by immunohistochemical analysis. Sample: Paraffin-embedded mouse muscle. OGDH Polyclonal Antibody (Product # PA5-28195) dilution: 1:500. Antigen Retrieval: EDTA based buffer, pH 8.0, 15 min.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Fig. 6 Knocking down HK2 led to upregulation of enzymes in the TCA cycle. A - D Immunostaining for enzymes in the TCA cycle including pyruvate dehydrogenase E1alpha (PDHE1alpha, A and B ) and oxoglutarate dehydrogenase (OGDH, C and D ) in retinal sections from HKKO mice and age-matched WT controls. Loss of HK2 in rods led to increased expression of PDHE1alpha ( A , B ) and OGDH ( C , D ) in the photoreceptor inner segments (PIS). The panel of images in ( B ) and ( D ) are higher-power images of areas in ( A ) and ( C ) respectively. Scale bar: 50 mum in ( A - D ). E , F Western blots indicated that the expression of PDHE1alpha ( C ) and OGDH ( D ) was significantly increased in HKKO mice compared with age-matched WT controls. * p < 0.05 and ** p < 0.01, n = 4/group.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Figure 4 Effect of KDAC inhibition on enzyme activities in A549 cells under normoxia and hypoxia. ( A , B ) The ATP- dependent 6-phosphofructokinase (PFK1) ( A ) and lactate dehydrogenase (LDH) ( B ) enzymatic activities were measured after 24 h of incubation, and activities were normalized to intracellular protein content in each condition. A549 cells were treated with 1 muM of TSA, 20 mM of NAM, and both 1 muM TSA and 20 mM NAM for 24 h of incubation under normoxia and hypoxia. Cells incubated in medium without KDACIs served as control. Bars represent the means +- standard error of the mean of three independent experiments. The asterisks above bars indicate statistically significant differences compared to normoxic control cells. The asterisks above curly brackets indicate statistically significant differences between hypoxic and normoxic treatments and between hypoxic treatments and hypoxic control cells. Statistical significance was assessed by a two-tailed Student''s t -test. *, p

PA5-28195

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