- PA1-16777 - Invitrogen Antibodies GAPDHS antibody

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Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: Western blot analysis was performed on whole cell extracts (30µg lysate) of A-431 (Lane 1), COS-7 (Lane 2), MDCK (Lane 3), C2C12 (Lane 4), PC-3 (Lane 5), PC-12 (Lane 6), Neuro-2a (Lane 7), tissue extracts (30µg lysate) of Mouse Liver (Lane 8), Rat Stomach (Lane 9), Mouse Pancreas (Lane 10) and Rat Heart (Lane 11). The blots were probed with Anti-GAPDH Rabbit Polyclonal Antibody (Product # PA1-16777, 0.5µg/mL) and detected by chemiluminescence using Goat anti-Rabbit IgG (H+L) Superclonal™ Secondary Antibody, HRP conjugate (Product # A27036, 0.25µg/mL, 1:4000 dilution). A 37 kDa band corresponding to GAPDH was observed across the cell lines and tissues tested. Known quantity of protein samples were electrophoresed using Novex® NuPAGE® 4-12 % Bis-Tris gel (Product # NP0321BOX), XCell SureLock™ Electrophoresis System (Product # EI0002) and Novex® Sharp Pre-Stained Protein Standard (Product # LC5800). Resolved proteins were then transferred onto a nitrocellulose membrane with iBlot® 2 Dry Blotting System (Product # IB21001). The membrane was probed with the relevant primary and secondary Antibody following blocking with 5 % skimmed milk. Chemiluminescent detection was performed using Pierce™ ECL Western Blotting Substrate (Product # 32106).

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: Western blot analysis of GAPDH in 15 µg whole cell lysate from HeLa, 293T, Jurkat, mouse TCMK-1, and mouse NIH3T3 cells. Samples were incubated in GAPDH polyclonal antibody (Product # PA1-16777 using a dilution of 0.1 µg/mL. Detection: Chemiluminescence with an exposure time of 30 seconds.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: Western blot analysis of GAPDH in whole cell lysate from mouse NIH3T3 (5, 15 and 50 µg) and human HeLa (H; 50 µg) cells. Samples were incubated in GAPDH polyclonal antibody (Product # PA1-16777 using a dilution of 0.04 µg/mL. Detection: Chemiluminescence with an exposure time of 30 seconds.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: Western blot analysis of GAPDH in 1.0 mg/mL HeLa lysate. Samples were incubated in GAPDH polyclonal antibody (Product # PA1-16777). This experiment was performed under reducing conditions using the 12-230 kDa separation system. Note: band observed higher than predicted molecular weight of 36 kDa.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: Knockdown of GAPDH was achieved by transfecting HeLa cells with GAPDH specific validated siRNAs (Silencer® select Product # s13960 ). Western blot analysis (Fig. a) was performed using whole cell extracts from the GAPDH knockdown cells (lane 3), non-specific scrambled siRNA transfected cells (lane 2) and untransfected cells (lane 1). The blots were probed with GAPDH Polyclonal Antibody (Product # PA1-16777, 0.5 µg/mL) 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 GAPDH.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: Immunocytochemistry analysis of GAPDH in HeLa cells. Samples were incubated in GAPDH polyclonal antibody (Product # PA1-16777) followed by DyLight Fluor 488.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: Immunofluorescence analysis of GAPDH was performed using 70% confluent log phase HeLa cells. The cells were fixed with 4% paraformaldehyde for 10 minutes, permeabilized with 0.1% Triton™ X-100 for 10 minutes, and blocked with 1% BSA for 1 hour at room temperature. The cells were labeled with GAPDH Polyclonal Antibody (Product # PA1-16777) at 5 µg/mL in 0.1% BSA 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). F-actin (Panel c: red) was stained with Rhodamine Phalloidin (Product # R415, 1:300) at a dilution of 1:300 for 45 minutes at room temperature. Panel d represents the merged image showing cytoplasmic localization. Panel e shows no primary antibody control. The images were captured at 60X magnification.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: Immunofluorescence analysis of GAPDH was performed using 70% confluent log phase NIH/3T3 cells. The cells were fixed with 4% paraformaldehyde for 10 minutes, permeabilized with 0.1% Triton™ X-100 for 10 minutes, and blocked with 1% BSA for 1 hour at room temperature. The cells were labeled with GAPDH Rabbit Polyclonal Antibody (Product # PA1-16777) at 2µg/mL in 0.1% BSA and incubated for 3 hours at room temperature 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). F-actin (Panel c: red) was stained with Rhodamine Phalloidin (Product # R415, 1:300). Panel d represents the merged image showing cytoplasmic localization. Panel e shows the no primary antibody control. The images were captured at 60X magnification.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: Immunofluorescence analysis of GAPDH was performed using 70% confluent log phase U2OS cells. The cells were fixed with 4% paraformaldehyde for 10 minutes, permeabilized with 0.1% Triton™ X-100 for 10 minutes, and blocked with 1% BSA for 1 hour at room temperature. The cells were labeled with GAPDH Rabbit Polyclonal Antibody (Product # PA1-16777) at 2µg/mL in 0.1% BSA and incubated for 3 hours at room temperature 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). F-actin (Panel c: red) was stained with Rhodamine Phalloidin (Product # R415, 1:300). Panel d represents the merged image showing cytoplasmic localization. Panel e shows the no primary antibody control. The images were captured at 60X magnification.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: Immunohistochemical analysis of GAPDH in formalin-fixed paraffin-embedded section of human colon carcinoma. Samples were incubated in GAPDH polyclonal antibody (Product # PA1-16777). Detection: DAB.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: Immunohistochemical analysis of GAPDH in formalin-fixed paraffin-embedded section of mouse plasmacytoma. Samples were incubated in GAPDH polyclonal antibody (Product # PA1-16777). Detection: DAB.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: Immunohistochemical analysis of GAPDH in formalin fixed paraffin embedded section of mouse squamous cell carcinoma. Samples were incubated in GAPDH polyclonal antibody (Product # PA1-16777) using a dilution of 1:200.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: Immunohistochemical analysis of GAPDH in formalin fixed paraffin embedded section of human lung carcinoma. Samples were incubated in GAPDH polyclonal antibody (Product # PA1-16777) using a dilution of 1:200.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: NULL

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: NULL

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: Figure 3 DNA uptake by DCs analyzed by flow cyometry and confocal microscopy (A) Intracellular flow cytometric detection of tumor DNA (EdU-labeled) within FLT-3L BMDCs after a 2 h incubation with tumor debris generated by heat shock (HS). Data shown as mean +- SD, with ****p < 0.0001 determined by Student's t test. (B) Stacked confocal microscopy images displaying EdU-labeled exogenous DNA (red), nuclei (green) and GAPDH (blue) in FLT-3L BMDCs treated with tumor debris generated by heat shock (HS). Scale bar: 25 mum.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: Figure 5 Wk-MTA1 plays an essential role in WHx-induced wk-NF-kappaB activation P65 nuclear translocation was assessed in WCH17 ( A ) and KD/wk-MTA1 WCH17 (designated as KD) ( B ) cells. The expression levels of wk-p65, WHx, and wk-MTA1 were determined using Western blotting. The relative ratio between nuclear (Nu) and cytoplasmic (Cy) wk-p65 was calculated using a densitometric analysis of the corresponding bands in Western blotting. Moreover, GAPDH and hnRNP served as markers for cytoplasmic and nuclear loading controls, respectively. CO: cells cotransfected with WHx and wk-MTA1-expressing plasmids ( * P < .05; ** P < .01).

PA1-16777

Antibody data