- PA5-32228 - Invitrogen Antibodies SHMT2 antibody

Submitted references Shmt2: A Stat3 Signaling New Player in Prostate Cancer Energy Metabolism.

Marrocco I, Altieri F, Rubini E, Paglia G, Chichiarelli S, Giamogante F, Macone A, Perugia G, Magliocca FM, Gurtner A, Maras B, Ragno R, Patsilinakos A, Manganaro R, Eufemi M
Cells 2019 Sep 6;8(9)

Mitochondrial dysfunction remodels one-carbon metabolism in human cells.

Bao XR, Ong SE, Goldberger O, Peng J, Sharma R, Thompson DA, Vafai SB, Cox AG, Marutani E, Ichinose F, Goessling W, Regev A, Carr SA, Clish CB, Mootha VK
eLife 2016 Jun 16;5

No comments: Submit comment

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Western Blot using SHMT2 Polyclonal Antibody (Product # PA5-32228). Sample (whole cell lysate). Lane A: 293T 10 µg. Lane B: HeLa 10 µg . 7.5% SDS PAGE. SHMT2 Polyclonal Antibody (Product # PA5-32228) diluted at 1:10,000.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: SHMT2 Polyclonal Antibody detects SHMT2 protein by Western blot analysis. A. 30 µg A431 whole cell lysate/extract. B. 30 µg HepG2 whole cell lysate/extract.7.5 % SDS-PAGE. SHMT2 Polyclonal Antibody (Product # PA5-32228) dilution: 1:5,000.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Knockdown of Serine hydroxymethyltransferase, mitochondrial was achieved by transfecting Hep G2 with Serine hydroxymethyltransferase, mitochondrial specific siRNAs (Silencer® select Product # s12825, s12823). Western blot analysis (Fig. a) was performed using Whole cell extracts from the Serine hydroxymethyltransferase, mitochondrial knockdown cells (lane 3), non-targeting scrambled siRNA transfected cells (lane 2) and untransfected cells (lane 1). The blot was probed with SHMT2 Polyclonal Antibody (Product # PA5-32228, 1:5000 dilution) and Goat anti-Rabbit IgG (H+L) Superclonal™ Recombinant Secondary Antibody, HRP (Product # A27036, 1:10,000 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 Serine hydroxymethyltransferase, mitochondrial.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Western blot was performed using Anti-SHMT2 Polyclonal Antibody (Product # PA5-32228) and a 55 kDa band corresponding to Serine hydroxymethyltransferase, mitochondrial was observed across the cell lines and tissues tested. Whole cell extracts (30 µg lysate) of A549 (Lane 1), HeLa (Lane 2), LNCaP (Lane 3), Hep G2 (Lane 4), MCF7 (Lane 5), MDA-MB-231 (Lane 6), U-87 MG (Lane 7), Mouse Skeletal Muscle (Lane 8), Rat Skeletal Muscle (Lane 9), Mouse Liver (Lane 10), Rat Liver (Lane 11) were electrophoresed using NuPAGE™ 4-12% Bis-Tris Protein Gel (Product # NP0321BOX). Resolved proteins were then transferred onto a Nitrocellulose membrane (Product # IB23001) by iBlot® 2 Dry Blotting System (Product # IB21001). The blot was probed with the primary antibody (1:5000 dilution) and detected by chemiluminescence with Goat anti-Rabbit IgG (H+L) Superclonal™ Recombinant Secondary Antibody, HRP (Product # A27036, 1:10,000 dilution) using the iBright FL 1000 (Product # A32752). Chemiluminescent detection was performed using Novex® ECL Chemiluminescent Substrate Reagent Kit (Product # WP20005).

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: SHMT2 Polyclonal Antibody detects SHMT2 protein at Mitochondria by immunofluorescent analysis. Sample: HeLa cells were fixed in ice-cold MeOH for 1 sec. Green: SHMT2 protein stained by SHMT2 Polyclonal Antibody (Product # PA5-32228) diluted at 1:500. Blue: Hoechst 33342 staining.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: SHMT2 Polyclonal Antibody detects SHMT2 protein at Mitochondria by immunofluorescent analysis. Sample: HeLa cells were fixed in ice-cold MeOH for 1 sec. Green: SHMT2 protein stained by SHMT2 Polyclonal Antibody (Product # PA5-32228) diluted at 1:500. Blue: Hoechst 33342 staining.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Immunofluorescence analysis of Serine hydroxymethyltransferase, mitochondrial 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 2% BSA for 45 minutes at room temperature. The cells were labeled with SHMT2 Polyclonal Antibody (Product # PA5-32228) 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™ Recombinant Secondary Antibody, Alexa Fluor® 488 conjugate (Product # A27034), (1:2000 dilution), 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, 1:300). 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: Immunohistochemical analysis of paraffin-embedded HeLa xenograft, using SHMT2 (Product # PA5-32228) 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: Figure 5--figure supplement 2. SHMT1 and SHMT2 single and double knockouts. ( a ) Western blots (top) and growth rates (bottom) for fourteen single cell clones derived from simultaneous CRISPR transfection for SHMT1 and SHMT2 knockout. All clones were expanded in the presence of hypoxanthine and thymidine (HT). ( b ) Western blots showing SHMT1 and SHMT2 single and double knockouts. ( c ) Formate synthesis from mitochondria isolated from SHMT1 and SHMT2 knockout cell lines. ( d ) Replicate serine dependence data from single knockouts. ( e ) Growth of SHMT1/SHMT2 double KO cells (two independent clones) with different 1C-related supplements. ( f ) 1C-rescued double knockouts do not show serine dependence, even when treated with RC inhibitor. n = 1 for panel a; n = 3 for panels c-f . DOI: http://dx.doi.org/

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Figure 7 Cellular redistribution of SHMT2 and PKM2 following IL-6 stimulation. Immunofluorescence analysis of SHMT2 ( A , B ) and PKM2 ( C , D ) proteins in LNCaP and DU145 cells after 4 h of IL-6 stimulation, with or without a Jak2 inhibitor (AZD1480). ( A , C ) LNCaP cell line. ( B , D ) DU145 cell line. The images were captured with a confocal fluorescence microscope (ZEISS LSM510META) using 60x oil immersion objective (scale bar 10 mum).

PA5-32228

Antibody data