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PDF1 | Plant defensin 1.1

AS16 3973 | Clonality: Polyclonal | Host: Rabbit | Reactivity: Solanum lycopersicum

PDF1 | Plant defensin 1.1 in the group Antibodies Plant/Algal  / Environmental Stress / Salt stress at Agrisera AB (Antibodies for research) (AS16 3973)
PDF1 | Plant defensin 1.1



DATA SHEET IN PDF

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Product Information

Immunogen

KLH-conjugated synthetic peptide derived from Arabidopsis thaliana PDF1.1 UniProt: P30224-1, TAIR: At1g75830

The peptide sequence, it is perfectly conserved in following Arabidopsis thaliana isoforms: PDF1.2c, PDF1.2b, PDF1.2A, PDF1.3. PDF2 isoform sequence did not come in the blast.

Host Rabbit
Clonality Polyclonal
Purity Affinity purified serum in PBS pH 7.4
Format Lyophilized
Quantity 50 µg
Reconstitution For reconstitution add 50 µl, of sterile water.
Storage

Store lyophilized/reconstituted at -20°C; once reconstituted make aliquots to avoid repeated freeze-thaw cycles. Please, remember to spin tubes briefly prior to opening them to avoid any losses that might occur from lyophilized material adhering to the cap or sides of the tubes.

Tested applications Western blot (WB)
Recommended dilution

1 : 1000 (WB)

Expected | apparent MW 8.7 kDa

Reactivity

Confirmed reactivity Arabidopsis thaliana, Solanim lycopersicum
Predicted reactivity Arabidopsis thaliana, Brassica rapa, Camelina sativa, Eutrema salsugineum, Capsella rubella, Sorghum sp.
Species of your interest not listed? Contact us
Not reactive in Nicotiana benthamiana, Solanum tuberosum

Application examples

Application examples Western blot using anti-PDF1 antibodies
Samples: 

Marker used is Prestained Protein SHARPMASS™ VI Protein MW marker (5-245 kDa) from Euroclone company.

sample 1:, 40 µg of Arabidopsis thaliana total protein from wildtype leaves 0 hour post infection with Botrytis cinerea (negative control);

sample 2: 40 µg of Arabidopsis thaliana total protein from mutant leaves (with induced expression of PDF1 mRNA) 0 hour post infection with Botrytis cinerea (positive control);

sample 3: 40 µg of Arabidopsis thaliana total protein from wild type leaves 24 hours post infection with Botrytis cinerea (positive control, PDF1 is expected to be induced by the pathogen used in the experiment);

sample 4: 40 µg of Arabidopsis thaliana total protein from mutant leaves (with induced expression of PDF1 mRNA) leaves 24 hours post infection with Botrytis cinerea (positive control);

samples "-", 10 µg of Arabidopsis thaliana total protein wild type seedlings grown in dark condition (Negative control)



Up to 40 µg/well of total protein extracted freshly from  6-week-old leaves of Arabidopsis thaliana with extraction buffer (125 mm Tris, pH 6.8, 4% [w/v] SDS, 20% [v/v] glycerol, 0.02% [w/v] bromophenol blue, 10% [v/v] β-mercaptoethanol) and denatured 95°C for 7 minutes were separated on a 4–20% Mini-PROTEAN® TGX™ Precast Protein Gels (biorad) SDS-PAGE  and blotted 1h to PVDF (pore size of   0.2 µm), using Trans-Blot Turbo Transfer System (Bio-Rad). Blot was blocked with 5% milk for 1h/RT with agitation. Blot was incubated in the primary antibody at a dilution of 1: 1 000 for overnight (~16hours) with agitation in PBS-T with agitation at +4 °C. The antibody solution was decanted and the blot was rinsed briefly twice, then washed once for 15 min and 3 times for 5 min in PBS-T at RT with agitation. Blot was incubated in Agrisera matching secondary antibody (anti-rabbit IgG horse radish peroxidase conjugated) diluted to 1:10 000 for 2h/RT with agitation. The blot was washed as above and developed for 3 min with Agrisera ECL SuperBright with ChemiDoc Imaging Systems (Bio-Rad). Exposure time was 10 seconds. 
Left membrane: Western blot detection. Right membrane: staining.

Courtesy Dr. Ricardo Lorrani, University of Rome Sapienza, Italy

Additional information

Related products

Background

Background PDF1 (Plant defensin 1)  provides broad-spectrum resistance to pathogens and possesses antifungal activity. Alternative names:  Anther-specific protein S18 homolog,Cysteine-rich antifungal protein 1, AFP1,Low-molecular-weight cysteine-rich protein 67, Protein LCR67.

Product citations

Selected references Nikoloudakis et al. (2020). Structural Diversity and Highly Specific Host-Pathogen Transcriptional Regulation of Defensin Genes Is Revealed in Tomato. Int J Mol Sci. 2020 Dec 9;21(24):9380. doi: 10.3390/ijms21249380. PMID: 33317090; PMCID: PMC7764197.
immunogen:

KLH-conjugated synthetic peptide derived from Arabidopsis thaliana PDF1.1 UniProt: P30224-1, TAIR: At1g75830

The peptide sequence, it is perfectly conserved in following Arabidopsis thaliana isoforms: PDF1.2c, PDF1.2b, PDF1.2A, PDF1.3. PDF2 isoform sequence did not come in the blast.

Reconstitution: For reconstitution add 50 µl, of sterile water.
Host: Rabbit
Clonality: Polyclonal
Purity: Affinity purified serum in PBS pH 7.4
Format: Lyophilized
Quantity: 50 µg
storage:

Store lyophilized/reconstituted at -20°C; once reconstituted make aliquots to avoid repeated freeze-thaw cycles. Please, remember to spin tubes briefly prior to opening them to avoid any losses that might occur from lyophilized material adhering to the cap or sides of the tubes.

tested applications: Western blot (WB)
recommended dilution:

1 : 1000 (WB)

calculated | apparent molecular mass [kDa]: 8.7 kDa
Confirmed reactivity: Arabidopsis thaliana, Solanim lycopersicum
predicted reactivity: Arabidopsis thaliana, Brassica rapa, Camelina sativa, Eutrema salsugineum, Capsella rubella, Sorghum sp.
Species of your interest not listed? Contact us
not reactive in: Nicotiana benthamiana, Solanum tuberosum
Picture (footer): Western blot using anti-PDF1 antibodies
Samples: 

Marker used is Prestained Protein SHARPMASS™ VI Protein MW marker (5-245 kDa) from Euroclone company.

sample 1:, 40 µg of Arabidopsis thaliana total protein from wildtype leaves 0 hour post infection with Botrytis cinerea (negative control);

sample 2: 40 µg of Arabidopsis thaliana total protein from mutant leaves (with induced expression of PDF1 mRNA) 0 hour post infection with Botrytis cinerea (positive control);

sample 3: 40 µg of Arabidopsis thaliana total protein from wild type leaves 24 hours post infection with Botrytis cinerea (positive control, PDF1 is expected to be induced by the pathogen used in the experiment);

sample 4: 40 µg of Arabidopsis thaliana total protein from mutant leaves (with induced expression of PDF1 mRNA) leaves 24 hours post infection with Botrytis cinerea (positive control);

samples "-", 10 µg of Arabidopsis thaliana total protein wild type seedlings grown in dark condition (Negative control)



Up to 40 µg/well of total protein extracted freshly from  6-week-old leaves of Arabidopsis thaliana with extraction buffer (125 mm Tris, pH 6.8, 4% [w/v] SDS, 20% [v/v] glycerol, 0.02% [w/v] bromophenol blue, 10% [v/v] β-mercaptoethanol) and denatured 95°C for 7 minutes were separated on a 4–20% Mini-PROTEAN® TGX™ Precast Protein Gels (biorad) SDS-PAGE  and blotted 1h to PVDF (pore size of   0.2 µm), using Trans-Blot Turbo Transfer System (Bio-Rad). Blot was blocked with 5% milk for 1h/RT with agitation. Blot was incubated in the primary antibody at a dilution of 1: 1 000 for overnight (~16hours) with agitation in PBS-T with agitation at +4 °C. The antibody solution was decanted and the blot was rinsed briefly twice, then washed once for 15 min and 3 times for 5 min in PBS-T at RT with agitation. Blot was incubated in Agrisera matching secondary antibody (anti-rabbit IgG horse radish peroxidase conjugated) diluted to 1:10 000 for 2h/RT with agitation. The blot was washed as above and developed for 3 min with Agrisera ECL SuperBright with ChemiDoc Imaging Systems (Bio-Rad). Exposure time was 10 seconds. 
Left membrane: Western blot detection. Right membrane: staining.

Courtesy Dr. Ricardo Lorrani, University of Rome Sapienza, Italy
background: PDF1 (Plant defensin 1)  provides broad-spectrum resistance to pathogens and possesses antifungal activity. Alternative names:  Anther-specific protein S18 homolog,Cysteine-rich antifungal protein 1, AFP1,Low-molecular-weight cysteine-rich protein 67, Protein LCR67.
All references: Nikoloudakis et al. (2020). Structural Diversity and Highly Specific Host-Pathogen Transcriptional Regulation of Defensin Genes Is Revealed in Tomato. Int J Mol Sci. 2020 Dec 9;21(24):9380. doi: 10.3390/ijms21249380. PMID: 33317090; PMCID: PMC7764197.

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