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rat anti-exp1  (ProteoGenix)

 
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    ProteoGenix rat anti-exp1
    Characterization of pPbU6-hdhfr/yfcu vector systems and the PbCas9 mother line. ( A ) Schematic of pPbU6-hdhfr/yfcu-Cas9 with the HRs of the repair template delivered on the same plasmid (one-plasmid approach) or on a PCR product (PCR-template approach) to generate the rap2/3 –3×HA parasite line. The effect on transfection efficiency was compared by both ( i ) substituting the PyU6 for the PbU6 promoter (not significant) and ( ii ) different homology arm length ( p = 0.0068). ( iii ) The relationship between homology arm length and transfection efficiency was examined by plotting homology arm length against the day parasitemia reached >0.5%. One or two-way ANOVAs were performed to compare transfection efficiencies from two biological replicates per condition. The expression of RAP2/3–3×HA was confirmed by western blot following transfection with the pPbU6-hdhfr/yfcu-Cas9 vector using ∼500-bp HR. Wild-type parasites were used as a negative control for the HA tag and the <t>anti-EXP1</t> was used as a loading control. ( C ) Growth assay of the PbCas9 line that constitutively expresses FLAG-Cas9 under the hsp70 promoter with the PbGIMO mother line used as a control. Error bars = SD for three biological replicates. ( D ) FLAG-Cas9 expression was confirmed using western blot, where FLAG detects Cas9 and anti-EXP1 was used as loading control. PbGIMO was used as a negative control for the FLAG tag. ( E ) IFA demonstrates that FLAG-Cas9 is expressed and localized to the nucleus. FLAG detects Cas9 and DAPI stains the nucleus. BF = Bright-field. The PbGIMO mother line was used as a negative control. Scale bar = 5 μm. ( F ) Transfection efficiency of the pPbU6-hdhfr/yfcu rap2/3 –3×HA vector into the PbCas9 line was assessed using the one-plasmid versus the PCR-template approach. Error bars = SD for two biological replicates. Full-length western blots are shown in and and parasitemia counts can be found in .
    Rat Anti Exp1, supplied by ProteoGenix, used in various techniques. Bioz Stars score: 90/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/product/rat+anti-exp1/rat+anti+exp1/pmc11754126-180-23-28
    Average 90 stars, based on 1 article reviews
    rat anti-exp1 - by Bioz Stars, 2026-10
    90/100 stars

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    1) Product Images from "A scalable CRISPR-Cas9 gene editing system facilitates CRISPR screens in the malaria parasite Plasmodium berghei"

    Article Title: A scalable CRISPR-Cas9 gene editing system facilitates CRISPR screens in the malaria parasite Plasmodium berghei

    Journal: Nucleic Acids Research

    doi: 10.1093/nar/gkaf005

    Characterization of pPbU6-hdhfr/yfcu vector systems and the PbCas9 mother line. ( A ) Schematic of pPbU6-hdhfr/yfcu-Cas9 with the HRs of the repair template delivered on the same plasmid (one-plasmid approach) or on a PCR product (PCR-template approach) to generate the rap2/3 –3×HA parasite line. The effect on transfection efficiency was compared by both ( i ) substituting the PyU6 for the PbU6 promoter (not significant) and ( ii ) different homology arm length ( p = 0.0068). ( iii ) The relationship between homology arm length and transfection efficiency was examined by plotting homology arm length against the day parasitemia reached >0.5%. One or two-way ANOVAs were performed to compare transfection efficiencies from two biological replicates per condition. The expression of RAP2/3–3×HA was confirmed by western blot following transfection with the pPbU6-hdhfr/yfcu-Cas9 vector using ∼500-bp HR. Wild-type parasites were used as a negative control for the HA tag and the anti-EXP1 was used as a loading control. ( C ) Growth assay of the PbCas9 line that constitutively expresses FLAG-Cas9 under the hsp70 promoter with the PbGIMO mother line used as a control. Error bars = SD for three biological replicates. ( D ) FLAG-Cas9 expression was confirmed using western blot, where FLAG detects Cas9 and anti-EXP1 was used as loading control. PbGIMO was used as a negative control for the FLAG tag. ( E ) IFA demonstrates that FLAG-Cas9 is expressed and localized to the nucleus. FLAG detects Cas9 and DAPI stains the nucleus. BF = Bright-field. The PbGIMO mother line was used as a negative control. Scale bar = 5 μm. ( F ) Transfection efficiency of the pPbU6-hdhfr/yfcu rap2/3 –3×HA vector into the PbCas9 line was assessed using the one-plasmid versus the PCR-template approach. Error bars = SD for two biological replicates. Full-length western blots are shown in and and parasitemia counts can be found in .
    Figure Legend Snippet: Characterization of pPbU6-hdhfr/yfcu vector systems and the PbCas9 mother line. ( A ) Schematic of pPbU6-hdhfr/yfcu-Cas9 with the HRs of the repair template delivered on the same plasmid (one-plasmid approach) or on a PCR product (PCR-template approach) to generate the rap2/3 –3×HA parasite line. The effect on transfection efficiency was compared by both ( i ) substituting the PyU6 for the PbU6 promoter (not significant) and ( ii ) different homology arm length ( p = 0.0068). ( iii ) The relationship between homology arm length and transfection efficiency was examined by plotting homology arm length against the day parasitemia reached >0.5%. One or two-way ANOVAs were performed to compare transfection efficiencies from two biological replicates per condition. The expression of RAP2/3–3×HA was confirmed by western blot following transfection with the pPbU6-hdhfr/yfcu-Cas9 vector using ∼500-bp HR. Wild-type parasites were used as a negative control for the HA tag and the anti-EXP1 was used as a loading control. ( C ) Growth assay of the PbCas9 line that constitutively expresses FLAG-Cas9 under the hsp70 promoter with the PbGIMO mother line used as a control. Error bars = SD for three biological replicates. ( D ) FLAG-Cas9 expression was confirmed using western blot, where FLAG detects Cas9 and anti-EXP1 was used as loading control. PbGIMO was used as a negative control for the FLAG tag. ( E ) IFA demonstrates that FLAG-Cas9 is expressed and localized to the nucleus. FLAG detects Cas9 and DAPI stains the nucleus. BF = Bright-field. The PbGIMO mother line was used as a negative control. Scale bar = 5 μm. ( F ) Transfection efficiency of the pPbU6-hdhfr/yfcu rap2/3 –3×HA vector into the PbCas9 line was assessed using the one-plasmid versus the PCR-template approach. Error bars = SD for two biological replicates. Full-length western blots are shown in and and parasitemia counts can be found in .

    Techniques Used: Plasmid Preparation, Transfection, Expressing, Western Blot, Negative Control, Control, Growth Assay, FLAG-tag

    Efficient epitope tagging using the PbHiT system. ( A ) Schematic overview of pPbHiT plasmid used for cMyc epitope tagging of the gene of interest. ( i ) Daily parasitemia counts post-transfection comparing 50- and 100-bp homology arms for rap2/3 and PBANKA_1224200 tagging vectors using a single gRNA. For both genes, the 100-bp HR results in faster recovery of transgenic parasites. Error bars = SD for two biological replicates. ( ii ) Daily parasitemia counts post-transfection comparing transfection of six individual genes targeted (PBANKA_1224200, PBANKA_1225600, PBANKA_0914500, PBANKA_0622900, PBANKA_1451000 and PBANKA_0829400) using 100-bp homology arms and two gRNAs (G1 and G2) per target except for PBANKA_0829400. Both the gene targeted and the gRNA used affect transfection efficiency. All parasitemia counts can be found in . ( B ) The expression of tagged proteins was confirmed using western blot using anti-HA antibody. The PbCas9 background line was used as a negative control and anti-EXP1 or PfHSP70 were used as loading controls. Full-length western blots are shown in . ( C ) IFAs of both RAP2/3–3×cMyc and PBANKA_0622900–3×cMyc showed that both proteins are expressed during the schizont stage as expected. Anti-EXP1 was used as a marker for the parasitophorous vacuole membrane/dense granules and DAPI to stain the nucleus. Scale bars = 5 μm.
    Figure Legend Snippet: Efficient epitope tagging using the PbHiT system. ( A ) Schematic overview of pPbHiT plasmid used for cMyc epitope tagging of the gene of interest. ( i ) Daily parasitemia counts post-transfection comparing 50- and 100-bp homology arms for rap2/3 and PBANKA_1224200 tagging vectors using a single gRNA. For both genes, the 100-bp HR results in faster recovery of transgenic parasites. Error bars = SD for two biological replicates. ( ii ) Daily parasitemia counts post-transfection comparing transfection of six individual genes targeted (PBANKA_1224200, PBANKA_1225600, PBANKA_0914500, PBANKA_0622900, PBANKA_1451000 and PBANKA_0829400) using 100-bp homology arms and two gRNAs (G1 and G2) per target except for PBANKA_0829400. Both the gene targeted and the gRNA used affect transfection efficiency. All parasitemia counts can be found in . ( B ) The expression of tagged proteins was confirmed using western blot using anti-HA antibody. The PbCas9 background line was used as a negative control and anti-EXP1 or PfHSP70 were used as loading controls. Full-length western blots are shown in . ( C ) IFAs of both RAP2/3–3×cMyc and PBANKA_0622900–3×cMyc showed that both proteins are expressed during the schizont stage as expected. Anti-EXP1 was used as a marker for the parasitophorous vacuole membrane/dense granules and DAPI to stain the nucleus. Scale bars = 5 μm.

    Techniques Used: Plasmid Preparation, Transfection, Transgenic Assay, Expressing, Western Blot, Negative Control, Marker, Membrane, Staining

    Related Articles

    Plasmid Preparation:

    Article Title: A scalable CRISPR-Cas9 gene editing system facilitates CRISPR screens in the malaria parasite Plasmodium berghei
    Article Snippet: Primary antibodies used were as follows: rabbit anti-FLAG (Sigma–Aldrich #SAB4301135, 1:1000), rabbit anti-cMyc (Cell Signal #2278, 1:1000), rabbit anti-HA (Cell Signal #C29F4, 1:1000), rat anti-EXP1 (custom made from Proteogenix, 1:1000) and rabbit anti-PfHSP70 (LS Bio #LS-C109068-25, 1:10 000).

    Article Title: A scalable CRISPR-Cas9 gene editing system facilitates CRISPR screens in the malaria parasite Plasmodium berghei.
    Article Snippet: Primary antibodies used were as follows: rabbit anti-FLAG (Sigma– Aldrich #SAB4301135, 1:1000), rabbit anti-cMyc (Cell Signal #2278, 1:1000), rabbit anti-HA (Cell Signal #C29F4, 1:1000), rat anti-EXP1 (custom made from Proteogenix, 1:1000) and rabbit anti-PfHSP70 (LS Bio #LS-C109068-25, 1:10 000).

    Transfection:

    Article Title: A scalable CRISPR-Cas9 gene editing system facilitates CRISPR screens in the malaria parasite Plasmodium berghei
    Article Snippet: Primary antibodies used were as follows: rabbit anti-FLAG (Sigma–Aldrich #SAB4301135, 1:1000), rabbit anti-cMyc (Cell Signal #2278, 1:1000), rabbit anti-HA (Cell Signal #C29F4, 1:1000), rat anti-EXP1 (custom made from Proteogenix, 1:1000) and rabbit anti-PfHSP70 (LS Bio #LS-C109068-25, 1:10 000).

    Article Title: A scalable CRISPR-Cas9 gene editing system facilitates CRISPR screens in the malaria parasite Plasmodium berghei.
    Article Snippet: Primary antibodies used were as follows: rabbit anti-FLAG (Sigma– Aldrich #SAB4301135, 1:1000), rabbit anti-cMyc (Cell Signal #2278, 1:1000), rabbit anti-HA (Cell Signal #C29F4, 1:1000), rat anti-EXP1 (custom made from Proteogenix, 1:1000) and rabbit anti-PfHSP70 (LS Bio #LS-C109068-25, 1:10 000).

    Expressing:

    Article Title: A scalable CRISPR-Cas9 gene editing system facilitates CRISPR screens in the malaria parasite Plasmodium berghei
    Article Snippet: Primary antibodies used were as follows: rabbit anti-FLAG (Sigma–Aldrich #SAB4301135, 1:1000), rabbit anti-cMyc (Cell Signal #2278, 1:1000), rabbit anti-HA (Cell Signal #C29F4, 1:1000), rat anti-EXP1 (custom made from Proteogenix, 1:1000) and rabbit anti-PfHSP70 (LS Bio #LS-C109068-25, 1:10 000).

    Article Title: A scalable CRISPR-Cas9 gene editing system facilitates CRISPR screens in the malaria parasite Plasmodium berghei.
    Article Snippet: Primary antibodies used were as follows: rabbit anti-FLAG (Sigma– Aldrich #SAB4301135, 1:1000), rabbit anti-cMyc (Cell Signal #2278, 1:1000), rabbit anti-HA (Cell Signal #C29F4, 1:1000), rat anti-EXP1 (custom made from Proteogenix, 1:1000) and rabbit anti-PfHSP70 (LS Bio #LS-C109068-25, 1:10 000).

    Western Blot:

    Article Title: A scalable CRISPR-Cas9 gene editing system facilitates CRISPR screens in the malaria parasite Plasmodium berghei
    Article Snippet: Primary antibodies used were as follows: rabbit anti-FLAG (Sigma–Aldrich #SAB4301135, 1:1000), rabbit anti-cMyc (Cell Signal #2278, 1:1000), rabbit anti-HA (Cell Signal #C29F4, 1:1000), rat anti-EXP1 (custom made from Proteogenix, 1:1000) and rabbit anti-PfHSP70 (LS Bio #LS-C109068-25, 1:10 000).

    Article Title: A scalable CRISPR-Cas9 gene editing system facilitates CRISPR screens in the malaria parasite Plasmodium berghei.
    Article Snippet: Primary antibodies used were as follows: rabbit anti-FLAG (Sigma– Aldrich #SAB4301135, 1:1000), rabbit anti-cMyc (Cell Signal #2278, 1:1000), rabbit anti-HA (Cell Signal #C29F4, 1:1000), rat anti-EXP1 (custom made from Proteogenix, 1:1000) and rabbit anti-PfHSP70 (LS Bio #LS-C109068-25, 1:10 000).

    Negative Control:

    Article Title: A scalable CRISPR-Cas9 gene editing system facilitates CRISPR screens in the malaria parasite Plasmodium berghei
    Article Snippet: Primary antibodies used were as follows: rabbit anti-FLAG (Sigma–Aldrich #SAB4301135, 1:1000), rabbit anti-cMyc (Cell Signal #2278, 1:1000), rabbit anti-HA (Cell Signal #C29F4, 1:1000), rat anti-EXP1 (custom made from Proteogenix, 1:1000) and rabbit anti-PfHSP70 (LS Bio #LS-C109068-25, 1:10 000).

    Article Title: A scalable CRISPR-Cas9 gene editing system facilitates CRISPR screens in the malaria parasite Plasmodium berghei.
    Article Snippet: Primary antibodies used were as follows: rabbit anti-FLAG (Sigma– Aldrich #SAB4301135, 1:1000), rabbit anti-cMyc (Cell Signal #2278, 1:1000), rabbit anti-HA (Cell Signal #C29F4, 1:1000), rat anti-EXP1 (custom made from Proteogenix, 1:1000) and rabbit anti-PfHSP70 (LS Bio #LS-C109068-25, 1:10 000).

    Control:

    Article Title: A scalable CRISPR-Cas9 gene editing system facilitates CRISPR screens in the malaria parasite Plasmodium berghei
    Article Snippet: Primary antibodies used were as follows: rabbit anti-FLAG (Sigma–Aldrich #SAB4301135, 1:1000), rabbit anti-cMyc (Cell Signal #2278, 1:1000), rabbit anti-HA (Cell Signal #C29F4, 1:1000), rat anti-EXP1 (custom made from Proteogenix, 1:1000) and rabbit anti-PfHSP70 (LS Bio #LS-C109068-25, 1:10 000).

    Article Title: A scalable CRISPR-Cas9 gene editing system facilitates CRISPR screens in the malaria parasite Plasmodium berghei.
    Article Snippet: Primary antibodies used were as follows: rabbit anti-FLAG (Sigma– Aldrich #SAB4301135, 1:1000), rabbit anti-cMyc (Cell Signal #2278, 1:1000), rabbit anti-HA (Cell Signal #C29F4, 1:1000), rat anti-EXP1 (custom made from Proteogenix, 1:1000) and rabbit anti-PfHSP70 (LS Bio #LS-C109068-25, 1:10 000).

    Growth Assay:

    Article Title: A scalable CRISPR-Cas9 gene editing system facilitates CRISPR screens in the malaria parasite Plasmodium berghei
    Article Snippet: Primary antibodies used were as follows: rabbit anti-FLAG (Sigma–Aldrich #SAB4301135, 1:1000), rabbit anti-cMyc (Cell Signal #2278, 1:1000), rabbit anti-HA (Cell Signal #C29F4, 1:1000), rat anti-EXP1 (custom made from Proteogenix, 1:1000) and rabbit anti-PfHSP70 (LS Bio #LS-C109068-25, 1:10 000).

    Article Title: A scalable CRISPR-Cas9 gene editing system facilitates CRISPR screens in the malaria parasite Plasmodium berghei.
    Article Snippet: Primary antibodies used were as follows: rabbit anti-FLAG (Sigma– Aldrich #SAB4301135, 1:1000), rabbit anti-cMyc (Cell Signal #2278, 1:1000), rabbit anti-HA (Cell Signal #C29F4, 1:1000), rat anti-EXP1 (custom made from Proteogenix, 1:1000) and rabbit anti-PfHSP70 (LS Bio #LS-C109068-25, 1:10 000).

    FLAG-tag:

    Article Title: A scalable CRISPR-Cas9 gene editing system facilitates CRISPR screens in the malaria parasite Plasmodium berghei
    Article Snippet: Primary antibodies used were as follows: rabbit anti-FLAG (Sigma–Aldrich #SAB4301135, 1:1000), rabbit anti-cMyc (Cell Signal #2278, 1:1000), rabbit anti-HA (Cell Signal #C29F4, 1:1000), rat anti-EXP1 (custom made from Proteogenix, 1:1000) and rabbit anti-PfHSP70 (LS Bio #LS-C109068-25, 1:10 000).

    Article Title: A scalable CRISPR-Cas9 gene editing system facilitates CRISPR screens in the malaria parasite Plasmodium berghei.
    Article Snippet: Primary antibodies used were as follows: rabbit anti-FLAG (Sigma– Aldrich #SAB4301135, 1:1000), rabbit anti-cMyc (Cell Signal #2278, 1:1000), rabbit anti-HA (Cell Signal #C29F4, 1:1000), rat anti-EXP1 (custom made from Proteogenix, 1:1000) and rabbit anti-PfHSP70 (LS Bio #LS-C109068-25, 1:10 000).

    Transgenic Assay:

    Article Title: A scalable CRISPR-Cas9 gene editing system facilitates CRISPR screens in the malaria parasite Plasmodium berghei
    Article Snippet: Primary antibodies used were as follows: rabbit anti-FLAG (Sigma–Aldrich #SAB4301135, 1:1000), rabbit anti-cMyc (Cell Signal #2278, 1:1000), rabbit anti-HA (Cell Signal #C29F4, 1:1000), rat anti-EXP1 (custom made from Proteogenix, 1:1000) and rabbit anti-PfHSP70 (LS Bio #LS-C109068-25, 1:10 000).

    Article Title: A scalable CRISPR-Cas9 gene editing system facilitates CRISPR screens in the malaria parasite Plasmodium berghei.
    Article Snippet: Primary antibodies used were as follows: rabbit anti-FLAG (Sigma– Aldrich #SAB4301135, 1:1000), rabbit anti-cMyc (Cell Signal #2278, 1:1000), rabbit anti-HA (Cell Signal #C29F4, 1:1000), rat anti-EXP1 (custom made from Proteogenix, 1:1000) and rabbit anti-PfHSP70 (LS Bio #LS-C109068-25, 1:10 000).

    Marker:

    Article Title: A scalable CRISPR-Cas9 gene editing system facilitates CRISPR screens in the malaria parasite Plasmodium berghei
    Article Snippet: Primary antibodies used were as follows: rabbit anti-FLAG (Sigma–Aldrich #SAB4301135, 1:1000), rabbit anti-cMyc (Cell Signal #2278, 1:1000), rabbit anti-HA (Cell Signal #C29F4, 1:1000), rat anti-EXP1 (custom made from Proteogenix, 1:1000) and rabbit anti-PfHSP70 (LS Bio #LS-C109068-25, 1:10 000).

    Article Title: A scalable CRISPR-Cas9 gene editing system facilitates CRISPR screens in the malaria parasite Plasmodium berghei.
    Article Snippet: Primary antibodies used were as follows: rabbit anti-FLAG (Sigma– Aldrich #SAB4301135, 1:1000), rabbit anti-cMyc (Cell Signal #2278, 1:1000), rabbit anti-HA (Cell Signal #C29F4, 1:1000), rat anti-EXP1 (custom made from Proteogenix, 1:1000) and rabbit anti-PfHSP70 (LS Bio #LS-C109068-25, 1:10 000).

    Membrane:

    Article Title: A scalable CRISPR-Cas9 gene editing system facilitates CRISPR screens in the malaria parasite Plasmodium berghei
    Article Snippet: Primary antibodies used were as follows: rabbit anti-FLAG (Sigma–Aldrich #SAB4301135, 1:1000), rabbit anti-cMyc (Cell Signal #2278, 1:1000), rabbit anti-HA (Cell Signal #C29F4, 1:1000), rat anti-EXP1 (custom made from Proteogenix, 1:1000) and rabbit anti-PfHSP70 (LS Bio #LS-C109068-25, 1:10 000).

    Article Title: A scalable CRISPR-Cas9 gene editing system facilitates CRISPR screens in the malaria parasite Plasmodium berghei.
    Article Snippet: Primary antibodies used were as follows: rabbit anti-FLAG (Sigma– Aldrich #SAB4301135, 1:1000), rabbit anti-cMyc (Cell Signal #2278, 1:1000), rabbit anti-HA (Cell Signal #C29F4, 1:1000), rat anti-EXP1 (custom made from Proteogenix, 1:1000) and rabbit anti-PfHSP70 (LS Bio #LS-C109068-25, 1:10 000).

    Staining:

    Article Title: A scalable CRISPR-Cas9 gene editing system facilitates CRISPR screens in the malaria parasite Plasmodium berghei
    Article Snippet: Primary antibodies used were as follows: rabbit anti-FLAG (Sigma–Aldrich #SAB4301135, 1:1000), rabbit anti-cMyc (Cell Signal #2278, 1:1000), rabbit anti-HA (Cell Signal #C29F4, 1:1000), rat anti-EXP1 (custom made from Proteogenix, 1:1000) and rabbit anti-PfHSP70 (LS Bio #LS-C109068-25, 1:10 000).

    Article Title: A scalable CRISPR-Cas9 gene editing system facilitates CRISPR screens in the malaria parasite Plasmodium berghei.
    Article Snippet: Primary antibodies used were as follows: rabbit anti-FLAG (Sigma– Aldrich #SAB4301135, 1:1000), rabbit anti-cMyc (Cell Signal #2278, 1:1000), rabbit anti-HA (Cell Signal #C29F4, 1:1000), rat anti-EXP1 (custom made from Proteogenix, 1:1000) and rabbit anti-PfHSP70 (LS Bio #LS-C109068-25, 1:10 000).



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    rat anti-exp1  (ProteoGenix)
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    ProteoGenix rat anti-exp1
    Characterization of pPbU6-hdhfr/yfcu vector systems and the PbCas9 mother line. ( A ) Schematic of pPbU6-hdhfr/yfcu-Cas9 with the HRs of the repair template delivered on the same plasmid (one-plasmid approach) or on a PCR product (PCR-template approach) to generate the rap2/3 –3×HA parasite line. The effect on transfection efficiency was compared by both ( i ) substituting the PyU6 for the PbU6 promoter (not significant) and ( ii ) different homology arm length ( p = 0.0068). ( iii ) The relationship between homology arm length and transfection efficiency was examined by plotting homology arm length against the day parasitemia reached >0.5%. One or two-way ANOVAs were performed to compare transfection efficiencies from two biological replicates per condition. The expression of RAP2/3–3×HA was confirmed by western blot following transfection with the pPbU6-hdhfr/yfcu-Cas9 vector using ∼500-bp HR. Wild-type parasites were used as a negative control for the HA tag and the <t>anti-EXP1</t> was used as a loading control. ( C ) Growth assay of the PbCas9 line that constitutively expresses FLAG-Cas9 under the hsp70 promoter with the PbGIMO mother line used as a control. Error bars = SD for three biological replicates. ( D ) FLAG-Cas9 expression was confirmed using western blot, where FLAG detects Cas9 and anti-EXP1 was used as loading control. PbGIMO was used as a negative control for the FLAG tag. ( E ) IFA demonstrates that FLAG-Cas9 is expressed and localized to the nucleus. FLAG detects Cas9 and DAPI stains the nucleus. BF = Bright-field. The PbGIMO mother line was used as a negative control. Scale bar = 5 μm. ( F ) Transfection efficiency of the pPbU6-hdhfr/yfcu rap2/3 –3×HA vector into the PbCas9 line was assessed using the one-plasmid versus the PCR-template approach. Error bars = SD for two biological replicates. Full-length western blots are shown in and and parasitemia counts can be found in .
    Rat Anti Exp1, supplied by ProteoGenix, used in various techniques. Bioz Stars score: 90/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/product/rat+anti-exp1/rat+anti+exp1/pmc11754126-180-23-28
    Average 90 stars, based on 1 article reviews
    rat anti-exp1 - by Bioz Stars, 2026-10
    90/100 stars
    		  Buy from Supplier

    Image Search Results


    Characterization of pPbU6-hdhfr/yfcu vector systems and the PbCas9 mother line. ( A ) Schematic of pPbU6-hdhfr/yfcu-Cas9 with the HRs of the repair template delivered on the same plasmid (one-plasmid approach) or on a PCR product (PCR-template approach) to generate the rap2/3 –3×HA parasite line. The effect on transfection efficiency was compared by both ( i ) substituting the PyU6 for the PbU6 promoter (not significant) and ( ii ) different homology arm length ( p = 0.0068). ( iii ) The relationship between homology arm length and transfection efficiency was examined by plotting homology arm length against the day parasitemia reached >0.5%. One or two-way ANOVAs were performed to compare transfection efficiencies from two biological replicates per condition. The expression of RAP2/3–3×HA was confirmed by western blot following transfection with the pPbU6-hdhfr/yfcu-Cas9 vector using ∼500-bp HR. Wild-type parasites were used as a negative control for the HA tag and the anti-EXP1 was used as a loading control. ( C ) Growth assay of the PbCas9 line that constitutively expresses FLAG-Cas9 under the hsp70 promoter with the PbGIMO mother line used as a control. Error bars = SD for three biological replicates. ( D ) FLAG-Cas9 expression was confirmed using western blot, where FLAG detects Cas9 and anti-EXP1 was used as loading control. PbGIMO was used as a negative control for the FLAG tag. ( E ) IFA demonstrates that FLAG-Cas9 is expressed and localized to the nucleus. FLAG detects Cas9 and DAPI stains the nucleus. BF = Bright-field. The PbGIMO mother line was used as a negative control. Scale bar = 5 μm. ( F ) Transfection efficiency of the pPbU6-hdhfr/yfcu rap2/3 –3×HA vector into the PbCas9 line was assessed using the one-plasmid versus the PCR-template approach. Error bars = SD for two biological replicates. Full-length western blots are shown in and and parasitemia counts can be found in .

    Journal: Nucleic Acids Research

    Article Title: A scalable CRISPR-Cas9 gene editing system facilitates CRISPR screens in the malaria parasite Plasmodium berghei

    doi: 10.1093/nar/gkaf005

    Figure Lengend Snippet: Characterization of pPbU6-hdhfr/yfcu vector systems and the PbCas9 mother line. ( A ) Schematic of pPbU6-hdhfr/yfcu-Cas9 with the HRs of the repair template delivered on the same plasmid (one-plasmid approach) or on a PCR product (PCR-template approach) to generate the rap2/3 –3×HA parasite line. The effect on transfection efficiency was compared by both ( i ) substituting the PyU6 for the PbU6 promoter (not significant) and ( ii ) different homology arm length ( p = 0.0068). ( iii ) The relationship between homology arm length and transfection efficiency was examined by plotting homology arm length against the day parasitemia reached >0.5%. One or two-way ANOVAs were performed to compare transfection efficiencies from two biological replicates per condition. The expression of RAP2/3–3×HA was confirmed by western blot following transfection with the pPbU6-hdhfr/yfcu-Cas9 vector using ∼500-bp HR. Wild-type parasites were used as a negative control for the HA tag and the anti-EXP1 was used as a loading control. ( C ) Growth assay of the PbCas9 line that constitutively expresses FLAG-Cas9 under the hsp70 promoter with the PbGIMO mother line used as a control. Error bars = SD for three biological replicates. ( D ) FLAG-Cas9 expression was confirmed using western blot, where FLAG detects Cas9 and anti-EXP1 was used as loading control. PbGIMO was used as a negative control for the FLAG tag. ( E ) IFA demonstrates that FLAG-Cas9 is expressed and localized to the nucleus. FLAG detects Cas9 and DAPI stains the nucleus. BF = Bright-field. The PbGIMO mother line was used as a negative control. Scale bar = 5 μm. ( F ) Transfection efficiency of the pPbU6-hdhfr/yfcu rap2/3 –3×HA vector into the PbCas9 line was assessed using the one-plasmid versus the PCR-template approach. Error bars = SD for two biological replicates. Full-length western blots are shown in and and parasitemia counts can be found in .

    Article Snippet: Primary antibodies used were as follows: rabbit anti-FLAG (Sigma–Aldrich #SAB4301135, 1:1000), rabbit anti-cMyc (Cell Signal #2278, 1:1000), rabbit anti-HA (Cell Signal #C29F4, 1:1000), rat anti-EXP1 (custom made from Proteogenix, 1:1000) and rabbit anti-PfHSP70 (LS Bio #LS-C109068-25, 1:10 000).

    Techniques: Plasmid Preparation, Transfection, Expressing, Western Blot, Negative Control, Control, Growth Assay, FLAG-tag

    Efficient epitope tagging using the PbHiT system. ( A ) Schematic overview of pPbHiT plasmid used for cMyc epitope tagging of the gene of interest. ( i ) Daily parasitemia counts post-transfection comparing 50- and 100-bp homology arms for rap2/3 and PBANKA_1224200 tagging vectors using a single gRNA. For both genes, the 100-bp HR results in faster recovery of transgenic parasites. Error bars = SD for two biological replicates. ( ii ) Daily parasitemia counts post-transfection comparing transfection of six individual genes targeted (PBANKA_1224200, PBANKA_1225600, PBANKA_0914500, PBANKA_0622900, PBANKA_1451000 and PBANKA_0829400) using 100-bp homology arms and two gRNAs (G1 and G2) per target except for PBANKA_0829400. Both the gene targeted and the gRNA used affect transfection efficiency. All parasitemia counts can be found in . ( B ) The expression of tagged proteins was confirmed using western blot using anti-HA antibody. The PbCas9 background line was used as a negative control and anti-EXP1 or PfHSP70 were used as loading controls. Full-length western blots are shown in . ( C ) IFAs of both RAP2/3–3×cMyc and PBANKA_0622900–3×cMyc showed that both proteins are expressed during the schizont stage as expected. Anti-EXP1 was used as a marker for the parasitophorous vacuole membrane/dense granules and DAPI to stain the nucleus. Scale bars = 5 μm.

    Journal: Nucleic Acids Research

    Article Title: A scalable CRISPR-Cas9 gene editing system facilitates CRISPR screens in the malaria parasite Plasmodium berghei

    doi: 10.1093/nar/gkaf005

    Figure Lengend Snippet: Efficient epitope tagging using the PbHiT system. ( A ) Schematic overview of pPbHiT plasmid used for cMyc epitope tagging of the gene of interest. ( i ) Daily parasitemia counts post-transfection comparing 50- and 100-bp homology arms for rap2/3 and PBANKA_1224200 tagging vectors using a single gRNA. For both genes, the 100-bp HR results in faster recovery of transgenic parasites. Error bars = SD for two biological replicates. ( ii ) Daily parasitemia counts post-transfection comparing transfection of six individual genes targeted (PBANKA_1224200, PBANKA_1225600, PBANKA_0914500, PBANKA_0622900, PBANKA_1451000 and PBANKA_0829400) using 100-bp homology arms and two gRNAs (G1 and G2) per target except for PBANKA_0829400. Both the gene targeted and the gRNA used affect transfection efficiency. All parasitemia counts can be found in . ( B ) The expression of tagged proteins was confirmed using western blot using anti-HA antibody. The PbCas9 background line was used as a negative control and anti-EXP1 or PfHSP70 were used as loading controls. Full-length western blots are shown in . ( C ) IFAs of both RAP2/3–3×cMyc and PBANKA_0622900–3×cMyc showed that both proteins are expressed during the schizont stage as expected. Anti-EXP1 was used as a marker for the parasitophorous vacuole membrane/dense granules and DAPI to stain the nucleus. Scale bars = 5 μm.

    Article Snippet: Primary antibodies used were as follows: rabbit anti-FLAG (Sigma–Aldrich #SAB4301135, 1:1000), rabbit anti-cMyc (Cell Signal #2278, 1:1000), rabbit anti-HA (Cell Signal #C29F4, 1:1000), rat anti-EXP1 (custom made from Proteogenix, 1:1000) and rabbit anti-PfHSP70 (LS Bio #LS-C109068-25, 1:10 000).

    Techniques: Plasmid Preparation, Transfection, Transgenic Assay, Expressing, Western Blot, Negative Control, Marker, Membrane, Staining