rabbit anti ha  (Cell Signaling Technology Inc)

 
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    Name:
    Anti rabbit IgG H L Biotinylated Antibody
    Description:
    Affinity purified goat anti rabbit IgG H L antibody is conjugated to biotin This product has been optimized for use as a secondary antibody in western blotting applications
    Catalog Number:
    14708
    Price:
    None
    Category:
    Secondary Antibodies
    Applications:
    Western Blot
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    Structured Review

    Cell Signaling Technology Inc rabbit anti ha
    Affinity purified goat anti rabbit IgG H L antibody is conjugated to biotin This product has been optimized for use as a secondary antibody in western blotting applications
    https://www.bioz.com/result/rabbit anti ha/product/Cell Signaling Technology Inc
    Average 86 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    rabbit anti ha - by Bioz Stars, 2021-07
    86/100 stars

    Images

    Related Articles

    Immunoprecipitation:

    Article Title: Novel Regulatory Roles of Wnt1 in Infection-Associated Colorectal Cancer
    Article Snippet: .. Anti-Wnt1 was not immunoprecipitated by anti-IgG ( A , right panel ). ..

    Article Title: Novel Regulatory Roles of Wnt1 in Infection-Associated Colorectal Cancer
    Article Snippet: .. To evaluate ubiquitination of Wnt1, we did immunoprecipitation using anti-ubiquitin, anti-Wnt1, or anti-IgG (negative control) antibodies, respectively. ..

    Article Title: MicroRNAs Overcome Cell Fate Barrier by Reducing EZH2-Controlled REST Stability during Neuronal Conversion of Human Adult Fibroblasts
    Article Snippet: Crude nuclei were isolated in SONIC buffer (50mM HEPES (pH7.6), 140mM NaCl, 1mM EDTA, 1% Triton X-100, 0.1% Na-deoxyxhilate and 0.1% SDS) and were sonicated with probe-based sonicator. .. The sheared chromatin samples were immunoprecipitated overnight at 4 °C under gentle rotation with anti-IgG (CST, #2729), or anti-REST (Bethyl, A300-539) antibody, and then add ChIP-grade protein G magnetic beads to each IP reaction and incubate for 2h at 4 °C with rotati on. ..

    other:

    Article Title: Neuregulin-1 attenuates experimental cerebral malaria (ECM) pathogenesis by regulating ErbB4/AKT/STAT3 signaling
    Article Snippet: Rabbit antibody against STAT3, phospho-STAT3, rabbit antibody against AKT, phospho-AKT, anti-NeuN, anti-Iba1, and anti-glial fibrillary acidic protein (GFAP) were purchased from Cell Signaling Technology Inc. (Danvers, MA, USA).

    Chromatin Immunoprecipitation:

    Article Title: Toxoplasma gondii excreted‐secreted antigens suppress Foxp3 promoter activity via a SP1‐dependent mechanism. Toxoplasma gondii excreted‐secreted antigens suppress Foxp3 promoter activity via a SP1‐dependent mechanism
    Article Snippet: The chromatin was harvested and fragmented using enzymatic digestion and sonication. .. Chromatin immunoprecipitation was performed with anti‐histone H3 (CST), anti‐SP1 (Santa Cruz Biotechnology) or anti‐IgG (CST). .. Anti‐histone H3 antibody and mouse IgG were utilized as positive and negative control, respectively.

    Article Title: Ferroptosis is governed by differential regulation of transcription in liver cancer
    Article Snippet: The antibodies used for co-IP were anti-HIC1 (Sigma, #SAB1412231, and #SAB2103167) and anti-HNF4A (R & D Systems, PP-H6939-00), and the conventional co-IP protocols are available elsewhere. .. The antibodies used for ChIP were anti-H3K9Ac (Abcam, #ab4441), anti-KAT2B (Abcam, #ab12188), anti-HNF4A (R & D Systems, PP-H6939-00), anti-IgG (CST, #3900) and anti-HIC1 (Sigma, #SAB1412231). ..

    Article Title: MicroRNAs Overcome Cell Fate Barrier by Reducing EZH2-Controlled REST Stability during Neuronal Conversion of Human Adult Fibroblasts
    Article Snippet: Crude nuclei were isolated in SONIC buffer (50mM HEPES (pH7.6), 140mM NaCl, 1mM EDTA, 1% Triton X-100, 0.1% Na-deoxyxhilate and 0.1% SDS) and were sonicated with probe-based sonicator. .. The sheared chromatin samples were immunoprecipitated overnight at 4 °C under gentle rotation with anti-IgG (CST, #2729), or anti-REST (Bethyl, A300-539) antibody, and then add ChIP-grade protein G magnetic beads to each IP reaction and incubate for 2h at 4 °C with rotati on. ..

    Incubation:

    Article Title: Progranulin Gene Therapy Improves Lysosomal Dysfunction and Microglial Pathology Associated with Frontotemporal Dementia and Neuronal Ceroid Lipofuscinosis
    Article Snippet: The following day, the membranes were incubated in an IR-Dye-conjugated anti-mouse antibody (1:20 000; LI-COR Biotechnology). .. With each round of testing, one membrane was incubated with a sheep anti-mouse progranulin antibody (#AF2557; R & D Systems) or rabbit anti-GFP antibody (#2956; Cell Signaling Technology) as a positive control. .. The progranulin antibody was detected by incubation with a biotinylated anti-sheep antibody (Vector Laboratories), followed by IR-Dye 800-conjugated streptavidin (LI-COR Biotechnology), and the GFP antibody was detected with an IR-Dye 680LT-conjugated anti-rabbit secondary antibody (LI-COR Biotechnology).

    Positive Control:

    Article Title: Progranulin Gene Therapy Improves Lysosomal Dysfunction and Microglial Pathology Associated with Frontotemporal Dementia and Neuronal Ceroid Lipofuscinosis
    Article Snippet: The following day, the membranes were incubated in an IR-Dye-conjugated anti-mouse antibody (1:20 000; LI-COR Biotechnology). .. With each round of testing, one membrane was incubated with a sheep anti-mouse progranulin antibody (#AF2557; R & D Systems) or rabbit anti-GFP antibody (#2956; Cell Signaling Technology) as a positive control. .. The progranulin antibody was detected by incubation with a biotinylated anti-sheep antibody (Vector Laboratories), followed by IR-Dye 800-conjugated streptavidin (LI-COR Biotechnology), and the GFP antibody was detected with an IR-Dye 680LT-conjugated anti-rabbit secondary antibody (LI-COR Biotechnology).

    Negative Control:

    Article Title: Novel Regulatory Roles of Wnt1 in Infection-Associated Colorectal Cancer
    Article Snippet: .. To evaluate ubiquitination of Wnt1, we did immunoprecipitation using anti-ubiquitin, anti-Wnt1, or anti-IgG (negative control) antibodies, respectively. ..

    Magnetic Beads:

    Article Title: MicroRNAs Overcome Cell Fate Barrier by Reducing EZH2-Controlled REST Stability during Neuronal Conversion of Human Adult Fibroblasts
    Article Snippet: Crude nuclei were isolated in SONIC buffer (50mM HEPES (pH7.6), 140mM NaCl, 1mM EDTA, 1% Triton X-100, 0.1% Na-deoxyxhilate and 0.1% SDS) and were sonicated with probe-based sonicator. .. The sheared chromatin samples were immunoprecipitated overnight at 4 °C under gentle rotation with anti-IgG (CST, #2729), or anti-REST (Bethyl, A300-539) antibody, and then add ChIP-grade protein G magnetic beads to each IP reaction and incubate for 2h at 4 °C with rotati on. ..

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  • 99
    Cell Signaling Technology Inc rabbit anti ha monoclonal antibody
    LET-413 preferentially associated with the active RAB-5 and the inactive RAB-10. (A) let-413 open reading frames encoding let-413a , let-413b , and let-413c isoforms. Red arrowheads above let-413c indicate three single guide RNAs target locations in let-413(ycx23) CRISPR/Cas9 intestine somatic mutants. (B) LET-413c contains N-terminal LRRs and a C-terminal PDZ domain; amino acid numbers are indicated. (C) Glutathione beads loaded with GST and GST-LET-413 were incubated with in vitro–expressed HA-tagged RAB-5(Q78L), RAB-10(Q68L), RAB-8(Q67L), RAB-11(Q70L), RAB-35(Q69L), and ARF-6(Q67L) and inactive RAB-5(S33N), RAB-10(T23N), RAB-8(T22N), RAB-11(S25N), RAB-35(S24N), and ARF-6(T27N). Eluted proteins were separated on SDS-PAGE gel and analyzed by Western blotting using <t>anti–HA</t> antibody. Input lanes contain in vitro–expressed HA-tagged proteins in the binding assays (10%). (D) The binding affinity of GST-LET-413 was fourfold higher for RAB-5(Q78L) than for RAB-5(S33N). The SEMs from three independent experiments are shown. Asterisks indicate significant differences (**, P
    Rabbit Anti Ha Monoclonal Antibody, supplied by Cell Signaling Technology Inc, used in various techniques. Bioz Stars score: 99/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/rabbit anti ha monoclonal antibody/product/Cell Signaling Technology Inc
    Average 99 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    rabbit anti ha monoclonal antibody - by Bioz Stars, 2021-07
    99/100 stars
      Buy from Supplier

    95
    Cell Signaling Technology Inc rabbit anti ha tag mab
    Ccr1l1 encodes a transmembrane protein that traffics to the plasma membrane and contains an extracellular N terminus and an intracellular C terminus. A , schematic representation of the pNT1 and pNT2 constructs cloned into a pcDNA3.1 plasmid for the expression of Ccr1l1 with a C-terminal or N-terminal <t>HA</t> <t>tag,</t> respectively. Ccr1l1 ORF is indicated by a black box , whereas the HA tag (HA) and kozak sequence (K) are shown in white boxes . The position of the ATG initiation codon is indicated above each graph. The expression of these constructs was analyzed by fluorescence microscopy ( B ) and FACS ( C and D ) in HEK293-transfected cells. B , fluorescence microscopy images of the <t>anti-HA</t> ( green ) and DAPI ( blue ) staining of HEK293 cells transfected with the plasmids indicated on the left of each row with (+Triton) or without (–Triton) permeabilization, as indicated above each column (400× magnification, scale bar = 20 μm). C , FACS histograms of the anti-HA staining of permeabilized or nonpermeabilized (as indicated above each graph) HEK293 cells transfected with the plasmids indicated in the insets of the right graph. These results are quantified in panel D . Bars represent the mean ± SD median fluorescence intensity (MFI) of three independent transfections from one experiment representative of three independent experiments. p values from a one-way ANOVA with Tukey correction for multiple comparisons are indicated.
    Rabbit Anti Ha Tag Mab, supplied by Cell Signaling Technology Inc, used in various techniques. Bioz Stars score: 95/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/rabbit anti ha tag mab/product/Cell Signaling Technology Inc
    Average 95 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    rabbit anti ha tag mab - by Bioz Stars, 2021-07
    95/100 stars
      Buy from Supplier

    Image Search Results


    LET-413 preferentially associated with the active RAB-5 and the inactive RAB-10. (A) let-413 open reading frames encoding let-413a , let-413b , and let-413c isoforms. Red arrowheads above let-413c indicate three single guide RNAs target locations in let-413(ycx23) CRISPR/Cas9 intestine somatic mutants. (B) LET-413c contains N-terminal LRRs and a C-terminal PDZ domain; amino acid numbers are indicated. (C) Glutathione beads loaded with GST and GST-LET-413 were incubated with in vitro–expressed HA-tagged RAB-5(Q78L), RAB-10(Q68L), RAB-8(Q67L), RAB-11(Q70L), RAB-35(Q69L), and ARF-6(Q67L) and inactive RAB-5(S33N), RAB-10(T23N), RAB-8(T22N), RAB-11(S25N), RAB-35(S24N), and ARF-6(T27N). Eluted proteins were separated on SDS-PAGE gel and analyzed by Western blotting using anti–HA antibody. Input lanes contain in vitro–expressed HA-tagged proteins in the binding assays (10%). (D) The binding affinity of GST-LET-413 was fourfold higher for RAB-5(Q78L) than for RAB-5(S33N). The SEMs from three independent experiments are shown. Asterisks indicate significant differences (**, P

    Journal: The Journal of Cell Biology

    Article Title: LET-413/Erbin acts as a RAB-5 effector to promote RAB-10 activation during endocytic recycling

    doi: 10.1083/jcb.201705136

    Figure Lengend Snippet: LET-413 preferentially associated with the active RAB-5 and the inactive RAB-10. (A) let-413 open reading frames encoding let-413a , let-413b , and let-413c isoforms. Red arrowheads above let-413c indicate three single guide RNAs target locations in let-413(ycx23) CRISPR/Cas9 intestine somatic mutants. (B) LET-413c contains N-terminal LRRs and a C-terminal PDZ domain; amino acid numbers are indicated. (C) Glutathione beads loaded with GST and GST-LET-413 were incubated with in vitro–expressed HA-tagged RAB-5(Q78L), RAB-10(Q68L), RAB-8(Q67L), RAB-11(Q70L), RAB-35(Q69L), and ARF-6(Q67L) and inactive RAB-5(S33N), RAB-10(T23N), RAB-8(T22N), RAB-11(S25N), RAB-35(S24N), and ARF-6(T27N). Eluted proteins were separated on SDS-PAGE gel and analyzed by Western blotting using anti–HA antibody. Input lanes contain in vitro–expressed HA-tagged proteins in the binding assays (10%). (D) The binding affinity of GST-LET-413 was fourfold higher for RAB-5(Q78L) than for RAB-5(S33N). The SEMs from three independent experiments are shown. Asterisks indicate significant differences (**, P

    Article Snippet: The antibodies used in this study were rabbit anti–actin polyclonal antibody (sc-1616-R; Santa Cruz Biotechnology), rabbit anti–HA monoclonal antibody (C29F4; Cell Signaling Technology), rabbit anti–GST monoclonal antibody (91G1; Cell Signaling Technology), rabbit anti–GFP polyclonal antibody ChIP grade (ab290; Abcam), rabbit anti–Erbin polyclonal antibody (ab55930; Abcam), rabbit anti–Rab5 monoclonal antibody (3547; Cell Signaling Technology), and anti–rabbit IgG-Alexa Fluor 594 conjugate (8889; Cell Signaling Technology).

    Techniques: CRISPR, Incubation, In Vitro, SDS Page, Western Blot, Binding Assay

    LET-413 preferentially associated with the active RAB-5 and the inactive RAB-10. (A) let-413 open reading frames encoding let-413a , let-413b , and let-413c isoforms. Red arrowheads above let-413c indicate three single guide RNAs target locations in let-413(ycx23) CRISPR/Cas9 intestine somatic mutants. (B) LET-413c contains N-terminal LRRs and a C-terminal PDZ domain; amino acid numbers are indicated. (C) Glutathione beads loaded with GST and GST-LET-413 were incubated with in vitro–expressed HA-tagged RAB-5(Q78L), RAB-10(Q68L), RAB-8(Q67L), RAB-11(Q70L), RAB-35(Q69L), and ARF-6(Q67L) and inactive RAB-5(S33N), RAB-10(T23N), RAB-8(T22N), RAB-11(S25N), RAB-35(S24N), and ARF-6(T27N). Eluted proteins were separated on SDS-PAGE gel and analyzed by Western blotting using anti–HA antibody. Input lanes contain in vitro–expressed HA-tagged proteins in the binding assays (10%). (D) The binding affinity of GST-LET-413 was fourfold higher for RAB-5(Q78L) than for RAB-5(S33N). The SEMs from three independent experiments are shown. Asterisks indicate significant differences (**, P

    Journal: The Journal of Cell Biology

    Article Title: LET-413/Erbin acts as a RAB-5 effector to promote RAB-10 activation during endocytic recycling

    doi: 10.1083/jcb.201705136

    Figure Lengend Snippet: LET-413 preferentially associated with the active RAB-5 and the inactive RAB-10. (A) let-413 open reading frames encoding let-413a , let-413b , and let-413c isoforms. Red arrowheads above let-413c indicate three single guide RNAs target locations in let-413(ycx23) CRISPR/Cas9 intestine somatic mutants. (B) LET-413c contains N-terminal LRRs and a C-terminal PDZ domain; amino acid numbers are indicated. (C) Glutathione beads loaded with GST and GST-LET-413 were incubated with in vitro–expressed HA-tagged RAB-5(Q78L), RAB-10(Q68L), RAB-8(Q67L), RAB-11(Q70L), RAB-35(Q69L), and ARF-6(Q67L) and inactive RAB-5(S33N), RAB-10(T23N), RAB-8(T22N), RAB-11(S25N), RAB-35(S24N), and ARF-6(T27N). Eluted proteins were separated on SDS-PAGE gel and analyzed by Western blotting using anti–HA antibody. Input lanes contain in vitro–expressed HA-tagged proteins in the binding assays (10%). (D) The binding affinity of GST-LET-413 was fourfold higher for RAB-5(Q78L) than for RAB-5(S33N). The SEMs from three independent experiments are shown. Asterisks indicate significant differences (**, P

    Article Snippet: Antibodies The antibodies used in this study were rabbit anti–actin polyclonal antibody (sc-1616-R; Santa Cruz Biotechnology), rabbit anti–HA monoclonal antibody (C29F4; Cell Signaling Technology), rabbit anti–GST monoclonal antibody (91G1; Cell Signaling Technology), rabbit anti–GFP polyclonal antibody ChIP grade (ab290; Abcam), rabbit anti–Erbin polyclonal antibody (ab55930; Abcam), rabbit anti–Rab5 monoclonal antibody (3547; Cell Signaling Technology), and anti–rabbit IgG-Alexa Fluor 594 conjugate (8889; Cell Signaling Technology).

    Techniques: CRISPR, Incubation, In Vitro, SDS Page, Western Blot, Binding Assay

    SAC-1 interacts with the active and inactive mutant forms of ARF-6, and the residence of SAC-1 in basolateral puncta requires ARF-6. (A) Western blot showing GST pull-down with in vitro–translated HA-tagged proteins. Glutathione beads loaded with GST and GST-SAC-1 were incubated with in vitro–expressed HA-tagged RAB-5(Q78L), RAB-7(Q68L), RAB-8(Q67L), RAB-10(Q68L), RAB-11(Q70L), ARF-6(Q67L), and ARF-6(T27N). Eluted proteins were separated on the SDS-PAGE gel and analyzed by Western blotting using anti–HA antibody. Input lanes contain in vitro–expressed HA-tagged proteins in the binding assays (5%). (B and B′) SAC-1 colocalized well with ARF-6–, RAB-5–, or RAB-10–labeled endosomes. In addition, SAC-1 overlaps significantly with the TGN marker AMAN-2 and the ER marker SP12. Nevertheless, SAC-1 displayed little colocalization with the late endosome marker RAB-7. Arrowheads indicate positive overlap. Pearson’s correlation coefficients for GFP and mCherry signals were calculated ( n = 6 animals). (C and C′) Compared with wild-type animals, the labeling of GFP-SAC-1 in basolateral puncta was significantly reduced in the absence of ARF-6 (top focal plane). Transgenic expression of ARF-6(Q67L)-mCherry rescued the puncta labeling GFP-SAC-1, and the transgenic expression of ARF-6(T27N)-mCherry failed to restore the endosomal localization of GFP-SAC-1. Asterisks in the panels indicate intestinal lumen. Error bars represent SEM ( n = 18 each), and asterisks indicate the significant differences in the one-tailed Student’s t test (ns, no significance; **, P

    Journal: The Journal of Cell Biology

    Article Title: SAC-1 ensures epithelial endocytic recycling by restricting ARF-6 activity

    doi: 10.1083/jcb.201711065

    Figure Lengend Snippet: SAC-1 interacts with the active and inactive mutant forms of ARF-6, and the residence of SAC-1 in basolateral puncta requires ARF-6. (A) Western blot showing GST pull-down with in vitro–translated HA-tagged proteins. Glutathione beads loaded with GST and GST-SAC-1 were incubated with in vitro–expressed HA-tagged RAB-5(Q78L), RAB-7(Q68L), RAB-8(Q67L), RAB-10(Q68L), RAB-11(Q70L), ARF-6(Q67L), and ARF-6(T27N). Eluted proteins were separated on the SDS-PAGE gel and analyzed by Western blotting using anti–HA antibody. Input lanes contain in vitro–expressed HA-tagged proteins in the binding assays (5%). (B and B′) SAC-1 colocalized well with ARF-6–, RAB-5–, or RAB-10–labeled endosomes. In addition, SAC-1 overlaps significantly with the TGN marker AMAN-2 and the ER marker SP12. Nevertheless, SAC-1 displayed little colocalization with the late endosome marker RAB-7. Arrowheads indicate positive overlap. Pearson’s correlation coefficients for GFP and mCherry signals were calculated ( n = 6 animals). (C and C′) Compared with wild-type animals, the labeling of GFP-SAC-1 in basolateral puncta was significantly reduced in the absence of ARF-6 (top focal plane). Transgenic expression of ARF-6(Q67L)-mCherry rescued the puncta labeling GFP-SAC-1, and the transgenic expression of ARF-6(T27N)-mCherry failed to restore the endosomal localization of GFP-SAC-1. Asterisks in the panels indicate intestinal lumen. Error bars represent SEM ( n = 18 each), and asterisks indicate the significant differences in the one-tailed Student’s t test (ns, no significance; **, P

    Article Snippet: Antibodies The antibodies used in the current study are listed below: rabbit anti–actin polyclonal antibody (sc-1616-R; Santa Cruz Biotechnologies), rabbit anti–HA monoclonal antibody (C29F4; Cell Signaling Technology), rabbit anti–GST monoclonal antibody (91G1; Cell Signaling Technology), rabbit anti–mCherry polyclonal antibody (ab167453; Abcam), rabbit anti–GFP polyclonal antibody–chromatin immunoprecipitation grade (ab290; Abcam), mouse anti–P(4,5)P2 monoclonal antibody (Z-P045; Echelon Biosciences), and mouse anti–PIPn monoclonal antibody (Z-P999; Echelon Biosciences).

    Techniques: Mutagenesis, Western Blot, In Vitro, Incubation, SDS Page, Binding Assay, Labeling, Marker, Transgenic Assay, Expressing, One-tailed Test

    Ccr1l1 encodes a transmembrane protein that traffics to the plasma membrane and contains an extracellular N terminus and an intracellular C terminus. A , schematic representation of the pNT1 and pNT2 constructs cloned into a pcDNA3.1 plasmid for the expression of Ccr1l1 with a C-terminal or N-terminal HA tag, respectively. Ccr1l1 ORF is indicated by a black box , whereas the HA tag (HA) and kozak sequence (K) are shown in white boxes . The position of the ATG initiation codon is indicated above each graph. The expression of these constructs was analyzed by fluorescence microscopy ( B ) and FACS ( C and D ) in HEK293-transfected cells. B , fluorescence microscopy images of the anti-HA ( green ) and DAPI ( blue ) staining of HEK293 cells transfected with the plasmids indicated on the left of each row with (+Triton) or without (–Triton) permeabilization, as indicated above each column (400× magnification, scale bar = 20 μm). C , FACS histograms of the anti-HA staining of permeabilized or nonpermeabilized (as indicated above each graph) HEK293 cells transfected with the plasmids indicated in the insets of the right graph. These results are quantified in panel D . Bars represent the mean ± SD median fluorescence intensity (MFI) of three independent transfections from one experiment representative of three independent experiments. p values from a one-way ANOVA with Tukey correction for multiple comparisons are indicated.

    Journal: The Journal of Biological Chemistry

    Article Title: Structural and functional analysis of Ccr1l1, a Rodentia-restricted eosinophil-selective chemokine receptor homologue

    doi: 10.1016/j.jbc.2021.100373

    Figure Lengend Snippet: Ccr1l1 encodes a transmembrane protein that traffics to the plasma membrane and contains an extracellular N terminus and an intracellular C terminus. A , schematic representation of the pNT1 and pNT2 constructs cloned into a pcDNA3.1 plasmid for the expression of Ccr1l1 with a C-terminal or N-terminal HA tag, respectively. Ccr1l1 ORF is indicated by a black box , whereas the HA tag (HA) and kozak sequence (K) are shown in white boxes . The position of the ATG initiation codon is indicated above each graph. The expression of these constructs was analyzed by fluorescence microscopy ( B ) and FACS ( C and D ) in HEK293-transfected cells. B , fluorescence microscopy images of the anti-HA ( green ) and DAPI ( blue ) staining of HEK293 cells transfected with the plasmids indicated on the left of each row with (+Triton) or without (–Triton) permeabilization, as indicated above each column (400× magnification, scale bar = 20 μm). C , FACS histograms of the anti-HA staining of permeabilized or nonpermeabilized (as indicated above each graph) HEK293 cells transfected with the plasmids indicated in the insets of the right graph. These results are quantified in panel D . Bars represent the mean ± SD median fluorescence intensity (MFI) of three independent transfections from one experiment representative of three independent experiments. p values from a one-way ANOVA with Tukey correction for multiple comparisons are indicated.

    Article Snippet: Then, 400,000 cells were stained with Zombie Violet viability dye (Biolegend) for 10 min at room temperature and a rabbit anti-HA tag mAb (Cell Signaling Technology, Danvers, MA) on ice for 20 min followed by an anti-rabbit F(ab′)2 fragment conjugated with AlexaFluor 488 (Cell Signaling Technology).

    Techniques: Construct, Clone Assay, Plasmid Preparation, Expressing, Sequencing, Fluorescence, Microscopy, FACS, Transfection, Staining