Antibody data
- Antibody Data
- Antigen structure
- References [1]
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- Validations
- Western blot [2]
- Immunohistochemistry [2]
- Other assay [3]
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- Product number
- PA5-38024 - Provider product page
- Provider
- Invitrogen Antibodies
- Product name
- WAPL Polyclonal Antibody
- Antibody type
- Polyclonal
- Antigen
- Synthetic peptide
- Description
- A recommended positive control is A-20 cell lysate.
- Concentration
- 1 mg/mL
Submitted references Loop extrusion mediates physiological Igh locus contraction for RAG scanning.
Dai HQ, Hu H, Lou J, Ye AY, Ba Z, Zhang X, Zhang Y, Zhao L, Yoon HS, Chapdelaine-Williams AM, Kyritsis N, Chen H, Johnson K, Lin S, Conte A, Casellas R, Lee CS, Alt FW
Nature 2021 Feb;590(7845):338-343
Nature 2021 Feb;590(7845):338-343
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Supportive validation
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- Experimental details
- Western blot analysis of WAPL in A20 cell lysate using a WAPL polyclonal antibody (Product # PA5-38024) at a dilution of 1 µg/mL (A) and 2 µg/mL (B).
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- Experimental details
- Western Blot analysis of WAPL in A20 cell lysate with WAPL Polyclonal Antibody (Product # PA5-38024) at (A) 1 and (B) 2 µg/mL.
Supportive validation
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- Invitrogen Antibodies (provider)
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- Experimental details
- Immunofluorescence of WAPL in rat heart tissue with WAPL Polyclonal Antibody (Product # PA5-38024) at 20 µg/mL.
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- Invitrogen Antibodies (provider)
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- Experimental details
- Immunohistochemistry of WAPL in rat heart tissue with WAPL Polyclonal Antibody (Product # PA5-38024) at 5 µg/mL.
Supportive validation
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- Experimental details
- Figure. 4 | Wapl depletion activates IgH V H locus contraction and long-range V H utilization in G1-arrested v-Abl cells. a, Western blotting to detect Wapl protein levels in the primary #5 cycling v-Abl cells and the derived Wapl-degron #5-9 v-Abl cells before (day0) and after STI&IAA&Dox treatment (day4) and cultured primary pro-B cells. One of the two experiments is shown. b , Western blotting results to determine relative expression of Wapl protein levels in cycling and G1 arrested primary #5 v-Abl cells and cultured primary pro-B cells. Average values +- s.d. of Wapl protein levels are shown. For comparison, the intensity of the Wapl band in G1-arrested primary #5 v-Abl cells is set as 1.0. Average value is indicated at each bar. n , number of independent repeats. Indicated P values were calculated using unpaired two-tailed t -test. c, Average 3C-HTGTS signal counts +- s.e.m. across the four V H domains in cultured RAG1-deficient primary pro-B cells (top), G1-arrested RAG1-deficient v-Abl cells without (middle) or with Wapl depletion (bottom). n , number of independent experiments. For comparison, 15 representative major interaction peaks/clusters are shown as in Fig. 3b . d, Average utilization frequencies +- s.d. of all V H segments in RAG1-complemented, G1-arrested Wapl-degron v-Abl cells without (Untreated) or with (IAA&Dox) Wapl depletion are indicated. Average percentage +- s.d. of V H DJ H and DJ H rearrangements are shown. n , number of independent experiments. S
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- Extended Data Figure. 6 | Generation and characterization of RAG1-deficient Wapl-degron v-Abl cell lines. a , Scatter plots of average transcriptome-wide GRO-seq counts in G1-arrested v-Abl cells ( x axis , n=3) and primary pro-B cells ( y axis , n=4). n , number of independent experiments. Representative known requisite genes implicated in the cohesin-complex function for V(D)J recombination and chromatin interactions are highlighted by red circles and blue arrows. Representative known genes implicated in the DNA repair and B cell development were also analyzed to determine if there were any potential transcriptional defects in these essential genes for V(D)J recombination and none were found and highlighted. Analyses of scatter plots indicate that Wapl is expressed at significantly higher levels in G1-arrested v-Abl cells than in primary pro-B cells (Spearman's correlation coefficient (rho) and P values determined by two-sided Spearman's correlation test are presented). These transcription finding were confirmed by western blotting studies ( Fig. 4a ). b , Comparison of Wapl transcription levels by GRO-seq in primary pro-B cells and G1-arrested v-Abl cells from 4 and 3 independent repeats, respectively. Data are presented as average signal counts +- s.e.m. of GRO-seq. n , number of independent experiments. Although other genes upstream and downstream of the Wapl gene show altered transcription in primary pro-B cells compared to the v-Abl cells, their products thus far have
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- Extended Data Figure. 7 | Characterization of Wapl/ CTCF/ Rad21-binding in G1-arrested RAG1-deficient Wapl-degron v-Abl cells. a, Average signal counts +- s.e.m. of Wapl ChIP-seq across the entire IgH locus are plotted as indicated for G1-arrested RAG1-deficient v-Abl cells without (Untreated, blue) or with (IAA&Dox, red) Wapl depletion and cultured RAG1-deficient primary pro-B cells (green). n , number of independent experiments. Wapl ChIP-seq showed that IAA&Dox treatment leads to a depletion of chromatin-bound Wapl at IgH locus, which largely resembles that of primary pro-B cells at IgH locus. b , Three independent repeats of Wapl ChIP-seq signal within +-1.0 kb region across all peaks genome-wide called in G1-arrested RAG1-deficient v-Abl cells without (Untreated) or with (IAA&Dox) Wapl depletion. Top: Average enrichment. c , d , Average signal counts +- s.e.m. of CTCF ( c ) and Rad21 ( d ) ChIP-seq across the entire IgH locus are plotted as indicated for G1-arrested RAG1-deficient v-Abl cells without (Untreated, blue) or with (IAA&Dox, red) Wapl depletion, and cultured RAG1-deficient primary pro-B cells (green). n , number of independent experiments. Rad21 ChIP-seq showed that Wapl depletion in G1-arrested v-Abl cells influenced Rad21 (cohesin) redistribution across the IgH locus to give a pattern significantly similar to primary pro-B cells (Spearman's correlation r=0.84, P