SP1

SP1 CUTANA™ CUT&RUN Antibody

$495.00
SKU: 13-2024
Pack Size: 100 µL
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  • Type:  Polyclonal
  • Host:  Rabbit
  • Applications:  CUT&RUN, IHC, IP, WB
  • Reactivity:  Human, Mouse
  • Format:  Antigen affinity-purified
  • Target Size:  81 kDa

Description

This antibody meets EpiCypher’s "CUTANA Compatible" criteria for performance in Cleavage Under Targets and Release Using Nuclease (CUT&RUN) and/or Cleavage Under Targets and Tagmentation (CUT&Tag) approaches to genomic mapping. Every lot of a CUTANA Compatible antibody is tested in the indicated approach using EpiCypher optimized protocols and determined to yield peaks that show a genomic distribution pattern consistent with reported function(s) of the target protein. SP1 is the original member of the Sp1-like family of zinc-finger transcription factors. It binds GC-rich motifs with high affinity [1] and is involved in expressing genes related to cell growth, cell-cycle progression, survival, and tumorigenesis [2]. SP1 antibody produces CUT&RUN peaks above background primarily in intronic and promoter regions (Figure 1) that overlap with known SP1 DNA-binding motifs (Figure 2).

Validation Data

CUT&RUN was performed on 500k HeLa cells with 0.5 µg of either SP1 or IgG negative control (EpiCypher 13-0042) antibodies using the CUTANA™ ChIC/CUT&RUN Kit v2.0 (EpiCypher 14-1048). Library preparation was performed with 5 ng of DNA (or the total amount recovered if less than 5 ng) using the CUTANA™ CUT&RUN Library Prep Kit (EpiCypher 14-1001/14-1002). Both kit protocols were adapted for high throughput Tecan liquid handling. Libraries were run on an Illumina NextSeq2000 with paired-end sequencing (2x50 bp). Sample sequencing depth was 5.0 million reads (IgG) and 5.3 million reads (SP1). Data were aligned to the hg19 genome using Bowtie2. Data were filtered to remove duplicates, multi-aligned reads, and ENCODE DAC Exclusion List regions.

13-2024-sp1-peaks (Click to enlarge)

Figure 1: SP1 peaks in CUT&RUN
CUT&RUN was performed as described above. Peaks were called with MACS2. Heatmaps show SP1 peaks relative to IgG negative control antibody in aligned rows ranked by intensity (top to bottom) and colored such that red indicates high localized enrichment and blue denotes background signal (left). The number of peaks that fall into distinct classes of functionally annotated genomic regions are shown (right).

13-2024-binding-motif-analysis (Click to enlarge)

Figure 2: SP1 transcription factor binding motif analysis in CUT&RUN
Sp1(Zf)/Promoter/Homer consensus motif, represented as a sequence logo position weight matrix, was the top called motif significantly enriched under SP1 CUT&RUN peaks (left). The number of SP1 peaks containing the consensus motif from the left panel is represented by a Venn Diagram (right).

13-2024-representative-browser-tracks (Click to enlarge)

Figure 3: SP1 CUT&RUN representative browser tracks
CUT&RUN was performed as described above. Gene browser shots were generated using the Integrative Genomics Viewer (IGV, Broad Institute). Three representative loci of the top called peaks are shown.

13-2024-immunohistochemistry-data (Click to enlarge)

Figure 4: Immunohistochemistry data
FFPE sections of human breast carcinoma (left) and mouse renal cell carcinoma (right) using SP1 antibody at a dilution of 1:5,000.

13-2024-immunoprecipitation-data (Click to enlarge)

Figure 5: Immunoprecipitation data
EpiCypher SP1 antibody (6 µg) was used to immunoprecipitate whole cell lysates (0.5 - 1 mg, 20% of IP loaded) isolated from HeLa cells using NETN lysis buffer. A negative control IgG antibody and positive control antibodies targeting SP1 (Bethyl Laboratories) were also used to demonstrate specificity of the IP. EpiCypher 13-2024 and Bethyl A300-134A-5 target the same epitope, while Bethyl A300-944A-1 targets a different epitope (between amino acids 735 and 785). For blotting immunoprecipitates, EpiCypher SP1 antibody was used at a 1:1,000 dilution.

13-2024-western-blot-data (Click to enlarge)

Figure 6: Western blot data
Western analysis of SP1 in whole cell extracts from HeLa, HEK293T, and mouse Renca cells. Fifty micrograms of lysate was resolved via SDS-PAGE and detected with 1:10,000 dilution of SP1 antibody.

Technical Information

Immunogen
Between amino acids 750 and 785
Storage
Stable for 1 year at 4°C from date of receipt
Formulation
Antigen affinity-purified antibody in Tris-citrate/phosphate buffer pH 7-8, 0.09% sodium azide

Recommended Dilution

CUT&RUN:
0.5 µg per reaction
Immunohistochemistry (IHC):
1:2,000 - 1:10,000. Epitope retrieval with citrate buffer pH 6.0 is recommended for FFPE tissue sections
Immunoprecipitation (IP):
2 - 10 µg/mg lysate
Western Blot (WB):
1:2,000 - 1:10,000

Gene & Protein Information

UniProt ID
P08047
Gene Name
SP1
Protein Name
Transcription factor SP1
Target Size
81 kDa
Alternate Names
specificity protein 1, transcription factor SP1, TSFP1

References

Background References:
[1] Raiber et al. Nucleic Acids Res (2012). PMID: 22021377
[2] Spengler et al. Cell Cycle (2008). PMID: 18239466

Documents & Resources

Technical Datasheet

Additional Info

This antibody is provided for commercial sale under license from Bethyl Laboratories, Inc.

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