Anti-SAP62 antibody [4G8] (ab77800)

Overview

• Product nameAnti-SAP62 antibody [4G8]See all SAP62 primary antibodies ...
• Description
  Mouse monoclonal [4G8] to SAP62
• Tested applicationsIHC-P, WB, ICC/IF, IP, Flow Cyt more details
• Species reactivity
  Reacts with: Rat, Human, Monkey
  
• Immunogen

  Nuclear protein isolated from rat liver nuclei.

• EpitopeC-terminal, amino acids 212-464.
• Positive controlThis antibody gave a positive signal in the following whole cell lysates: HeLa, HepG2

Properties

• FormLiquid
• Storage instructionsStore at +4°C short term (1-2 weeks). Aliquot and store at -20°C or -80°C. Avoid repeated freeze / thaw cycles.
• Storage bufferPreservative: 0.02% Sodium Azide
  Constituents: PBS, pH 7.4
• Concentration information loading...
• PurityIgG fraction
• Clonality Monoclonal
• Clone number4G8
• MyelomaSp2/0
• IsotypeIgG1
• Research Areas
  • Epigenetics and Nuclear Signaling
  • DNA / RNA
  • RNA Processing
  • Splicing

Applications

Target

• FunctionSubunit of the splicing factor SF3A required for 'A' complex assembly formed by the stable binding of U2 snRNP to the branchpoint sequence (BPS) in pre-mRNA. Sequence independent binding of SF3A/SF3B complex upstream of the branch site is essential, it may anchor U2 snRNP to the pre-mRNA. May also be involved in the assembly of the 'E' complex.
• Sequence similaritiesBelongs to the SF3A2 family.
  Contains 1 matrin-type zinc finger.
• Cellular localizationNucleus.

Target information above from: UniProt accession Q15428 The UniProt Consortium
The Universal Protein Resource (UniProt) in 2010
Nucleic Acids Res. 38:D142-D148 (2010) .

Information by UniProt
• Database links
  • Entrez Gene: 8175 Human
  • Entrez Gene: 8175 Human
  • Entrez Gene: 299620 Rat
  • Omim: 600796 Human
  • SwissProt: Q15428 Human
  • SwissProt: Q15428 Human
  • SwissProt: Q6AXT8 Rat
  • Unigene: 115232 Human

  • Unigene: 102569 Rat

  see all

• Alternative names
  pre-mRNA splicing factor SF3A, subunit 2 antibodyPRP11 antibodyPRPF11 antibody

  SAP 62 antibodySf3a2 antibodySF3A2_HUMAN antibodySF3a66 antibodyspliceosome associated protein 62 antibodySpliceosome-associated protein 62 antibodysplicing factor 3a subunit 2 antibodysplicing factor 3a, subunit 2, 66kDa antibody

  see all

Anti-SAP62 antibody [4G8] images

• 

  Immunocytochemistry/ Immunofluorescence - SAP62 antibody [4G8] (ab77800)
  ICC/IF image of ab77800 stained Hek293 cells. The cells were 4% PFA fixed (10 min) and then incubated in 1%BSA / 10% normal gost serum / 0.3M glycine in 0.1% PBS-Tween for 1h to permeabilise the cells and block non-specific protein-protein interactions. The cells were then incubated with the antibody (ab77800, 5µg/ml) overnight at +4°C. The secondary antibody (green) was Alexa Fluor® 488 goat anti-mouse IgG (H+L) used at a 1/1000 dilution for 1h. Alexa Fluor® 594 WGA was used to label plasma membranes (red) at a 1/200 dilution for 1h. DAPI was used to stain the cell nuclei (blue) at a concentration of 1.43µM. This antibody also gave a positive result in 4% PFA fixed (10 min) HeLa, HepG2 and MCF7 cells at 5µg/ml, and in 100% methanol fixed (5 min) HeLa, Hek293, HepG2 and MCF7 cells at 5µg/ml.
• 

  Western blot - SAP62 antibody [4G8] (ab77800)
  All lanes : Anti-SAP62 antibody [4G8] (ab77800) at 1 µg/ml
  
  Lane 1 : HeLa (Human epithelial carcinoma cell line) Nuclear Lysate
  Lane 2 : HepG2 (Human hepatocellular liver carcinoma cell line) Nuclear Lysate (ab14660)
  
  Lysates/proteins at 20 µg per lane.
  
  Secondary
  Goat polyclonal to Mouse IgG - H&L - Pre-Adsorbed (HRP) at 1/3000 dilution
  developed using the ECL technique
  
  Performed under reducing conditions.
  
  Predicted band size : 52 kDa
  Observed band size : 60 kDa (why is the actual band size different from the predicted?)
  
  
  Exposure time : 3 minutes
• 

  Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - SAP62 antibody [4G8] (ab77800)
  IHC image of 77800 staining in human normal skin formalin fixed paraffin embedded tissue section, performed on a Leica Bond^TM system using the standard protocol F. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH6, epitope retrieval solution 1) for 20 mins. The section was then incubated with ab77800, 1µg/ml, for 15 mins at room temperature and detected using an HRP conjugated compact polymer system. DAB was used as the chromogen. The section was then counterstained with haematoxylin and mounted with DPX.
• 

  Flow Cytometry-Anti-SAP62 antibody [4G8](ab77800)
  Overlay histogram showing HeLa cells stained with ab77800 (red line). The cells were fixed with 80% methanol (5 min) and then permeabilized with 0.1% PBS-Tween for 20 min. The cells were then incubated in 1x PBS / 10% normal goat serum / 0.3M glycine to block non-specific protein-protein interactions followed by the antibody (ab77800, 1µg/1x10⁶ cells) for 30 min at 22ºC. The secondary antibody used was DyLight® 488 goat anti-mouse IgG (H+L) (ab96879) at 1/500 dilution for 30 min at 22ºC. Isotype control antibody (black line) was mouse IgG1 [ICIGG1] (ab91353, 2µg/1x10⁶ cells) used under the same conditions. Acquisition of >5,000 events was performed. This antibody gave a positive signal in HeLa cells fixed with 4% paraformaldehyde (10 min)/permeabilized with 0.1% PBS-Tween for 20 min used under the same conditions.