GTPase Activating Protein Antibody (B4F8)

GTPase Activating Protein Monoclonal Antibody for Western blot, IF, IP

Synonyms:
GAP
Details
Host / Isotype: Mouse / IgG2a
Class: Monoclonal
Type: Antibody
Clone: B4F8
Tested Species Reactivity: Human (Hu), Mouse (Ms), Rat (Rt), Bovine (Bv), Non-human primate (Nhp)
Published Species Reactivity: Human (Hu)
Immunogen: Full length recombinant human GAP.
Ordering Information
Pierce GTPase Activating Protein Antibody (B4F8)
Product #
MA4-001
Size
100 µg
Price
$316.00
Purchase
Add GTPase Activating Protein Antibody (B4F8) to your cart.
Tested Applications Dilution *
Western Blot (WB) 10 µg/ml
Immunofluorescence (IF) 5 µg/ml
Immunoprecipitation (IP) 1-5 µg
Published Applications Dilution
Western Blot (WB) See publications below
Immunoprecipitation (IP) See publications below
* Suggested working dilutions are given as a guide only. It is recommended that the user titrates the product for use in their own experiment using appropriate negative and positive controls.
Form Information
Form: Lyophilized
Purification: Protein A
Preservative: no preservative
Storage Conditions: -20° C, Avoid Freeze/Thaw Cycles
Product Specific Information
MA4-001 detects GTPase activating protein (GAP) from human, mouse, rat, bovine and non-human primate tissues.

MA4-001 has been successfully used in Western blot, immunofluorescence and immunoprecipitation procedures. By Western blot, this antibody recognizes a single 120 kDa protein representing ras GAP from RS-2 cell lysate. Immunofluorescence staining of GAP in mouse fibroblast cells with MA4-001 results in diffuse cytoplasmic staining. Following the addition of PDGF, immunofluorescence staining shows that some GAP rapidly translocates to the plasma membrane. MA4-001 immunoprecipitates GAP that is complexed with the GAP related p190.

The MA4-001 antigen is full length recombinant human GAP. Epitope mapping studies suggest that this antibody binds a portion of GAP that contains the src homology regions SH2 & SH3.

Reconstitute with PBS.
General Information
GTPase Activating Protein (GAP) is capable of stimulating the hydrolysis of GTP by ras p21 proteins, though GAP has little effect on the oncogenic forms of ras. It is also known that several tyrosine kinases such as platelet derived growth factor receptor and epidermal growth factor receptor are involved in the tyrosine phosphorylation of GAP. It has therefore been suggested that GAP may provide a connection or link between growth factor receptors and the ras p21 family.
Product Images
  • GTPase Activating Protein Antibody (MA4-001) in WB
    MA4-001_Westernblot.jpg

    Western Blot with anti-GTPase Activating Protein Monoclonal Antibody [B4F8] (MA4-001) (MA4-001_Westernblot.jpg)

    Western Blot with anti-GTPase Activating Protein Monoclonal Antibody [B4F8] (MA4-001)

    Western blot analysis of GTPase Activating Protein was performed by loading 25 ug of Hela (Lane 1), A431 (Lane 2), and mouse brain cell lysates (Lane 3) and a molecular weight protein ladder onto an SDS polyacrylamide gel. Proteins were transferred to a PVDF membrane and blocked with a blocking buffer at 4ºC overnight. The membrane was probed with a GTPase Activating Protein monoclonal antibody (Product # MA4-001) at a dilution of 1:200 overnight at 4°C, washed in TBST, and probed with an HRP-conjugated secondary antibody for 1 hr at room temperature in the dark. Chemiluminescent detection was performed using Pierce ECL Plus Western Blotting Substrate (Product # 32132). Results show a band at 110 kDa in all three cell lines.
  • GTPase Activating Protein Antibody (MA4-001) in IF
    MA4-001_IF_Hela.jpg

    Immunofluorescence with anti-GTPase Activating Protein Monoclonal Antibody [B4F8] (MA4-001) (MA4-001_IF_Hela.jpg)

    Immunofluorescence with anti-GTPase Activating Protein Monoclonal Antibody [B4F8] (MA4-001)

    Immunofluorescent analysis of GTPase Activating Protein (green) showing positive staining in the cytoplasm of Hela cells (right) compared with a negative control in the absence of primary antibody (left). Formalin-fixed cells were permeabilized with 0.1% Triton X-100 in TBS for 5-10 minutes, blocked with 3% BSA-PBS for 30 minutes at room temperature and probed with a GTPase Activating Protein monoclonal antibody (Product # MA4-001) in 3% BSA-PBS at a dilution of 1:200 and incubated overnight at 4 ºC in a humidified chamber. Cells were washed with PBST and incubated with a DyLight 488-conjugated goat-anti-mouse IgG (H+L) secondary antibody in PBS at room temperature in the dark. F-actin (red) was stained with a flourescent red phalloidin and nuclei (blue) were stained with DAPI for 5-10 minutes in the dark. Images were taken at a magnification of 60x.
  • GTPase Activating Protein Antibody (MA4-001) in IF
    MA4-001_IF_C2C12.jpg

    Immunofluorescence with anti-GTPase Activating Protein Monoclonal Antibody [B4F8] (MA4-001) (MA4-001_IF_C2C12.jpg)

    Immunofluorescence with anti-GTPase Activating Protein Monoclonal Antibody [B4F8] (MA4-001)

    Immunofluorescent analysis of GTPase Activating Protein (green) showing positive staining in the cytoplasm of C2C12 cells (right) compared with a negative control in the absence of primary antibody (left). Formalin-fixed cells were permeabilized with 0.1% Triton X-100 in TBS for 5-10 minutes, blocked with 3% BSA-PBS for 30 minutes at room temperature and probed with a GTPase Activating Protein monoclonal antibody (Product # MA4-001) in 3% BSA-PBS at a dilution of 1:200 and incubated overnight at 4 ºC in a humidified chamber. Cells were washed with PBST and incubated with a DyLight 488-conjugated goat-anti-mouse IgG (H+L) secondary antibody in PBS at room temperature in the dark. F-actin (red) was stained with a flourescent red phalloidin and nuclei (blue) were stained with DAPI for 5-10 minutes in the dark. Images were taken at a magnification of 60x.
  • GTPase Activating Protein Antibody (MA4-001) in IF
    MA4-001_IF_A431.jpg

    Immunofluorescence with anti-GTPase Activating Protein Monoclonal Antibody [B4F8] (MA4-001) (MA4-001_IF_A431.jpg)

    Immunofluorescence with anti-GTPase Activating Protein Monoclonal Antibody [B4F8] (MA4-001)

    Immunofluorescent analysis of GTPase Activating Protein (green) showing positive staining in the cytoplasm of A431 cells (right) compared with a negative control in the absence of primary antibody (left). Formalin-fixed cells were permeabilized with 0.1% Triton X-100 in TBS for 5-10 minutes, blocked with 3% BSA-PBS for 30 minutes at room temperature and probed with a GTPase Activating Protein monoclonal antibody (Product # MA4-001) in 3% BSA-PBS at a dilution of 1:200 and incubated overnight at 4 ºC in a humidified chamber. Cells were washed with PBST and incubated with a DyLight 488-conjugated goat-anti-mouse IgG (H+L) secondary antibody in PBS at room temperature in the dark. F-actin (red) was stained with a flourescent red phalloidin and nuclei (blue) were stained with DAPI for 5-10 minutes in the dark. Images were taken at a magnification of 60x.
Publications:
Western Blot
  Species / Dilution Summary
  Hu / Not Cited
MA4-001 was used in immunoprecipitation and western blot to clone the GAP-associated protein p190 and to investigate its functional properties

Cell. 1992 May 1;69(3):539-49.
"Molecular cloning of cDNAs encoding the GAP-associated protein p190: implications for a signaling pathway from ras to the nucleus."
Author(s): Settleman J, Narasimhan V, Foster LC, Weinberg RA
Number of Citations: 75
(See PubMed article )
Immunoprecipitation
  Species / Dilution Summary
  Hu / Not Cited
MA4-001 was used in immunoprecipitation and western blot to clone the GAP-associated protein p190 and to investigate its functional properties

Cell. 1992 May 1;69(3):539-49.
"Molecular cloning of cDNAs encoding the GAP-associated protein p190: implications for a signaling pathway from ras to the nucleus."
Author(s): Settleman J, Narasimhan V, Foster LC, Weinberg RA
Number of Citations: 75
(See PubMed article )
(This product is for In Vitro experimental use only. Not for resale without express authorization.)
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