MA4-001

antibody from Invitrogen Antibodies

Targeting: RASA1 CM-AVM, GAP, p120, p120GAP, p120RASGAP, RASA

Western blot (Supportive data in Antibodypedia)

Immunocytochemistry (Supportive data in Antibodypedia)

Immunoprecipitation (Recommended by provider)

Antibody Data

Product number

MA4-001

Provider

Invitrogen Antibodies

Product name

RASA1 Monoclonal Antibody (B4F8)

Antibody type

Monoclonal

Antigen

Recombinant full-length protein

Description

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.

Reactivity

Human, Mouse, Rat, Bovine

Host

Mouse

Isotype

IgG

Antibody clone number

B4F8

Vial size

100 µg

Concentration

1 mg/mL

Storage

-20° C, Avoid Freeze/Thaw Cycles

References

Submitted references

Differential expression of cytoskeletal regulatory factors in the adolescent prefrontal cortex: Implications for cortical development.

Shapiro LP, Parsons RG, Koleske AJ, Gourley SL
Journal of neuroscience research 2017 May;95(5):1123-1143

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Molecular cloning of cDNAs encoding the GAP-associated protein p190: implications for a signaling pathway from ras to the nucleus.

Settleman J, Narasimhan V, Foster LC, Weinberg RA
Cell 1992 May 1;69(3):539-49

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Molecular cloning of cDNAs encoding the GAP-associated protein p190: implications for a signaling pathway from ras to the nucleus.

Settleman J, Narasimhan V, Foster LC, Weinberg RA
Cell 1992 May 1;69(3):539-49

Western blot

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Western blot analysis of GTPase Activating Protein was performed by loading 25 µg 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.

Immunocytochemistry

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

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 fluorescent red phalloidin and nuclei (blue) were stained with DAPI for 5-10 minutes in the dark. Images were taken at a magnification of 60x.

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

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 fluorescent red phalloidin and nuclei (blue) were stained with DAPI for 5-10 minutes in the dark. Images were taken at a magnification of 60x.

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

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 fluorescent red phalloidin and nuclei (blue) were stained with DAPI for 5-10 minutes in the dark. Images were taken at a magnification of 60x.