PREX1 Antibody [J20N16] | Primary Antibodies

Application: Reactivity: Human

Usage Information

Dilution
IHC1:150

Application
IHC

Reactivity
Human

Source
Rabbit Monoclonal Antibody

Storage Buffer
PBS, pH 7.2+50% Glycerol+0.05% BSA+0.01% NaN3

Storage (from the date of receipt)
-20°C (avoid freeze-thaw cycles), 2 years

Predicted MW Observed MW
186 kDa 190 kDa, 110 kDa
^*Why do the predicted and actual molecular weights differ? The following reasons may explain differences between the predicted and actual protein molecular weight. Post-translational modifications(e.g., phosphorylation, glycosylation); Splice variants and isoforms; Relative charge; Multimerization.

Positive Control	Human colon carcinoma; Human lung carcinoma; Human lymphoma; Human melanoma; T-47D cells
Negative Control	MCF 10A cells

Experimental Methods

IHC
Experimental Protocol： Deparaffinization/Rehydration 1. Deparaffinize/hydrate sections: 2. Incubate sections in three washes of xylene for 5 min each. 3. Incubate sections in two washes of 100% ethanol for 10 min each. 4. Incubate sections in two washes of 95% ethanol for 10 min each. 5. Wash sections two times in dH2O for 5 min each. 6.Antigen retrieval: For Citrate: Heat slides in a microwave submersed in 1X citrate unmasking solution until boiling is initiated; continue with 10 min at a sub-boiling temperature (95°-98°C). Cool slides on bench top for 30 min. Staining 1. Wash sections in dH2O three times for 5 min each. 2. Incubate sections in 3% hydrogen peroxide for 10 min. 3. Wash sections in dH2O two times for 5 min each. 4. Wash sections in wash buffer for 5 min. 5. Block each section with 100–400 µl of blocking solution for 1 hr at room temperature. 6. Remove blocking solution and add 100–400 µl primary antibody diluent in to each section. Incubate overnight at 4°C. 7. Remove antibody solution and wash sections with wash buffer three times for 5 min each. 8. Cover section with 1–3 drops HRPas needed. Incubate in a humidified chamber for 30 min at room temperature. 9. Wash sections three times with wash buffer for 5 min each. 10. Add DAB Chromogen Concentrate to DAB Diluent and mix well before use. 11. Apply 100–400 µl DAB to each section and monitor closely. 1–10 min generally provides an acceptable staining intensity. 12. Immerse slides in dH2O. 13. If desired, counterstain sections with hematoxylin. 14. Wash sections in dH2O two times for 5 min each. 15. Dehydrate sections: Incubate sections in 95% ethanol two times for 10 sec each; Repeat in 100% ethanol, incubating sections two times for 10 sec each; Repeat in xylene, incubating sections two times for 10 sec each. 16. Mount sections with coverslips and mounting medium.

IHC

Experimental Protocol：

Deparaffinization/Rehydration

1. Deparaffinize/hydrate sections:

2. Incubate sections in three washes of xylene for 5 min each.

3. Incubate sections in two washes of 100% ethanol for 10 min each.

4. Incubate sections in two washes of 95% ethanol for 10 min each.

5. Wash sections two times in dH2O for 5 min each.

6.Antigen retrieval: For Citrate: Heat slides in a microwave submersed in 1X citrate unmasking solution until boiling is initiated; continue with 10 min at a sub-boiling temperature (95°-98°C). Cool slides on bench top for 30 min.

Staining

1. Wash sections in dH2O three times for 5 min each.

2. Incubate sections in 3% hydrogen peroxide for 10 min.

3. Wash sections in dH2O two times for 5 min each.

4. Wash sections in wash buffer for 5 min.

5. Block each section with 100–400 µl of blocking solution for 1 hr at room temperature.

6. Remove blocking solution and add 100–400 µl primary antibody diluent in to each section. Incubate overnight at 4°C.

7. Remove antibody solution and wash sections with wash buffer three times for 5 min each.

8. Cover section with 1–3 drops HRPas needed. Incubate in a humidified chamber for 30 min at room temperature.

9. Wash sections three times with wash buffer for 5 min each.

10. Add DAB Chromogen Concentrate to DAB Diluent and mix well before use.

11. Apply 100–400 µl DAB to each section and monitor closely. 1–10 min generally provides an acceptable staining intensity.

12. Immerse slides in dH2O.

13. If desired, counterstain sections with hematoxylin.

14. Wash sections in dH2O two times for 5 min each.

15. Dehydrate sections: Incubate sections in 95% ethanol two times for 10 sec each; Repeat in 100% ethanol, incubating sections two times for 10 sec each; Repeat in xylene, incubating sections two times for 10 sec each.

16. Mount sections with coverslips and mounting medium.

Datasheet & SDS

Biological Description

Specificity
PREX1 Antibody (Rabbit mAb) [J20N16] detects endogenous levels of total PREX1 protein.

Uniprot ID
Q8TCU6

Clone
J20N16

Synonym(s)
KIAA1415, P-Rex1, phosphatidylinositol-3,4,5-trisphosphate-dependent Rac exchange factor 1, PIP3 dependent Rac exchange factor 1, PREX1, PtdIns(3,4,5)-dependent Rac exchanger 1

Background
PREX1, also known as phosphatidylinositol‑3,4,5‑trisphosphate‑dependent Rac exchanger 1, belongs to the Dbl family of Rho‑type guanine nucleotide exchange factors and functions as a Rac‑directed signal integrator downstream of phosphoinositide 3‑kinase and G protein‑coupled receptors. The protein contains an N‑terminal Dbl‑homology catalytic domain paired with a pleckstrin homology domain that senses phosphatidylinositol‑3,4,5‑trisphosphate, flanked by tandem DEP domains and C‑terminal PDZ and IP4P‑like regions that assemble regulatory and effector interactions and support long‑range allosteric control across a two‑layered autoinhibited architecture. Binding of phosphatidylinositol‑3,4,5‑trisphosphate to the pleckstrin homology domain and engagement of Gβγ subunits at defined regulatory surfaces release intramolecular restraints within PREX1, expose the Dbl‑homology active site, and permit efficient GDP–GTP exchange on Rac‑family GTPases, which elevates Rac‑GTP levels and couples receptor inputs to the core Rho GTPase network. Activated Rac then links PREX1 output to downstream effectors such as p21‑activated kinases and other Rac‑responsive kinases that remodel the actin cytoskeleton, regulate lamellipodia dynamics, and adjust adhesion complexes, placing PREX1 as a central controller of directed migration, invasion, and cell–matrix interactions in contexts where G protein‑coupled receptor and receptor tyrosine kinase signals converge. Structural analysis of full‑length PREX1 shows that the catalytic module packs against distal domains in an autoinhibitory configuration and that coincident lipid and Gβγ binding induces large conformational rearrangements across both layers of the molecule, aligning the Dbl‑homology domain with Rac substrates and stabilizing an active signaling conformation at the plasma membrane. Rac activation driven by PREX1 contributes to pathways that connect to ERK signaling and other mitogenic cascades, linking this exchange factor to regulation of proliferation and survival in addition to motility, and placing it within broader signaling circuits that integrate chemokine receptors, sphingosine‑1‑phosphate receptors, and growth factor receptors. Elevated PREX1 expression and activation are associated with increased invasive behavior and metastatic potential in cancers such as breast cancer, prostate cancer, melanoma, and glioblastoma, where PREX1–Rac signaling shapes cytoskeletal organization, directional migration, and matrix invasion, and where Rac‑dependent kinase outputs feed into transcriptional programs that support tumor progression.

Background

PREX1, also known as phosphatidylinositol‑3,4,5‑trisphosphate‑dependent Rac exchanger 1, belongs to the Dbl family of Rho‑type guanine nucleotide exchange factors and functions as a Rac‑directed signal integrator downstream of phosphoinositide 3‑kinase and G protein‑coupled receptors. The protein contains an N‑terminal Dbl‑homology catalytic domain paired with a pleckstrin homology domain that senses phosphatidylinositol‑3,4,5‑trisphosphate, flanked by tandem DEP domains and C‑terminal PDZ and IP4P‑like regions that assemble regulatory and effector interactions and support long‑range allosteric control across a two‑layered autoinhibited architecture. Binding of phosphatidylinositol‑3,4,5‑trisphosphate to the pleckstrin homology domain and engagement of Gβγ subunits at defined regulatory surfaces release intramolecular restraints within PREX1, expose the Dbl‑homology active site, and permit efficient GDP–GTP exchange on Rac‑family GTPases, which elevates Rac‑GTP levels and couples receptor inputs to the core Rho GTPase network. Activated Rac then links PREX1 output to downstream effectors such as p21‑activated kinases and other Rac‑responsive kinases that remodel the actin cytoskeleton, regulate lamellipodia dynamics, and adjust adhesion complexes, placing PREX1 as a central controller of directed migration, invasion, and cell–matrix interactions in contexts where G protein‑coupled receptor and receptor tyrosine kinase signals converge. Structural analysis of full‑length PREX1 shows that the catalytic module packs against distal domains in an autoinhibitory configuration and that coincident lipid and Gβγ binding induces large conformational rearrangements across both layers of the molecule, aligning the Dbl‑homology domain with Rac substrates and stabilizing an active signaling conformation at the plasma membrane. Rac activation driven by PREX1 contributes to pathways that connect to ERK signaling and other mitogenic cascades, linking this exchange factor to regulation of proliferation and survival in addition to motility, and placing it within broader signaling circuits that integrate chemokine receptors, sphingosine‑1‑phosphate receptors, and growth factor receptors. Elevated PREX1 expression and activation are associated with increased invasive behavior and metastatic potential in cancers such as breast cancer, prostate cancer, melanoma, and glioblastoma, where PREX1–Rac signaling shapes cytoskeletal organization, directional migration, and matrix invasion, and where Rac‑dependent kinase outputs feed into transcriptional programs that support tumor progression.

References
https://pubmed.ncbi.nlm.nih.gov/25284585/ https://pubmed.ncbi.nlm.nih.gov/27358402/

Protein Size (kDa)	PVDF Transfer Membrane Pore Size (μm)
< 10	0.22
10-20	0.22
20-30	0.45
30-45	0.45
45-70	0.45
80-100	0.45
100-140	0.45
> 140	0.45

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Protein Size (kDa)	Separating Gel Percentage
4-40	20%
12-45	15%
10-70	12.5%
15-100	10%
25-100	8%
60-210	5%

PREX1 Antibody (Rabbit mAb) [J20N16]

Usage Information

Experimental Methods

Datasheet & SDS

Biological Description

Tech Support