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GEF H1 Recombinant Antibody

Paraformaldehyde-fixed, paraffin embedded (rat testis); Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15min; Block endogenous peroxidase by 3% hydrogen peroxide for 20 minutes; Blocking buffer (normal goat serum) at 37\u00b0C for 30min; Incubation with (ARHGEF2 (9D4)) Monoclonal Antibody, Unconjugated (bsm-54369R) at 1:50 overnight at 4\u00b0C, followed by operating according to SP Kit(Rabbit) (sp-0023) instructionsand DAB staining.
Paraformaldehyde-fixed, paraffin embedded (rat testis); Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15min; Block endogenous peroxidase by 3% hydrogen peroxide for 20 minutes; Blocking buffer (normal goat serum) at 37\u00b0C for 30min; Incubation with (ARHGEF2 (9D4)) Monoclonal Antibody, Unconjugated (bsm-54369R) at 1:50 overnight at 4\u00b0C, followed by operating according to SP Kit(Rabbit) (sp-0023) instructionsand DAB staining.
Paraformaldehyde-fixed, paraffin embedded (human gastric carcinoma); Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15min; Block endogenous peroxidase by 3% hydrogen peroxide for 20 minutes; Blocking buffer (normal goat serum) at 37\u00b0C for 30min; Incubation with (ARHGEF2 (9D4)) Monoclonal Antibody, Unconjugated (bsm-54369R) at 1:50 overnight at 4\u00b0C, followed by operating according to SP Kit(Rabbit) (sp-0023) instructionsand DAB staining.
Paraformaldehyde-fixed, paraffin embedded (human gastric carcinoma); Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15min; Block endogenous peroxidase by 3% hydrogen peroxide for 20 minutes; Blocking buffer (normal goat serum) at 37\u00b0C for 30min; Incubation with (ARHGEF2 (9D4)) Monoclonal Antibody, Unconjugated (bsm-54369R) at 1:50 overnight at 4\u00b0C, followed by operating according to SP Kit(Rabbit) (sp-0023) instructionsand DAB staining.
Lane 1: Human HeLa cell lysates; Lane 2: Human MCF-7 cell lysates probed with GEF H Monoclonal Antibody, Unconjugated (bsm-54369R) at 1:500 dilution and 4\u00b0C overnight incubation. Followed by conjugated secondary antibody incubation at room temperature for 60 min.
Lane 1: Human HeLa cell lysates; Lane 2: Human MCF-7 cell lysates probed with GEF H Monoclonal Antibody, Unconjugated (bsm-54369R) at 1:500 dilution and 4\u00b0C overnight incubation. Followed by conjugated secondary antibody incubation at room temperature for 60 min.
The HeLa (H) cells were fixed with 4% PFA (10 min at r.t.) and then permeabilized with 90% ice-cold methanol for 20 min at -20\u2103,the cells then were incubated in 5%BSA to block non-specific protein-protein interactions (30 min at r.t.), followed by secondary antibody incubation for 40 min at room temperature. Primary Antibody (green):Rabbit Anti-GEF H1 antibody (bsm-54369R,1:100); Isotype Control (orange): Rabbit IgG (bs-0295P). Blank control (black): PBS. Acquisition of 20,000 events was performed.
The HeLa (H) cells were fixed with 4% PFA (10 min at r.t.) and then permeabilized with 90% ice-cold methanol for 20 min at -20\u2103,the cells then were incubated in 5%BSA to block non-specific protein-protein interactions (30 min at r.t.), followed by secondary antibody incubation for 40 min at room temperature. Primary Antibody (green):Rabbit Anti-GEF H1 antibody (bsm-54369R,1:100); Isotype Control (orange): Rabbit IgG (bs-0295P). Blank control (black): PBS. Acquisition of 20,000 events was performed.
4% Paraformaldehyde-fixed HeLa (H) cell; Triton X-100 at r.t. for 20 min; Antibody incubation with (GEF H1) monoclonal Antibody, unconjugated (bsm-543691R) 1:100, 90 min at 37\u00b0C; followed by BF488 conjugated Goat Anti-Rabbit IgG antibody (green, bs-60295G-BF488) at 37\u00b0C for 90 min, DAPI (blue, C02-04002) was used to stain the cell nuclei. PBS instead of the primary antibody was used as the blank control.
4% Paraformaldehyde-fixed HeLa (H) cell; Triton X-100 at r.t. for 20 min; Antibody incubation with (GEF H1) monoclonal Antibody, unconjugated (bsm-543691R) 1:100, 90 min at 37\u00b0C; followed by BF488 conjugated Goat Anti-Rabbit IgG antibody (green, bs-60295G-BF488) at 37\u00b0C for 90 min, DAPI (blue, C02-04002) was used to stain the cell nuclei. PBS instead of the primary antibody was used as the blank control.
Bioss unconjugated antibody vial

Applications

  • WB
  • FCM
  • IHC-P
  • IF(IHC-P)
  • IF(ICC)

Reactivity

  • Human

Predicted Reactivity

  • Mouse
  • Rat
Overview
Catalog # bsm-54369r-100uL
Product Name GEF H1 Recombinant Antibody
Applications
WB FCM IHC-P IF(IHC-P) IF(ICC)
Reactivity Human
Predicted Reactivity Mouse, Rat
Specifications
Conjugation Unconjugated
Host Rabbit
Source Recombinant protein corresponding to C-terminal human GEF H1.
Clonality Recombinant
Isotype IgG
Concentration Lot Dependent
Purification Purified by Protein A.
Storage Buffer 0.01M TBS (pH 7.4), 1% BSA, 0.02% Proclin 300, and 50% Glycerol
Storage Condition Shipped at 4C. Store at -20C for one year. Avoid repeated freeze/thaw cycles.
Target
Gene ID 9181
Swiss Prot Q92974
Subcellular location Cell membrane, Golgi apparatus, Cytoskeleton
Synonyms Rho guanine nucleotide exchange factor 2; Guanine nucleotide exchange factor H1; Microtubule-regulated Rho-GEF; Proliferating cell nucleolar antigen p40; GEF-H1; ARHGEF2; KIAA0651; LFP40
Background Activates Rho-GTPases by promoting the exchange of GDP for GTP. May be involved in epithelial barrier permeability, cell motility and polarization, dendritic spine morphology, antigen presentation, leukemic cell differentiation, cell cycle regulation, innate immune response, and cancer. Binds Rac-GTPases, but does not seem to promote nucleotide exchange activity toward Rac-GTPases. May stimulate instead the cortical activity of Rac. Inactive toward CDC42, TC10, or Ras-GTPases. Forms an intracellular sensing system along with NOD1 for the detection of microbial effectors during cell invasion by pathogens. Required for RHOA and RIP2 dependent NF-kappaB signaling pathways activation upon S.flexneri cell invasion. Involved not only in sensing peptidoglycan (PGN)-derived muropeptides through NOD1 that is independent of its GEF activity, but also in the activation of NF-kappaB by Shigella effector proteins (IpgB2 and OspB) which requires its GEF activity and the activation of RhoA. Involved in innate immune signaling transduction pathway promoting cytokine IL6/interleukin-6 and TNF-alpha secretion in macrophage upon stimulation by bacterial peptidoglycans; acts as a signaling intermediate between NOD2 receptor and RIPK2 kinase. Contributes to the tyrosine phosphorylation of RIPK2 through Src tyrosine kinase leading to NF-kappaB activation by NOD2.
Application Dilution
WB 1:300-5000
FCM 1:20-100
IHC-P 1:200-400
IF(IHC-P) 1:50-200
IF(ICC) 1:50-200

Frequently asked questions

Which applications is GEF H1 Recombinant Antibody (bsm-54369R) validated for, and at what dilution?

bsm-54369R is validated for WB (1:300-5000); FCM (1:20-100); IHC-P (1:200-400); IF(IHC-P) (1:50-200); IF(ICC) (1:50-200). These are recommended starting dilutions; optimal dilutions should be determined by the end user.

Which species does GEF H1 Recombinant Antibody (bsm-54369R) react with?

Tested reactivity: Human. Predicted reactivity (expected from sequence homology but not yet tested): Mouse, Rat.

How should bsm-54369R be stored?

Shipped at 4C. Store at -20C for one year. Avoid repeated freeze/thaw cycles. Storage buffer: 0.01M TBS (pH 7.4), 1% BSA, 0.02% Proclin 300, and 50% Glycerol

What immunogen was used to raise GEF H1 Recombinant Antibody (bsm-54369R)?

Recombinant protein corresponding to C-terminal human GEF H1.

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US only. One sample per paid order, subject to review. Backorders may ship separately. Confirm Free at checkout. Check application and species suitability; same target does not guarantee equivalent performance. For research use only.

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