Rab5A Antibody | Affinity Biosciences

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Figure 8 (a) RAB5A overexpression and shRNA plasmid transfection HCAECs in vitro.

Figure 10 (a, d) Wound healing assay in all groups.

Product:	Rab5A Antibody
Catalog:	DF6314
Description:	Rabbit polyclonal antibody to Rab5A
Application:	WB IHC
Reactivity:	Human, Mouse, Rat
Prediction:	Bovine, Horse, Rabbit, Dog, Chicken
Mol.Wt.:	24kDa; 24kD(Calculated).
Uniprot:	P20339
RRID:	AB_2838280

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Blocking peptides are peptides that bind specifically to the target antibody and block antibody binding. These peptide usually contains the epitope recognized by the antibody. Antibodies bound to the blocking peptide no longer bind to the epitope on the target protein. By comparing the staining from the blocked antibody versus the antibody alone, one can see which staining is specific.

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200ul	$350	In stock

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Related Downloads

MSDS

Data Sheet

COA

Protocols

WB Handbook

IHC Protocol

IF/ICC Protocol

Product Info

Source:

Rabbit

Application:

WB 1:500-1:2000, IHC 1:50-1:200
*The optimal dilutions should be determined by the end user.
*Tips:

WB: For western blot detection of denatured protein samples. IHC: For immunohistochemical detection of paraffin sections (IHC-p) or frozen sections (IHC-f) of tissue samples. IF/ICC: For immunofluorescence detection of cell samples. ELISA(peptide): For ELISA detection of antigenic peptide.

Reactivity:

Human,Mouse,Rat

Prediction:

Bovine(88%), Horse(100%), Rabbit(100%), Dog(100%), Chicken(88%)

Clonality:

Polyclonal

Specificity:

Rab5A Antibody detects endogenous levels of total Rab5A.

RRID:

AB_2838280
Cite Format: Affinity Biosciences Cat# DF6314, RRID:AB_2838280.

Conjugate:

Unconjugated.

Purification:

The antiserum was purified by peptide affinity chromatography using SulfoLink™ Coupling Resin (Thermo Fisher Scientific).

Storage:

Rabbit IgG in phosphate buffered saline , pH 7.4, 150mM NaCl, 0.02% sodium azide and 50% glycerol. Store at -20 °C. Stable for 12 months from date of receipt.

Alias:

Fold/Unfold

RAB 5; RAB 5A; RAB5A; RAB5A member RAS oncogene family; RAB5A_HUMAN; RAS associated protein RAB5A; Ras related protein Rab 5A; Ras-related protein Rab-5A;

Immunogens

Immunogen:

Uniprot:

P20339(RAB5A_HUMAN) >>Visit HPA database.

Gene(ID):

RAB5A(5868)

Description:

Rab5 is a member of the Ras superfamily of small Rab GTPases. Rab5 is localized at the plasma membrane and early endosomes and functions as a key regulator of vesicular trafficking during early endocytosis (1). The conformational change between Rab5 GTP/GDP states is essential for its biological function as a rate limiting regulator at multiple steps during endocytosis (1,2). Rab5 exerts its function by interacting with several Rab5-specific effectors (1-3). These proteins form complexes with Rab5 on a specialized Rab domain of the endosome and promote recycling of Rab5-cargo targets between endosome and the plasma membrane.

Sequence:

P20339 RAB5A_HUMAN:
Protein BLAST With
NCBI/
ExPASy/
Uniprot

MASRGATRPNGPNTGNKICQFKLVLLGESAVGKSSLVLRFVKGQFHEFQESTIGAAFLTQTVCLDDTTVKFEIWDTAGQERYHSLAPMYYRGAQAAIVVYDITNEESFARAKNWVKELQRQASPNIVIALSGNKADLANKRAVDFQEAQSYADDNSLLFMETSAKTSMNVNEIFMAIAKKLPKNEPQNPGANSARGRGVDLTEPTQPTRNQCCSN

Predictions

Predictions:

Score>80(red) has high confidence and is suggested to be used for WB detection. *The prediction model is mainly based on the alignment of immunogen sequences, the results are for reference only, not as the basis of quality assurance.

Species

Results

Score

Horse

100

Dog

100

Rabbit

100

Bovine

Chicken

Pig

Sheep

Xenopus

Zebrafish

Model Confidence:
High(score>80) Medium(80>score>50) Low(score<50) No confidence

PTMs - P20339 As Substrate

P20339

Site	PTM Type	Enzyme	Source
S3	Phosphorylation		Uniprot
R4	Methylation		Uniprot
T7	Phosphorylation	Q02156 (PRKCE)	Uniprot
K22	Acetylation		Uniprot
S29	Phosphorylation		Uniprot
K33	Ubiquitination		Uniprot
Y82	Phosphorylation		Uniprot
S84	Phosphorylation		Uniprot
K116	Ubiquitination		Uniprot
S123	Phosphorylation	P27361 (MAPK3)	Uniprot
K134	Ubiquitination		Uniprot
K140	Ubiquitination		Uniprot
R197	Methylation		Uniprot
T202	Phosphorylation		Uniprot
T205	Phosphorylation		Uniprot
T208	Phosphorylation		Uniprot

Research Backgrounds

P20339_RAB5A_HUMAN

Function:

Small GTPase which cycles between active GTP-bound and inactive GDP-bound states. In its active state, binds to a variety of effector proteins to regulate cellular responses such as of intracellular membrane trafficking, from the formation of transport vesicles to their fusion with membranes. Active GTP-bound form is able to recruit to membranes different sets of downstream effectors directly responsible for vesicle formation, movement, tethering and fusion. RAB5A is required for the fusion of plasma membranes and early endosomes. Contributes to the regulation of filopodia extension. Required for the exosomal release of SDCBP, CD63, PDCD6IP and syndecan. Regulates maturation of apoptotic cell-containing phagosomes, probably downstream of DYN2 and PIK3C3 (By similarity).

PTMs:

Phosphorylation of Ser-84 in the switch II region by LRRK2 prevents the association of RAB regulatory proteins, including RAB GDP dissociation inhibitors GDI1 and GDI2.

Subcellular Location:

Cell membrane>Lipid-anchor>Cytoplasmic side. Early endosome membrane>Lipid-anchor. Melanosome. Cytoplasmic vesicle. Cell projection>Ruffle. Membrane. Cytoplasm>Cytosol. Cytoplasmic vesicle>Phagosome membrane. Endosome membrane.
Note: Enriched in stage I melanosomes (PubMed:17081065). Alternates between membrane-bound and cytosolic forms (Probable).

Subunit Structure:

Interacts with SGSM1 and SGSM3 (By similarity). Interacts with PIK3CB (By similarity). Interacts with GDI1; this promotes dissociation from membranes; phosphorylation at Ser-84 disrupts this interaction. Interacts with GDI2; phosphorylation at Ser-84 disrupts the interaction. Interacts with EEA1. Interacts with RIN1 and GAPVD1, which regulate its pathway, probably by acting as a GEF. Interacts with RINL. Interacts with ALS2CL, SUN2, ZFYVE20 and RUFY1. Interacts with RABEP1; one RABEP1 homodimer binds two RAB5A chains, but at opposite sides of the dimer. Interacts with OCRL. Interacts with INPP5F. May be a component of a complex composed of RAB5A, DYN2 and PIK3C3 (By similarity). Does not interact with BLOC-3 complex (heterodimer of HPS1 and HPS4) (By similarity). Interacts with CLN5. Interacts with APPL2 (By similarity). Interacts with F8A1/F8A2/F8A3 (By similarity). Found in a complex with F8A1/F8A2/F8A3, HTT and RAB5A; mediates the recruitment of HTT by RAB5A onto early endosomes (By similarity).

Family&Domains:

Belongs to the small GTPase superfamily. Rab family.

Research Fields

· Cellular Processes > Transport and catabolism > Endocytosis. (View pathway)

· Cellular Processes > Transport and catabolism > Phagosome. (View pathway)

· Environmental Information Processing > Signal transduction > Ras signaling pathway. (View pathway)

· Human Diseases > Neurodegenerative diseases > Amyotrophic lateral sclerosis (ALS).

· Human Diseases > Infectious diseases: Parasitic > Amoebiasis.

· Human Diseases > Infectious diseases: Bacterial > Tuberculosis.

· Organismal Systems > Excretory system > Vasopressin-regulated water reabsorption.

References

1). Identification of Key Genes Associated with Endothelial Cell Dysfunction in Atherosclerosis Using Multiple Bioinformatics Tools. Biomed Research International, 2022 (PubMed: 35059464)

Application: WB Species: Human Sample: HCAECs

Figure 8 (a) RAB5A overexpression and shRNA plasmid transfection HCAECs in vitro. There was no significant difference in transfection efficiency between two groups (28.7% and 27.3%), and the green fluorescence carried by the plasmid itself can be seen. (b, c) The results of measuring the expression of RAB5A protein after transfection are shown in (b). Relative quantitative comparison of western blot analyses in (C). ∗p < 0.05, compared to the HCAEC group; #p < 0.05, compared to the HCAEC-RAB5A overexpression group.

Application: IF/ICC Species: Human Sample: HCAECs

Figure 10 (a, d) Wound healing assay in all groups. (a) Migration distance of HCAECs in different conditions. (d) Relative quantitative comparison of healing speed analyses mentioned in (a). (b, e) Transwell migration assay in all groups. (b) Representative photomicrographs showing migrating cells in different conditions. (e) Quantification of number of migrating cells demonstrated that overexpression of RAB5A promoted HCAEC migration function (n = 6/group). (c, f) Tube formation assay. (c) Representative photomicrographs showing tube formation of HCAECs in different conditions. (f) Quantification of HCAEC tube junction numbers demonstrated that overexpression of RAB5A ameliorated HCAECs to form capillary-like structures (n = 6/group). ∗p < 0.05, compared to the HCAEC group; #p < 0.05, compared to the HCAEC-RAB5A overexpression group.

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Rab5A Antibody - #DF6314