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Human RAB8 ELISA Kit is a single-wash 90-min SimpleStep ELISA® for the quantitative measurement of Human RAB8 in Cell Lysate samples.

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Images

Sandwich ELISA - Human RAB8 ELISA Kit (AB255718), expandable thumbnail
  • Sandwich ELISA - Human RAB8 ELISA Kit (AB255718), expandable thumbnail

Key facts

Detection method

Colorimetric

Sample types

Cell Lysate

Assay type

Sandwich (quantitative)

Reactive species

Human

Range

0.09 - 6 ng/mL

Assay time

1h 30m

Sensitivity

= 0.024 ng/mL

Reactivity data

Application

sELISA

Reactivity

Reacts

Dilution info
-
Notes

-

Associated Products

Select an associated product type

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1 product for Alternative Version

Target data

Function

The small GTPases Rab are key regulators of intracellular membrane trafficking, from the formation of transport vesicles to their fusion with membranes. Rabs cycle between an inactive GDP-bound form and an active GTP-bound form that is able to recruit to membranes different sets of downstream effectors directly responsible for vesicle formation, movement, tethering and fusion. That Rab is involved in polarized vesicular trafficking and neurotransmitter release. Together with RAB11A, RAB3IP, the exocyst complex, PARD3, PRKCI, ANXA2, CDC42 and DNMBP promotes transcytosis of PODXL to the apical membrane initiation sites (AMIS), apical surface formation and lumenogenesis (PubMed:20890297). Regulates the compacted morphology of the Golgi (PubMed:26209634). Together with MYO5B and RAB11A participates in epithelial cell polarization (PubMed:21282656). Also involved in membrane trafficking to the cilium and ciliogenesis (PubMed:21844891, PubMed:30398148). Together with MICALL2, may also regulate adherens junction assembly (By similarity). May play a role in insulin-induced transport to the plasma membrane of the glucose transporter GLUT4 and therefore play a role in glucose homeostasis (By similarity). Involved in autophagy (PubMed:27103069). Participates in the export of a subset of neosynthesized proteins through a Rab8-Rab10-Rab11-dependent endososomal export route (PubMed:32344433). Targeted to and stabilized on stressed lysosomes through LRRK2 phosphorylation (PubMed:30209220). Suppresses stress-induced lysosomal enlargement through EHBP1 and EHNP1L1 effector proteins (PubMed:30209220).

Alternative names

What's included?

1 x 96 Tests
Components
10X Human RAB8 Capture Antibody
1 x 600 µL
10X Human RAB8 Detector Antibody
1 x 600 µL
10X Wash Buffer PT (ab206977)
1 x 20 mL
50X Cell Extraction Enhancer Solution (ab193971)
1 x 1 mL
5X Cell Extraction Buffer PTR (ab193970)
1 x 10 mL
Antibody Diluent 4BR
1 x 6 mL
Human RAB8 Lyophilized Recombinant Protein
2 x 1 Vial
Plate Seals
1 x 1 Unit
Sample Diluent NS (ab193972)
1 x 12 mL
SimpleStep Pre-Coated 96-Well Microplate (ab206978)
1 x 1 Unit
Stop Solution
1 x 12 mL
TMB Development Solution
1 x 12 mL

Recommended products

Human RAB8 ELISA Kit is a single-wash 90-min SimpleStep ELISA® for the quantitative measurement of Human RAB8 in Cell Lysate samples.

Key facts

Detection method

Colorimetric

Sample types

Cell Lysate

Assay type

Sandwich (quantitative)

Reactive species

Human

Range

0.09 - 6 ng/mL

Assay time

1h 30m

Assay Platform

Pre-coated microplate (12 x 8 well strips)

Sensitivity

= 0.024 ng/mL

Precision

Intra assay

Sample

Extract

n
8
mean
-
SD
-
C.V.

3

Inter assay

Sample

Extract

n
3
mean
-
SD
-
C.V.

2.2

Recovery

Sample specific recovery

Sample type

Cell Lysate

Average %

= 87

Range

81 - 93 %

Sample type

Cell culture extracts

Average %

= 95

Range

93 - 98 %

Sample type

Tissue Extracts

Average %

= 102

Range

100 - 105 %

Storage

Shipped at conditions

Blue Ice

Appropriate short-term storage conditions

+4°C

Appropriate long-term storage conditions

+4°C

Storage information

+4°C

Notes

Human RAB8 ELISA Kit (ab255718) is a single-wash 90 min sandwich ELISA designed for the quantitative measurement of RAB8 protein in cell lysate. It uses our proprietary SimpleStep ELISA® technology. Quantitate Human RAB8 with 0.024 ng/ml sensitivity.

SimpleStep ELISA® technology employs capture antibodies conjugated to an affinity tag that is recognized by the monoclonal antibody used to coat our SimpleStep ELISA® plates. This approach to sandwich ELISA allows the formation of the antibody-analyte sandwich complex in a single step, significantly reducing assay time. See the SimpleStep ELISA® protocol summary in the image section for further details. Our SimpleStep ELISA® technology provides several benefits:

- Single-wash protocol reduces assay time to 90 minutes or less
- High sensitivity, specificity and reproducibility from superior antibodies
- Fully validated in biological samples
- 96-wells plate breakable into 12 x 8 wells strips

A 384-well SimpleStep ELISA® microplate (Pre-coated 384 well Microplate SimpleStep ELISA® ab203359) is available to use as an alternative to the 96-well microplate provided with SimpleStep ELISA® kits.

The small GTPases Rab are key regulators of intracellular membrane trafficking, from the formation of transport vesicles to their fusion with membranes. Rabs cycle between an inactive GDP-bound form and an active GTP-bound form that is able to recruit to membranes different sets of downstream effectors directly responsible for vesicle formation, movement, tethering, and fusion. Rab proteins are highly conserved across species; human RAB8, also known as Ras-related protein Rab-8A, is 97.5% identical on amino acids level to mouse RAB8. Mouse and rat RAB8 proteins are 100% identical.

Abcam has not and does not intend to apply for the REACH Authorisation of customers' uses of products that contain European Authorisation list (Annex XIV) substances.
It is the responsibility of our customers to check the necessity of application of REACH Authorisation, and any other relevant authorisations, for their intended uses.

Supplementary info

This supplementary information is collated from multiple sources and compiled automatically.

Activity summary

RAB8A is a small GTPase also referred to as Ras-related protein Rab-8A involved in intracellular membrane trafficking. It plays a critical role in the regulation of exocytosis the process by which cells release substances to the extracellular space. RAB8A mainly functions by cycling between an inactive GDP-bound form and an active GTP-bound form facilitating vesicle budding transport and fusion with target membranes. This protein is expressed in various tissues including the brain and pancreas where it assists in important cellular functions. RAB8A has a molecular mass of approximately 24 kDa.

Biological function summary

The small GTPase RAB8A contributes to the biosynthetic transport from the trans-Golgi network to the plasma membrane. It performs this activity in coordination with other proteins as part of a larger complex. RAB8A also plays an essential role in cilia formation and maintenance affecting cell polarity and signaling functions. The protein's ability to regulate vesicle movement makes it important for cellular homeostasis and communication.

Pathways

The regulatory function of RAB8A is significant in the ciliary membrane trafficking pathway and the insulin signaling pathway. Within these pathways RAB8A interacts closely with other proteins like Rab11 and Rabin8 which help support its roles in directing vesicle transport. These interactions are essential for proper cellular signaling and the maintenance of polarized cellular structures further supporting the function of various cellular pathways.

Associated diseases and disorders

The dysfunction of RAB8A has associations with certain ciliopathies notably Bardet-Biedl syndrome. This condition involves abnormalities in cilia structure and function leading to symptoms like kidney disease vision impairment and obesity. Moreover disruptions in RAB8A have connections with insulin resistance implicating it in metabolic disorders. In the context of these diseases proteins such as BBS proteins and Rab11 play a role in mediating the effects of dysfunctional RAB8A-related pathways.

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2 product images

  • Sandwich ELISA - Human RAB8 ELISA Kit (ab255718), expandable thumbnail

    Sandwich ELISA - Human RAB8 ELISA Kit (ab255718)

    Example of human RAB8 standard curve in 1X Cell Extraction Buffer PTR.

    The RAB8 standard curve was prepared as described in Section 10. Raw data values are shown in the table. Background-subtracted data values (mean +/- SD) are graphed.

  • Sandwich ELISA - Human RAB8 ELISA Kit (ab255718), expandable thumbnail

    Sandwich ELISA - Human RAB8 ELISA Kit (ab255718)

    Interpolated concentrations of native RAB8 in human HeLa extract based on a 25 μg/mL extract, HepG2 cell extract based on a 50 μg/mL extract, and human colon extract based on a 250 μg/mL extract.

    The concentrations of RAB8 were measured in duplicate and interpolated from the RAB8 standard curve and corrected for sample dilution. The interpolated dilution factor corrected values are plotted (mean +/- SD, n=2). The mean RAB8 concentration was determined to be 2.16 ng/mL in HeLa cell extract, 1.16 ng/mL HepG2 cell extract, and 4.82 ng/mL in human colon tissue extract.

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Product protocols

For this product, it's our understanding that no specific protocols are required. You can:

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