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AB257625

Human RAB9A (Rab9) knockout HeLa cell lysate

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RAB9A KO cell lysate available now. KO validated by Western blot. Free of charge wild type control included. Knockout achieved by using CRISPR/Cas9, 1 bp insertion in exon 3 and Insertion of the selection cassette in exon 3.
4 Images
Western blot - Human RAB9A (Rab9) knockout HeLa cell lysate (AB257625)
  • WB

Lab

Western blot - Human RAB9A (Rab9) knockout HeLa cell lysate (AB257625)

Lane 1 : Wild-type HeLa cell lysate (20 ug)
Lane 2 : RAB9A knockout HeLa cell lysate (20 ug)
Lane 3 : MCF7 cell lysate (20 ug)
Lane 4 : MDA-MB-231 cell lysate (20 ug)

ab179815 was shown to specifically react with Rab9 in wild-type HeLa cells. Loss of signal was observed when knockout cell line ab265693 (knockout cell lysate ab257625) was used. Wild-type and Rab9 knockout samples were subjected to SDS-PAGE. ab179815 and Anti-GAPDH antibody [6C5] - Loading Control (ab8245) were incubated at room temperature for 2.5 hours at 1 in 1000 dilution and 1 in 20000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye® 680RD) preadsorbed (ab216776) secondary antibodies at 1 in 20000 dilution for 1 hour at room temperature before imaging.

All lanes:

Western blot - Anti-Rab9 antibody [EPR13272] - Late Endosome Marker (<a href='/en-us/products/primary-antibodies/rab9-antibody-epr13272-late-endosome-marker-ab179815'>ab179815</a>) at 1/1000 dilution

Lane 1:

Wild-type HeLa cell lysate at 20 µg

Lane 2:

RAB9A knockout HeLa cell lysate at 20 µg

Lane 2:

Western blot - Human RAB9A (Rab9) knockout HeLa cell line (<a href='/en-us/products/cell-lines/human-rab9a-rab9-knockout-hela-cell-line-ab265693'>ab265693</a>)

Lane 3:

MCF7 cell lysate at 20 µg

Lane 4:

MDA-MB-231 cell lysate at 20 µg

Secondary

All lanes:

Western blot - Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed (<a href='/en-us/products/secondary-antibodies/goat-rabbit-igg-h-l-irdye-800cw-preadsorbed-ab216773'>ab216773</a>) at 1/10000 dilution

Predicted band size: 23 kDa

Observed band size: 25 kDa

false

Western blot - Human RAB9A (Rab9) knockout HeLa cell lysate (AB257625)
  • WB

Lab

Western blot - Human RAB9A (Rab9) knockout HeLa cell lysate (AB257625)

Lane 1 : Wild-type HeLa cell lysate (20 ug)
Lane 2 : RAB9A knockout HeLa cell lysate (20 ug)
Lane 3 : MCF7 cell lysate (20 ug)
Lane 4 : MDA-MB-231 cell lysate (20 ug)

ab2810 was shown to specifically react with Rab9 in wild-type HeLa cells. Loss of signal was observed when knockout cell line ab265693 (knockout cell lysate ab257625) was used. Wild-type and Rab9 knockout samples were subjected to SDS-PAGE. ab2810 and Anti-GAPDH antibody[EPR16891] - Loading Control (ab181602) were incubated at room temperature for 2.5 hours at 1 in 1000 dilution and 1 in 20000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye® 680RD) preadsorbed (ab216777) and Goat anti-Mouse IgG H&L (IRDye® 800CW) preadsorbed (ab216772) secondary antibodies at 1 in 20000 dilution for 1 hour at room temperature before imaging.

All lanes:

Western blot - Anti-Rab9 antibody [Mab9] - Late Endosome Marker (<a href='/en-us/products/primary-antibodies/rab9-antibody-mab9-late-endosome-marker-ab2810'>ab2810</a>) at 1/1000 dilution

Lane 1:

Wild-type HeLa cell lysate at 20 µg

Lane 2:

RAB9A knockout HeLa cell lysate at 20 µg

Lane 2:

Western blot - Human RAB9A (Rab9) knockout HeLa cell line (<a href='/en-us/products/cell-lines/human-rab9a-rab9-knockout-hela-cell-line-ab265693'>ab265693</a>)

Lane 3:

MCF7 cell lysate at 20 µg

Lane 4:

MDA-MB-231 cell lysate at 20 µg

Secondary

All lanes:

Western blot - Goat Anti-Rabbit IgG H&L (IRDye® 680RD) preadsorbed (<a href='/en-us/products/secondary-antibodies/goat-rabbit-igg-h-l-irdye-680rd-preadsorbed-ab216777'>ab216777</a>) at 1/10000 dilution

Predicted band size: 23 kDa

Observed band size: 25 kDa

false

Sanger Sequencing - Human RAB9A (Rab9) knockout HeLa cell lysate (AB257625)
  • Sanger seq

Unknown

Sanger Sequencing - Human RAB9A (Rab9) knockout HeLa cell lysate (AB257625)

Allele-1 : 1 bp insertion in exon 3

Sanger Sequencing - Human RAB9A (Rab9) knockout HeLa cell lysate (AB257625)
  • Sanger seq

Unknown

Sanger Sequencing - Human RAB9A (Rab9) knockout HeLa cell lysate (AB257625)

Allele-2 : Insertion of the selection cassette in exon 3

Key facts

Cell type

HeLa

Species or organism

Human

Tissue

Cervix

Knockout validation

Sanger Sequencing,Western blot

Mutation description

Knockout achieved by using CRISPR/Cas9, 1 bp insertion in exon 3 and Insertion of the selection cassette in exon 3.

Disease

Adenocarcinoma

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Complementary Products

Primary Antibodies

AB2810

Anti-Rab9 antibody [Mab9] - Late Endosome Marker

BOND RX™ Validated|KO Validated

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Rab9 antibody [Mab9] - Late Endosome Marker (AB2810)

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Proteins & Peptides

AB80157

Recombinant Human M6PR (cation dependent) protein

SDS-PAGE - Recombinant Human M6PR (cation dependent) protein (AB80157)

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Proteins & Peptides

AB259412

Recombinant human VEGF 165A protein (Active)

Premium Bioactive

Mass Spectrometry - Recombinant human VEGF 165A protein (Active) (AB259412)

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  • 0
Proteins & Peptides

AB180317

Recombinant Human POLR1C protein

SDS-PAGE - Recombinant Human POLR1C protein (AB180317)

  • 0
  • 0
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Reactivity data

{ "title": "Reactivity Data", "filters": { "stats": ["", "Reactivity", "Dilution Info", "Notes"] }, "values": { "WB": { "reactivity":"TESTED_AND_REACTS", "dilution-info":"", "notes":"<p></p>" } } }
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Product details

Knockout cell lysate achieved by CRISPR/Cas9.

REACH authorisation
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.

Lysate preparation: Our lysates are made using RIPA buffer to which we add a protease inhibitor cocktail and phosphatase inhibitor cocktail (ratio: 300:100:10). This means that the protein of interest is denatured. If you require a native form of the protein please use the live cell version. Please refer to our lysis protocol for further details on how our lysates are prepared.

User storage instructions: Lyophilizate may be stored at 4°C. After reconstitution, store at -20°C for short-term storage or -80°C for long-term storage.

This product is subject to limited use licenses from The Broad Institute, ERS Genomics Limited and Sigma-Aldrich Co. LLC, and is developed with patented technology. For full details of the licenses and patents please refer to our limited use license and patent pages.

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What's included?

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Properties and storage information

Gene name
RAB9A
Gene editing type
Knockout
Gene editing method
CRISPR technology
Knockout validation
Sanger Sequencing, Western blot
Shipped at conditions
Ambient - Can Ship with Ice
Appropriate short-term storage conditions
-20°C
Appropriate long-term storage conditions
-20°C
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Supplementary information

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

Rab9 also known as Ras-related protein Rab-9A is a small GTPase and part of the Rab family with a mass of around 23 kDa. This protein mainly resides in the trans-Golgi network and late endosomes. Rab9 plays an important role in the transport of mannose 6-phosphate receptors between late endosomes and the Golgi apparatus facilitating cargo sorting and vesicular trafficking in cells.
Biological function summary

Rab9 coordinates several intracellular trafficking processes aiding in the regulation of membrane identity. It associates with other effectors such as tail-interacting protein (TIP47) to form complexes on the endosomal surface. These complexes help in vesicular transport ensuring efficient recycling of receptors. Rab9 impacts endosome maturation and transport making it an essential endosome marker.

Pathways

Rab9 influences and is involved in the regulation of the endocytic and secretory pathways. It interacts with proteins like Rab7 another GTPase to manage late endosome and lysosome transport. These interactions underline Rab9's functionalities in intracellular trafficking and recycling processes relying on cooperative interactions with other Rabs and their effectors.

Rab9 plays a role in conditions like Niemann-Pick disease type C and some neurological disorders. Disruption in Rab9 function may affect lipid trafficking and lead to accumulation of cholesterol in late endosomes. This links Rab9 indirectly to proteins such as NPC1 which is known to be implicated in cholesterol homeostasis. Understanding Rab9's involvement can help inform therapeutic strategies targeting these pathways in related diseases.
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Quality control

STR analysis

CSF1PO, D13S317, D7S820, D5S818, TH01, D16S539, TPOX

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Cell culture

Biosafety level

EU: 2 US: 2

Adherent/suspension

Adherent

Gender

Female

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Product protocols

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Product promise

We are committed to supporting your work with high-quality reagents, and we're here for you every step of the way. In the unlikely event that one of our products does not perform as expected, you're protected by our Product Promise.
For full details, please see our Terms & Conditions

Please note: All products are 'FOR RESEARCH USE ONLY. NOT FOR USE IN DIAGNOSTIC OR THERAPEUTIC PROCEDURES'.

For licensing inquiries, please contact partnerships@abcam.com