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AB257693

Human SNAP29 knockout HeLa cell lysate

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SNAP29 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 exon1 and 5 bp deletion in exon1.

View Alternative Names

Synaptosomal-associated protein 29, Vesicle-membrane fusion protein SNAP-29, SNAP29, SNAP-29, Soluble 29 kDa NSF attachment protein

4 Images
Western blot - Human SNAP29 knockout HeLa cell lysate (AB257693)
  • WB

Lab

Western blot - Human SNAP29 knockout HeLa cell lysate (AB257693)

Lane 1 : Wild-type HeLa cell lysate (20 ug)
Lane 2 : SNAP29 knockout HeLa cell lysate (20 ug)
Lane 3 : HepG2 cell lysate (20 ug)

ab181151 was shown to specifically react with SNAP29 in wild-type HeLa cells. Loss of signal was observed when knockout cell line ab265289 (knockout cell lysate ab257693) was used. Wild-type and SNAP29 knockout samples were subjected to SDS-PAGE. ab181151 and Anti-GAPDH antibody [6C5] - Loading Control (ab8245) were incubated overnight at 4oC 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-SNAP29 antibody [EPR9198(2)] (<a href='/en-us/products/primary-antibodies/snap29-antibody-epr91982-ab181151'>ab181151</a>) at 1/1000 dilution

Lane 1:

Wild-type HeLa cell lysate at 20 µg

Lane 2:

SNAP29 knockout HeLa cell lysate at 20 µg

Lane 2:

Western blot - Human SNAP29 knockout HeLa cell line (<a href='/en-us/products/cell-lines/human-snap29-knockout-hela-cell-line-ab265289'>ab265289</a>)

Lane 3:

HepG2 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: 29 kDa

Observed band size: 29 kDa

false

Western blot - Human SNAP29 knockout HeLa cell lysate (AB257693)
  • WB

Lab

Western blot - Human SNAP29 knockout HeLa cell lysate (AB257693)

Lane 1 : Wild-type HeLa cell lysate (20 ug)
Lane 2 : SNAP29 knockout HeLa cell lysate (20 ug)
Lane 3 : HepG2 cell lysate (20 ug)

ab138500 was shown to specifically react with SNAP29 in wild-type HeLa cells. Loss of signal was observed when knockout cell line ab265289 (knockout cell lysate ab257693) was used. Wild-type and SNAP29 knockout samples were subjected to SDS-PAGE. ab138500 and Anti-GAPDH antibody [6C5] - Loading Control (ab8245) were incubated overnight at 4oC 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-SNAP29 antibody [EPR9199] (<a href='/en-us/products/primary-antibodies/snap29-antibody-epr9199-ab138500'>ab138500</a>) at 1/1000 dilution

Lane 1:

Wild-type HeLa cell lysate at 20 µg

Lane 2:

SNAP29 knockout HeLa cell lysate at 20 µg

Lane 2:

Western blot - Human SNAP29 knockout HeLa cell line (<a href='/en-us/products/cell-lines/human-snap29-knockout-hela-cell-line-ab265289'>ab265289</a>)

Lane 3:

HepG2 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: 29 kDa

Observed band size: 29 kDa

false

Sanger Sequencing - Human SNAP29 knockout HeLa cell lysate (AB257693)
  • Sanger seq

Unknown

Sanger Sequencing - Human SNAP29 knockout HeLa cell lysate (AB257693)

Allele-2 : 1 bp insertion in exon1

Sanger Sequencing - Human SNAP29 knockout HeLa cell lysate (AB257693)
  • Sanger seq

Unknown

Sanger Sequencing - Human SNAP29 knockout HeLa cell lysate (AB257693)

Allele-1 : 5 bp deletion in exon1

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 exon1 and 5 bp deletion in exon1.

Disease

Adenocarcinoma

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Complementary Products

Primary Antibodies

AB181151

Anti-SNAP29 antibody [EPR9198(2)]

RabMAb|Recombinant|KO Validated

Immunocytochemistry/ Immunofluorescence - Anti-SNAP29 antibody [EPR9198(2)] (AB181151)

  • 5
  • 1
Primary Antibodies

AB36195

Anti-SYBL1/VAMP-7 antibody [158.2]

Flow Cytometry - Anti-SYBL1/VAMP-7 antibody [158.2] (AB36195)

  • 5
  • 1
Proteins & Peptides

AB259412

Recombinant human VEGF 165A protein (Active)

Premium Bioactive

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

  • 0
  • 0
Proteins & Peptides

AB288799

Recombinant Human VCAM1 protein (Active)

Premium Bioactive

Functional Studies - Recombinant Human VCAM1 protein (Active) (AB288799)

  • 0
  • 0
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Reactivity data

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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
SNAP29
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.

SNAP29 known also as synaptosomal-associated protein 29 functions mainly in membrane fusion. It weighs about 29 kDa. This protein plays a significant role in intracellular membrane trafficking guiding vesicle fusion events. It is expressed in many tissues with high occurrence in neuronal and non-neuronal cells. SNAP29 contains coiled-coil domains that facilitate its role in binding with SNARE complexes which assist in the docking and fusion of vesicles with target membranes.
Biological function summary

RNA encoding this protein facilitates vesicle-mediated transport on the cellular level especially under conditions requiring rapid membrane addition or reshaping. SNAP29 acts alongside other SNARE proteins like syntaxin and VAMP in the SNARE complex to modulate membrane interactions. The protein eases the process of cytoplasmic content exchange between organelles. As part of these complexes SNAP29 maintains cellular homeostasis by contributing to membrane dynamics and trafficking processes.

Pathways

Vesicular transport in the cell highly depends on SNAP29 especially in exocytosis and endocytosis pathways. Alongside other SNARE proteins SNAP29 fits into the vesicle docking and fusion pathways ensuring efficient membrane recycling and cargo delivery. It interacts with several proteins in the process including syntaxin 1 and VAMP2 to facilitate these important functions within these pathways.

Impairments or mutations in the SNAP29 gene link to conditions like CEDNIK syndrome and other neurodevelopmental disorders. CEDNIK syndrome arises from disrupted membrane trafficking and organelle biogenesis due to SNAP29 dysfunction. In these cases a relationship with other proteins in the disease pathways such as STXBP1 can also alter disease progression and severity influencing neuronal communication and stability.
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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'.

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