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AB324111

Anti-PINK1 antibody [EPR29146-340] - BSA and Azide free

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Rabbit Recombinant Monoclonal PINK1 antibody. Carrier free. Suitable for I-ELISA, ICC/IF, WB and reacts with Recombinant fragment - Human, Human samples.

View Alternative Names

BRPK, PTEN-induced putative kinase protein 1, PINK1

5 Images
Immunocytochemistry/ Immunofluorescence - Anti-PINK1 antibody [EPR29146-340] - BSA and Azide free (AB324111)
  • ICC/IF

Supplier Data

Immunocytochemistry/ Immunofluorescence - Anti-PINK1 antibody [EPR29146-340] - BSA and Azide free (AB324111)

This data was developed using ab323807, the same antibody clone in a different buffer formulation.

Immunofluorescent analysis of 4% Paraformaldehyde-fixed 0.1% TritonX-100 permeabilized PINK1 KO HEK-293T (PINK1 knockout human embryonic kidney epithelial cell) ab266393 cells labelling PINK1 with ab323807 at 1/50 (10.34 ug/ml) dilution followed by ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed antibody at 1/1000 2 g/ml dilution (Green).

Confocal image showing increased mitochondrial staining in wildtype HEK-293T cells treated with 10 µM Valinomycin for 24 hours (shown in green). The counterstain was observed in magenta. Nuclear DNA was labelled with DAPI (shown in blue). Image was taken with a confocal microscope (Leica-Microsystems TCS SP8).

Secondary antibody only control : Secondary antibody is ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed at 1/1000 2 g/ml dilution.

anti-COX IV mouse monoclonal antibody - Mitochondrial Marker was used to counterstain tubulin at 1/200 2.5 dilution (Magenta). The Nuclear counterstain was DAPI (Blue).

Counterstain Secondary antibody only control : Secondary antibody is ab150120 Goat Anti-Mouse IgG H&L (Alexa Fluor® 594) preadsorbed at 1/1000 2 g/ml dilution.

Immunocytochemistry/ Immunofluorescence - Anti-PINK1 antibody [EPR29146-340] - BSA and Azide free (AB324111)
  • ICC/IF

Supplier Data

Immunocytochemistry/ Immunofluorescence - Anti-PINK1 antibody [EPR29146-340] - BSA and Azide free (AB324111)

This data was developed using ab323807, the same antibody clone in a different buffer formulation.

Immunofluorescent analysis of 4% Paraformaldehyde-fixed 0.1% TritonX-100 permeabilized SK-OV-3(human ovarian cancer epithelial cell) cells labelling PINK1 with ab323807 at 1/50 (10.34 ug/ml) dilution followed by ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed antibody at 1/1000 2 g/ml dilution (Green).

Confocal image showing mitochondrial staining in SK-OV-3 cells (shown in green) treatment with 30 µM CCCP for 6 hours. The counterstain was observed in magenta. Nuclear DNA was labelled with DAPI (shown in blue). Image was taken with a confocal microscope (Leica-Microsystems TCS SP8).

Secondary antibody only control : Secondary antibody is ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed at 1/1000 2 g/ml dilution.

anti-COX IV mouse monoclonal antibody - Mitochondrial Marker was used to counterstain tubulin at 1/200 2.5 dilution (Magenta). The Nuclear counterstain was DAPI (Blue).

Counterstain Secondary antibody only control : Secondary antibody is ab150120 Goat Anti-Mouse IgG H&L (Alexa Fluor® 594) preadsorbed at 1/1000 2 g/ml dilution.

Western blot - Anti-PINK1 antibody [EPR29146-340] - BSA and Azide free (AB324111)
  • WB

Supplier Data

Western blot - Anti-PINK1 antibody [EPR29146-340] - BSA and Azide free (AB324111)

This data was developed using ab323807, the same antibody clone in a different buffer formulation.

Blocking and diluting buffer and concentration : 5% NFDM/TBST.

Performed under reducing conditions.

In Western blot ab323807 was shown to bind specifically to PINK1. Target of interest was observed at 62 kDa in wild-type HEK-293 cell lysates (lane 2) with no signal observed at this size in PINK1 knockout cell line (lane 4) (lane 4 knockout cell line ab266393).

The identity of the higher MW band at approximately 250 kDa (in lanes 1-4) is unknown.

The expression of PINK1 is upregulated in response to CCCP treatment (PMID : 22724072).

In Western blot Anti-GAPDH antibody [EPR16891] - Loading Control (ab181602) staining at 1/200000 dilution.

All lanes:

Western blot - Anti-PINK1 antibody [EPR29146-340] (<a href='/en-us/products/primary-antibodies/pink1-antibody-epr29146-340-ab323807'>ab323807</a>) at 1/1000 dilution

Lane 1:

Untreated wild-type HEK-293 (human embryonic kidney epithelial cell) whole cell lysate at 80 µg

Lane 2:

Wild-type HEK-293 treated with 30μM CCCP for 6h, whole cell lysate at 80 µg

Lane 3:

Untreated PINK1 knockout HEK-293 whole cell lysate at 80 µg

Lane 4:

PINK1 knockout HEK-293 treated with 30μM CCCP for 6h, whole cell lysate at 80 µg

Secondary

All lanes:

Western blot - Goat Anti-Rabbit IgG H&L (HRP) (<a href='/en-us/products/secondary-antibodies/goat-rabbit-igg-h-l-hrp-ab97051'>ab97051</a>) at 1/20000 dilution

Observed band size: 62 kDa,36 kDa

false

Exposure time: 125s

Western blot - Anti-PINK1 antibody [EPR29146-340] - BSA and Azide free (AB324111)
  • WB

Supplier Data

Western blot - Anti-PINK1 antibody [EPR29146-340] - BSA and Azide free (AB324111)

This data was developed using ab323807, the same antibody clone in a different buffer formulation.

Blocking and diluting buffer and concentration : 5% NFDM/TBST.

Performed under reducing conditions.

In Western blot ab323807 was shown to bind specifically to PINK1. Target of interest was observed at 62 kDa in wild-type HEK-293 cell lysates (lane 2) with no signal observed at this size in PINK1 knockout cell line (lane 4) (lane 4 knockout cell line ab266393).

The identity of the higher MW band at approximately 250 kDa (in lanes 1-4) is unknown.

The expression of PINK1 is upregulated in response to valinomycin treatment (PMID : 22724072).

The expression of PINK1 is upregulated in response to CCCP treatment (PMID : 22724072).

In Western blot Anti-GAPDH antibody [EPR16891] - Loading Control (ab181602) staining at 1/200000 dilution.

Exposure time : Lanes 1-4 : 48 seconds Lanes 5-6 : 10 seconds

All lanes:

Western blot - Anti-PINK1 antibody [EPR29146-340] (<a href='/en-us/products/primary-antibodies/pink1-antibody-epr29146-340-ab323807'>ab323807</a>) at 1/1000 dilution

Lane 1:

Untreated wild-type HEK-293 (human embryonic kidney epithelial cell) whole cell lysate at 20 µg

Lane 2:

Wild-type HEK-293 treated with 10μM valinomycin for 24h, whole cell lysate at 20 µg

Lane 3:

Untreated PINK1 knockout HEK-293 whole cell lysate at 20 µg

Lane 4:

PINK1 knockout HEK-293 treated with 10μM valinomycin for 24h, whole cell lysate at 20 µg

Lane 5:

Untreated SK-OV-3 (human ovarian cancer epithelial cell) whole cell lysate at 20 µg

Lane 6:

SK-OV-3 treated with 30μM CCCP for 6h, whole cell lysate at 20 µg

Secondary

All lanes:

Western blot - Goat Anti-Rabbit IgG H&L (HRP) (<a href='/en-us/products/secondary-antibodies/goat-rabbit-igg-h-l-hrp-ab97051'>ab97051</a>) at 1/20000 dilution

Observed band size: 62 kDa,36 kDa

false

Indirect ELISA - Anti-PINK1 antibody [EPR29146-340] - BSA and Azide free (AB324111)
  • I-ELISA

Supplier Data

Indirect ELISA - Anti-PINK1 antibody [EPR29146-340] - BSA and Azide free (AB324111)

This data was developed using ab323807, the same antibody clone in a different buffer formulation.

Indirect ELISA analysis of ab323807 at 1000-0 ng/ml. The Secondary antibody used was Alkaline Phosphatase-conjugated AffiniPure Goat Anti-Rabbit IgG (H+L) at 1 : 2500 dilution dilution.

Antigen : Human PINK1.

Antigen concentration : 1000 ng/ml

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Key facts

Host species

Rabbit

Clonality

Monoclonal

Clone number

EPR29146-340

Isotype

IgG

Carrier free

Yes

Reacts with

Human

Applications

ICC/IF, WB, I-ELISA

applications

Immunogen

The exact immunogen used to generate this antibody is proprietary information.

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Reactivity data

{ "title": "Reactivity Data", "filters": { "stats": ["", "Species", "Dilution Info", "Notes"], "tabs": { "all-applications": {"fullname" : "All Applications", "shortname": "All Applications"}, "IELISA" : {"fullname" : "Indirect ELISA", "shortname":"I-ELISA"}, "ICCIF" : {"fullname" : "Immunocytochemistry/ Immunofluorescence", "shortname":"ICC/IF"}, "WB" : {"fullname" : "Western blot", "shortname":"WB"}, "IHCP" : {"fullname" : "Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)", "shortname":"IHC-P"} }, "product-promise": { "all": "all", "testedAndGuaranteed": "tested", "guaranteed": "expected", "predicted": "predicted", "notRecommended": "not-recommended" } }, "values": { "Human": { "IELISA-species-checked": "guaranteed", "IELISA-species-dilution-info": "", "IELISA-species-notes": "", "ICCIF-species-checked": "testedAndGuaranteed", "ICCIF-species-dilution-info": "", "ICCIF-species-notes": "<p></p>", "WB-species-checked": "testedAndGuaranteed", "WB-species-dilution-info": "", "WB-species-notes": "<p></p>", "IHCP-species-checked": "notRecommended", "IHCP-species-dilution-info": "", "IHCP-species-notes": "<p></p>" }, "Recombinant fragment - Human": { "IELISA-species-checked": "testedAndGuaranteed", "IELISA-species-dilution-info": "", "IELISA-species-notes": "<p></p>", "ICCIF-species-checked": "notRecommended", "ICCIF-species-dilution-info": "", "ICCIF-species-notes": "", "WB-species-checked": "notRecommended", "WB-species-dilution-info": "", "WB-species-notes": "", "IHCP-species-checked": "notRecommended", "IHCP-species-dilution-info": "", "IHCP-species-notes": "" } } }
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Product details

ab324111 is the carrier free version of ab323807.

Patented technology
Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMAb® patents.

What are the advantages of a recombinant monoclonal antibody?
This product is a recombinant monoclonal antibody, which offers several advantages including:

  • - High batch-to-batch consistency and reproducibility
  • - Improved sensitivity and specificity
  • - Long-term security of supply
  • - Animal-free batch production

For more information, read more on recombinant antibodies.

Conjugation ready
Our carrier-free antibodies are typically supplied in a PBS-only formulation, purified and free of BSA, sodium azide and glycerol. This conjugation-ready format is designed for use with fluorochromes, metal isotopes, oligonucleotides, and enzymes, which makes them ideal for antibody labelling, functional and cell-based assays, flow-based assays (e.g. mass cytometry) and Multiplex Imaging applications.

Use our conjugation kits for antibody conjugates that are ready-to-use in as little as 20 minutes with 1 minute hands-on-time and 100% antibody recovery: available for fluorescent dyes, HRP, biotin and gold.

Compatibility
This product is compatible with the Maxpar® Antibody Labeling Kit from Fluidigm, without the need for antibody preparation. Maxpar® is a trademark of Fluidigm Canada Inc.

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Properties and storage information

Form
Liquid
Purification technique
Affinity purification Protein A
Storage buffer
pH: 7.2 - 7.4 Constituents: PBS
Shipped at conditions
Blue Ice
Appropriate short-term storage conditions
+4°C
Appropriate long-term storage conditions
+4°C
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Product protocols

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

Serine/threonine-protein kinase which acts as a sensor of mitochondrial damage and protects against mitochondrial dysfunction during cellular stress (PubMed : 40080546). It phosphorylates mitochondrial proteins to coordinate mitochondrial quality control mechanisms that remove and replace dysfunctional mitochondrial components (PubMed : 14607334, PubMed : 15087508, PubMed : 18443288, PubMed : 18957282, PubMed : 19229105, PubMed : 19966284, PubMed : 20404107, PubMed : 20547144, PubMed : 20798600, PubMed : 22396657, PubMed : 23620051, PubMed : 23754282, PubMed : 23933751, PubMed : 24660806, PubMed : 24751536, PubMed : 24784582, PubMed : 24896179, PubMed : 24898855, PubMed : 25527291, PubMed : 32484300). In healthy mitochondria, PINK1 is translocated across the mitochondrial outer membrane (MOM) via the translocase of the outer membrane (TOM) complex, and inserted into the mitochondrial inner membrane (MIM) via the translocase of the inner membrane (TIM23) complex where it is cleaved and released into the cytosol (PubMed : 40080546). Depending on the severity of mitochondrial damage, activity ranges from preventing apoptosis and stimulating mitochondrial biogenesis to eliminating severely damaged mitochondria via PINK1-PRKN-dependent mitophagy (PubMed : 14607334, PubMed : 15087508, PubMed : 18443288, PubMed : 19966284, PubMed : 20404107, PubMed : 20798600, PubMed : 22396657, PubMed : 23620051, PubMed : 23933751, PubMed : 24898855, PubMed : 32047033, PubMed : 32484300). When cellular stress results in irreversible mitochondrial damage, PINK1 accumulates at the outer mitochondrial membrane (OMM) where it phosphorylates pre-existing polyubiquitin chains at 'Ser-65', recruits PRKN from the cytosol to the OMM and activates PRKN by phosphorylation at 'Ser-65'; activated PRKN then ubiquitinates VDAC1 and other OMM proteins to initiate mitophagy (PubMed : 14607334, PubMed : 15087508, PubMed : 19966284, PubMed : 20404107, PubMed : 20798600, PubMed : 23754282, PubMed : 23933751, PubMed : 24660806, PubMed : 24751536, PubMed : 24784582, PubMed : 25474007, PubMed : 25527291, PubMed : 32047033, PubMed : 40080546). The PINK1-PRKN pathway also promotes fission of damaged mitochondria through phosphorylation and PRKN-dependent degradation of mitochondrial proteins involved in fission such as MFN2 (PubMed : 18443288, PubMed : 23620051, PubMed : 24898855). This prevents the refusion of unhealthy mitochondria with the mitochondrial network or initiates mitochondrial fragmentation facilitating their later engulfment by autophagosomes (PubMed : 18443288, PubMed : 23620051). Also promotes mitochondrial fission independently of PRKN and ATG7-mediated mitophagy, via the phosphorylation and activation of DNM1L (PubMed : 18443288, PubMed : 32484300). Regulates motility of damaged mitochondria by promoting the ubiquitination and subsequent degradation of MIRO1 and MIRO2; in motor neurons, this likely inhibits mitochondrial intracellular anterograde transport along the axons which probably increases the chance of the mitochondria undergoing mitophagy in the soma (PubMed : 22396657). Required for ubiquinone reduction by mitochondrial complex I by mediating phosphorylation of complex I subunit NDUFA10 (By similarity). Phosphorylates LETM1, positively regulating its mitochondrial calcium transport activity (PubMed : 29123128).
See full target information PINK1
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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

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