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AB324849

Anti-Rat Poly PR antibody [PERA534G] - BSA and Azide free

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Rat Recombinant Monoclonal C9orf72 antibody. Carrier free. Suitable for WB, IHC-P, ICC/IF, I-ELISA and reacts with Transfected cell lysate - Human, Transfected cell line - Human, Synthetic peptide - Human samples.

View Alternative Names

DENND9, DENNL72, C9orf72, Guanine nucleotide exchange factor C9orf72

4 Images
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Rat Poly PR antibody [PERA534G] - BSA and Azide free (AB324849)
  • IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Rat Poly PR antibody [PERA534G] - BSA and Azide free (AB324849)

This data was developed using ab323367, the same antibody clone in a different buffer formulation.

Immunohistochemical analysis of paraffin-embedded Panel A HEK-293T (human embryonic kidney epithelial cell) transfected with a poly PR expression vector containing a EGFP-tag and Panel B HEK-293T transfected with empty vector containing a EGFP-tag labeling Poly PR with ab323367 at 1/5000 (0.217 ug/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Positive staining on (A) HEK-293T transfected with a poly PR expression vector, no staining on (B) HEK-293T transfected with empty vector. The section was incubated with ab323367 for 30 mins at room temperature.
The immunostaining was performed on a Leica Biosystems BOND® RX instrument

Counterstained with Hematoxylin.

Secondary antibody only control : Secondary antibody is a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Heat mediated antigen retrieval was performed with Tris-EDTA buffer (pH 9.0, Epitope Retrieval Solution2) for 20 mins

Immunocytochemistry/ Immunofluorescence - Anti-Rat Poly PR antibody [PERA534G] - BSA and Azide free (AB324849)
  • ICC/IF

Supplier Data

Immunocytochemistry/ Immunofluorescence - Anti-Rat Poly PR antibody [PERA534G] - BSA and Azide free (AB324849)

This data was developed using ab323367, the same antibody clone in a different buffer formulation.

Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% TritonX-100 permeabilized 293T (human embryonic kidney epithelial cell) cells labelling Poly PR with ab323367 at 1/500 (1.91 ug/ml) dilution, followed by ab150160 Goat Anti-Rat IgG H&L (Alexa Fluor® 594) antibody at 1/1000 2ug/ml dilution (Green).

Confocal image showing nuclear with cytoplasmic staining in 293T cells transfected with a poly PR expression vector containing a EGFP-tag®(shown in magenta). Nuclear DNA was labeled with DAPI (shown in blue).
Image was taken with a confocal microscope (Leica-Microsystems, TCS SP8).

Secondary antibody only control : Secondary antibody is ab150160 Goat Anti-Rat IgG H&L (Alexa Fluor® 594) at 1/1000 2ug/ml dilution.

Indirect ELISA - Anti-Rat Poly PR antibody [PERA534G] - BSA and Azide free (AB324849)
  • I-ELISA

Supplier Data

Indirect ELISA - Anti-Rat Poly PR antibody [PERA534G] - BSA and Azide free (AB324849)

This data was developed using ab323367, the same antibody clone in a different buffer formulation.

Indirect ELISA analysis of ab323367 at 2000-0 ng/ml. The Secondary antibody used was Alkaline Phosphatase-conjugated AffiniPure Goat Anti-Rat IgG (H+L) at 1 : 1000 dilution dilution.

Antigen : poly (PR)20 peptide.

Antigen concentration : 1000 ng/ml

Western blot - Anti-Rat Poly PR antibody [PERA534G] - BSA and Azide free (AB324849)
  • WB

Supplier Data

Western blot - Anti-Rat Poly PR antibody [PERA534G] - BSA and Azide free (AB324849)

This data was developed using ab323367, the same antibody clone in a different buffer formulation.

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

In Western blot, Anti-Vinculin antibody [EPR8185] (ab129002) staining at 1/10000 dilution.

In Western blot, Anti-GFP antibody [E385] (ab32146) staining at 35kDa dilution.

All lanes:

Western blot - Anti-Rat Poly PR antibody [PERA534G] (<a href='/en-us/products/primary-antibodies/rat-poly-pr-antibody-pera534g-ab323367'>ab323367</a>) at 1/1000 dilution

Lane 1:

293T (human embryonic kidney epithelial cell) transfected with an empty vector containi a EGPF-tag whole cell lysate at 20 µg

Lane 2:

293T cells transfected with a poly (PR)20 expression vector containi a EGFP-tag whole cell lysate at 20 µg

Secondary

All lanes:

Western blot - Goat Anti-Rat IgG H&L (HRP) (<a href='/en-us/products/secondary-antibodies/goat-rat-igg-h-l-hrp-ab205720'>ab205720</a>) at 1/20000 dilution

Observed band size: 35 kDa,124 kDa

false

Exposure time: 1s

Key facts

Host species

Rat

Clonality

Monoclonal

Clone number

PERA534G

Isotype

IgG2c

Carrier free

Yes

Reacts with

Human

Applications

I-ELISA, ICC/IF, IHC-P, WB

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"}, "WB" : {"fullname" : "Western blot", "shortname":"WB"}, "IHCP" : {"fullname" : "Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)", "shortname":"IHC-P"}, "ICCIF" : {"fullname" : "Immunocytochemistry/ Immunofluorescence", "shortname":"ICC/IF"}, "IELISA" : {"fullname" : "Indirect ELISA", "shortname":"I-ELISA"} }, "product-promise": { "all": "all", "testedAndGuaranteed": "tested", "guaranteed": "expected", "predicted": "predicted", "notRecommended": "not-recommended" } }, "values": { "Human": { "WB-species-checked": "predicted", "WB-species-dilution-info": "", "WB-species-notes": "", "IHCP-species-checked": "predicted", "IHCP-species-dilution-info": "", "IHCP-species-notes": "", "ICCIF-species-checked": "predicted", "ICCIF-species-dilution-info": "", "ICCIF-species-notes": "", "IELISA-species-checked": "predicted", "IELISA-species-dilution-info": "", "IELISA-species-notes": "" }, "Synthetic peptide - Human": { "WB-species-checked": "notRecommended", "WB-species-dilution-info": "", "WB-species-notes": "", "IHCP-species-checked": "notRecommended", "IHCP-species-dilution-info": "", "IHCP-species-notes": "", "ICCIF-species-checked": "notRecommended", "ICCIF-species-dilution-info": "", "ICCIF-species-notes": "", "IELISA-species-checked": "testedAndGuaranteed", "IELISA-species-dilution-info": "", "IELISA-species-notes": "<p></p>" }, "Transfected cell line - Human": { "WB-species-checked": "notRecommended", "WB-species-dilution-info": "", "WB-species-notes": "", "IHCP-species-checked": "testedAndGuaranteed", "IHCP-species-dilution-info": "", "IHCP-species-notes": "<p></p> Perform heat-mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.", "ICCIF-species-checked": "testedAndGuaranteed", "ICCIF-species-dilution-info": "", "ICCIF-species-notes": "<p></p>", "IELISA-species-checked": "notRecommended", "IELISA-species-dilution-info": "", "IELISA-species-notes": "" }, "Transfected cell lysate - Human": { "WB-species-checked": "testedAndGuaranteed", "WB-species-dilution-info": "", "WB-species-notes": "<p></p>", "IHCP-species-checked": "notRecommended", "IHCP-species-dilution-info": "", "IHCP-species-notes": "", "ICCIF-species-checked": "notRecommended", "ICCIF-species-dilution-info": "", "ICCIF-species-notes": "", "IELISA-species-checked": "notRecommended", "IELISA-species-dilution-info": "", "IELISA-species-notes": "" } } }
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Product details

ab324849 is the carrier-free version of ab323367.

Want a custom formulation?
This antibody clone is manufactured by Abcam. If you require a custom buffer formulation or conjugation for your experiments, please contact orders@abcam.com

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

Component of the C9orf72-SMCR8 complex, a complex that has guanine nucleotide exchange factor (GEF) activity and regulates autophagy (PubMed : 27103069, PubMed : 27193190, PubMed : 27617292, PubMed : 28195531, PubMed : 32303654). In the complex, C9orf72 and SMCR8 probably constitute the catalytic subunits that promote the exchange of GDP to GTP, converting inactive GDP-bound RAB8A and RAB39B into their active GTP-bound form, thereby promoting autophagosome maturation (PubMed : 27103069). The C9orf72-SMCR8 complex also acts as a regulator of autophagy initiation by interacting with the ULK1/ATG1 kinase complex and modulating its protein kinase activity (PubMed : 27617292). As part of the C9orf72-SMCR8 complex, stimulates RAB8A and RAB11A GTPase activity in vitro (PubMed : 32303654). Positively regulates initiation of autophagy by regulating the RAB1A-dependent trafficking of the ULK1/ATG1 kinase complex to the phagophore which leads to autophagosome formation (PubMed : 27334615). Acts as a regulator of mTORC1 signaling by promoting phosphorylation of mTORC1 substrates (PubMed : 27559131). Plays a role in endosomal trafficking (PubMed : 24549040). May be involved in regulating the maturation of phagosomes to lysosomes (By similarity). Promotes the lysosomal localization and lysosome-mediated degradation of CARM1 which leads to inhibition of starvation-induced lipid metabolism (By similarity). Regulates actin dynamics in motor neurons by inhibiting the GTP-binding activity of ARF6, leading to ARF6 inactivation (PubMed : 27723745). This reduces the activity of the LIMK1 and LIMK2 kinases which are responsible for phosphorylation and inactivation of cofilin, leading to CFL1/cofilin activation (PubMed : 27723745). Positively regulates axon extension and axon growth cone size in spinal motor neurons (PubMed : 27723745). Required for SMCR8 protein expression and localization at pre- and post-synaptic compartments in the forebrain, also regulates protein abundance of RAB3A and GRIA1/GLUR1 in post-synaptic compartments in the forebrain and hippocampus (By similarity). Plays a role within the hematopoietic system in restricting inflammation and the development of autoimmunity (By similarity).. Isoform 1. Regulates stress granule assembly in response to cellular stress.. Isoform 2. Does not play a role in regulation of stress granule assembly in response to cellular stress.
See full target information C9orf72
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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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