AB322354

Anti-FAK (phospho Y397 + Y576 + Y925) antibody [RM1141]

20ul selling size
RabMAb
Recombinant
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Anti-FAK (phospho Y397 + Y576 + Y925) antibody [RM1141] is a Rabbit Recombinant Multiclonal antibody that is used in FAK (phospho Y397 + Y576 + Y925) Dot Blot, Western Blot. Suitable for Human, Mouse, Rat samples.

FAK (PTK2) is a non-receptor protein-tyrosine kinase which is involved in regulation of a number of essential cellular processes, including cell cycle progression, cell migration, actic cytoskeleton organisation and formation of focal adhesions. FAK is activated by phosphorylation and recruited to focal adhesions in response to integrin signalling. Aberrant expression of FAK may play a role in tumor formation, including ovarian, breast and non-small cell lung cancers.

4 Images

Supplier Data

Western blot - Anti-FAK (phospho Y397 + Y576 + Y925) antibody [RM1141] (AB322354)

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

The expression of phosphorylated FAK is upregulated in response to pervanadate treatment (PMID : 20847951).

In Western blot, Anti-Histone H3 antibody [EPR16987] - Nuclear Marker and ChIP Grade (ab176842) staining at 1/100000 dilution.

In Western blot, Anti-FAK antibody [EP695Y] (ab40794) staining at 1/1000 dilution.

All lanes:

Western blot - Anti-FAK (phospho Y397 + Y576 + Y925) antibody [RM1141] (ab322354) at 1/1000 dilution

Lane 1:

Untreated HeLa (human cervical adenocarcinoma epithelial cell) whole cell lysate (untreated membrane) at 20 µg

Lane 2:

HeLa treated with 10μM pervanadate for 60 minutes whole cell lysate (untreated membrane) at 20 µg

Lane 3:

Untreated HeLa whole cell lysate (alkaline phosphatase treated membrane) at 20 µg

Lane 4:

HeLa treated with 10μM pervanadate for 60 minutes whole cell lysate (alkaline phosphatase treated membrane) 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 1/100000 dilution

Observed band size: 119 kDa,15 kDa

false

Exposure time: 8s

Supplier Data

Dot Blot - Anti-FAK (phospho Y397 + Y576 + Y925) antibody [RM1141] (AB322354)

Dot blot analysis of FAK (phospho Y397 + Y576 + Y925) using ab322354 at 1 : 1000 (0.515 ug/ml) followed by a Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated (ab97051) at 1 : 100,000 dilution.

Lane1 : Human FAK (phospho Y397) peptide
Lane2 : Human FAK non-phospho peptide
Lane3 : Human FAK (phospho Y576) peptide
Lane4 : Human FAK non-phospho peptide
Lane5 : Human FAK (phospho Y925) peptide
Lane6 : Human FAK non-phospho peptide

Exposure time : 180 seconds.

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

All lanes:

Dot Blot - Anti-FAK (phospho Y397 + Y576 + Y925) antibody [RM1141] (ab322354) at 1/1000 dilution

Lane 1:

Human FAK (phospho Y397) peptide

Lanes 2, 4 and 6:

Human FAK non-phospho peptide

Lane 3:

Human FAK (phospho Y576) peptide

Lane 5:

Human FAK (phospho Y925) peptide

Secondary

All lanes:

Dot 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 1/100000 dilution

false

Exposure time: 180s

Supplier Data

Western blot - Anti-FAK (phospho Y397 + Y576 + Y925) antibody [RM1141] (AB322354)

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

The expression of phosphorylated FAK is upregulated in response to pervanadate treatment (PMID : 20847951).

In Western blot, Anti-Histone H3 antibody [EPR16987] - Nuclear Marker and ChIP Grade (ab176842) staining at 1/100000 dilution.

In Western blot, Anti-FAK antibody [EP695Y] (ab40794) staining at 1/1000 dilution.

All lanes:

Western blot - Anti-FAK (phospho Y397 + Y576 + Y925) antibody [RM1141] (ab322354) at 1/1000 dilution

Lane 1:

Untreated NIH/3T3 (mouse embryonic fibroblast) whole cell lysate (untreated membrane) at 20 µg

Lane 2:

NIH/3T3 treated with 10μM pervanadate for 60 minutes whole cell lysate (untreated membrane) at 20 µg

Lane 3:

Untreated NIH/3T3 whole cell lysate (alkaline phosphatase treated membrane) at 20 µg

Lane 4:

NIH/3T3 treated with 10μM pervanadate for 60 minutes whole cell lysate (alkaline phosphatase treated membrane) 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 1/100000 dilution

Observed band size: 119 kDa,15 kDa

false

Exposure time: 8s

Supplier Data

Western blot - Anti-FAK (phospho Y397 + Y576 + Y925) antibody [RM1141] (AB322354)

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

The activation of phosphorylation of FAK is reported to be related to developmental processes in brain tissue (PMID : 14642275). So expression level of phosphorylated modified FAK in normal brain is quite low causing not easy to be detected. We suggest optimizing experimental protocols (increasing lysate amount, using lower dilution or higher-sensitivity ECL substrate) to improve results.

FAK can be cleaved at several sites. The molecular weight observed is consistent with what has been described in the literature (PMID : 10545505, PMID : 9642276).

This blot of Lane 2 and 4 was developed using a high-sensitivity ECL substrate, allowing for the detection of proteins in the mid-femtogram range.

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

In Western blot, Anti-FAK antibody [EP695Y] (ab40794) staining at 1/1000 dilution.

Exposure time : Lane 1, 3 : 136 seconds; Lane 2, 4 : 70 seconds

All lanes:

Western blot - Anti-FAK (phospho Y397 + Y576 + Y925) antibody [RM1141] (ab322354) at 1/1000 dilution

Lane 1:

Mouse brain tissue lysate (untreated membrane) at 40 µg

Lane 2:

Rat brain tissue lysate (untreated membrane) at 40 µg

Lane 3:

Mouse brain tissue lysate (alkaline phosphatase treated membrane) at 40 µg

Lane 4:

Rat brain tissue lysate (alkaline phosphatase treated membrane) at 40 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 1/100000 dilution

Observed band size: 119 kDa,36 kDa

false

Key facts

Host species

Rabbit

Clonality

Multiclonal

Clone number

RM1141

Isotype

IgG

Carrier free

Reacts with

Human, Mouse, Rat

Applications

WB, Dot

applications

Immunogen

This product was produced with the following immunogens:

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"}, "Dot" : {"fullname" : "Dot Blot", "shortname":"Dot"}, "WB" : {"fullname" : "Western blot", "shortname":"WB"}, "IHCP" : {"fullname" : "Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)", "shortname":"IHC-P"}, "IP" : {"fullname" : "Immunoprecipitation", "shortname":"IP"} }, "product-promise": { "all": "all", "testedAndGuaranteed": "tested", "guaranteed": "expected", "predicted": "predicted", "notRecommended": "not-recommended" } }, "values": { "Human": { "Dot-species-checked": "guaranteed", "Dot-species-dilution-info": "", "Dot-species-notes": "", "WB-species-checked": "testedAndGuaranteed", "WB-species-dilution-info": "1/1000", "WB-species-notes": "", "IHCP-species-checked": "notRecommended", "IHCP-species-dilution-info": "", "IHCP-species-notes": "", "IP-species-checked": "notRecommended", "IP-species-dilution-info": "", "IP-species-notes": "" }, "Mouse": { "Dot-species-checked": "guaranteed", "Dot-species-dilution-info": "", "Dot-species-notes": "", "WB-species-checked": "testedAndGuaranteed", "WB-species-dilution-info": "1/1000", "WB-species-notes": "", "IHCP-species-checked": "notRecommended", "IHCP-species-dilution-info": "", "IHCP-species-notes": "", "IP-species-checked": "notRecommended", "IP-species-dilution-info": "", "IP-species-notes": "" }, "Rat": { "Dot-species-checked": "guaranteed", "Dot-species-dilution-info": "", "Dot-species-notes": "", "WB-species-checked": "testedAndGuaranteed", "WB-species-dilution-info": "1/1000", "WB-species-notes": "", "IHCP-species-checked": "notRecommended", "IHCP-species-dilution-info": "", "IHCP-species-notes": "", "IP-species-checked": "notRecommended", "IP-species-dilution-info": "", "IP-species-notes": "" }, "Synthetic peptide - Human": { "Dot-species-checked": "testedAndGuaranteed", "Dot-species-dilution-info": "1/1000", "Dot-species-notes": "", "WB-species-checked": "notRecommended", "WB-species-dilution-info": "", "WB-species-notes": "", "IHCP-species-checked": "notRecommended", "IHCP-species-dilution-info": "", "IHCP-species-notes": "", "IP-species-checked": "notRecommended", "IP-species-dilution-info": "", "IP-species-notes": "" } } }

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Product details

What are recombinant multiclonals?
Recombinant multiclonals are a mixture of recombinant antibodies co-expressed from a library of heavy and light chains. They offer several advantages including:

- The sensitivity of polyclonal antibodies by recognising multiple epitopes
- High batch-to-batch consistency and reproducibility
- Improved sensitivity and specificity
- Long-term security of supply
- Animal-free batch production

View our range of recombinant multiclonal antibodies.

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.

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Properties and storage information

Form

Liquid

Purification technique

Affinity purification Protein A

Storage buffer

pH: 7.2 - 7.4 Preservative: 0.01% Sodium azide Constituents: PBS, 40% Glycerol (glycerin, glycerine), 0.05% BSA

Shipped at conditions

Blue Ice

Appropriate short-term storage duration

1-2 weeks

Appropriate short-term storage conditions

+4°C

Appropriate long-term storage conditions

-20°C

Aliquoting information

Upon delivery aliquot

Storage information

Avoid freeze / thaw cycle

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Product protocols

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

Visit the General protocols
Visit the Troubleshooting

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Target data

Non-receptor protein-tyrosine kinase that plays an essential role in regulating cell migration, adhesion, spreading, reorganization of the actin cytoskeleton, formation and disassembly of focal adhesions and cell protrusions, cell cycle progression, cell proliferation and apoptosis. Required for early embryonic development and placenta development. Required for embryonic angiogenesis, normal cardiomyocyte migration and proliferation, and normal heart development. Regulates axon growth and neuronal cell migration, axon branching and synapse formation; required for normal development of the nervous system. Plays a role in osteogenesis and differentiation of osteoblasts. Functions in integrin signal transduction, but also in signaling downstream of numerous growth factor receptors, G-protein coupled receptors (GPCR), EPHA2, netrin receptors and LDL receptors. Forms multisubunit signaling complexes with SRC and SRC family members upon activation; this leads to the phosphorylation of additional tyrosine residues, creating binding sites for scaffold proteins, effectors and substrates. Regulates numerous signaling pathways. Promotes activation of phosphatidylinositol 3-kinase and the AKT1 signaling cascade. Promotes activation of MAPK1/ERK2, MAPK3/ERK1 and the MAP kinase signaling cascade. Promotes localized and transient activation of guanine nucleotide exchange factors (GEFs) and GTPase-activating proteins (GAPs), and thereby modulates the activity of Rho family GTPases. Signaling via CAS family members mediates activation of RAC1. Phosphorylates NEDD9 following integrin stimulation (PubMed : 9360983). Recruits the ubiquitin ligase MDM2 to P53/TP53 in the nucleus, and thereby regulates P53/TP53 activity, P53/TP53 ubiquitination and proteasomal degradation. Phosphorylates SRC; this increases SRC kinase activity. Phosphorylates ACTN1, ARHGEF7, GRB7, RET and WASL. Promotes phosphorylation of PXN and STAT1; most likely PXN and STAT1 are phosphorylated by a SRC family kinase that is recruited to autophosphorylated PTK2/FAK1, rather than by PTK2/FAK1 itself. Promotes phosphorylation of BCAR1; GIT2 and SHC1; this requires both SRC and PTK2/FAK1. Promotes phosphorylation of BMX and PIK3R1. Isoform 6 (FRNK) does not contain a kinase domain and inhibits PTK2/FAK1 phosphorylation and signaling. Its enhanced expression can attenuate the nuclear accumulation of LPXN and limit its ability to enhance serum response factor (SRF)-dependent gene transcription.. Isoform 6. Isoform 6 (FRNK) does not contain a kinase domain and inhibits PTK2/FAK1 phosphorylation and signaling. Its enhanced expression can attenuate the nuclear accumulation of LPXN and limit its ability to enhance serum response factor (SRF)-dependent gene transcription.

See full target information PTK2 pY397 + Y576 + Y925

Additional targets

Ptk2 phospho Y397 + Y576 + Y925,Ptk2 phospho Y397 + Y576 + Y925

Alternative Names

FAK, FAK1, Focal adhesion kinase 1, FADK 1, Focal adhesion kinase-related nonkinase, Protein phosphatase 1 regulatory subunit 71, Protein-tyrosine kinase 2, p125FAK, pp125FAK, FRNK, PPP1R71, Fak, Fak1, Focal adhesion kinase 1, FADK 1, Focal adhesion kinase-related nonkinase, Protein-tyrosine kinase 2, p125FAK, pp125FAK, FRNK, Fadk, Fak, Fak1, Kiaa4203, Focal adhesion kinase 1, FADK 1, Focal adhesion kinase-related nonkinase, Protein-tyrosine kinase 2, p125FAK, pp125FAK, FRNK

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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

Anti-FAK (phospho Y397 + Y576 + Y925) antibody [RM1141]

Western blot - Anti-FAK (phospho Y397 + Y576 + Y925) antibody [RM1141] (AB322354)

Dot Blot - Anti-FAK (phospho Y397 + Y576 + Y925) antibody [RM1141] (AB322354)

Western blot - Anti-FAK (phospho Y397 + Y576 + Y925) antibody [RM1141] (AB322354)

Western blot - Anti-FAK (phospho Y397 + Y576 + Y925) antibody [RM1141] (AB322354)

Key facts

Host species

Clonality

Clone number

Isotype

Carrier free

Reacts with

Applications

Immunogen

Complementary Products

Primary Antibodies

Primary Antibodies

Primary Antibodies

Primary Antibodies

Reactivity data

Product details

Properties and storage information

Form

Purification technique

Storage buffer

Shipped at conditions

Appropriate short-term storage duration

Appropriate short-term storage conditions

Appropriate long-term storage conditions

Aliquoting information

Storage information

Product protocols

Target data

Additional targets

Alternative Names

Product promise