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AB324785

Anti-CDK4 antibody [EPR28092-70] - BSA and Azide free

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Rabbit Recombinant Monoclonal CDK4 antibody. Carrier free. Suitable for Dot, IHC-P, WB and reacts with Transfected cell lysate, Mouse, Rat samples.

View Alternative Names

Crk3, Cdk4, Cyclin-dependent kinase 4, CRK3, Cell division protein kinase 4, PSK-J3

8 Images
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CDK4 antibody [EPR28092-70] - BSA and Azide free (AB324785)
  • IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CDK4 antibody [EPR28092-70] - BSA and Azide free (AB324785)

This data was developed using ab322262, the same antibody clone in a different buffer formulation.

Immunohistochemical analysis of paraffin-embedded Mouse breast cancer tissue labeling CDK4 with ab322262 at 1/4000 (0.134 ug/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Positive staining on mouse breast cancer.
The section was incubated with ab322262 for 30 mins at room temperature.
The immunostaining was performed on a Leica Biosystems BOND® RX instrument
Slides were incubated with 3% Hydrogen Peroxide for 10 mins at room temperature after secondary antibody incubation to reduce the background.

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

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CDK4 antibody [EPR28092-70] - BSA and Azide free (AB324785)
  • IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CDK4 antibody [EPR28092-70] - BSA and Azide free (AB324785)

This data was developed using ab322262, the same antibody clone in a different buffer formulation.

Immunohistochemical analysis of paraffin-embedded Mouse glioblastoma tissue labeling CDK4 with ab322262 at 1/4000 (0.134 ug/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Positive staining on mouse glioblastoma.
The section was incubated with ab322262 for 30 mins at room temperature.
The immunostaining was performed on a Leica Biosystems BOND® RX instrument
Slides were incubated with 3% Hydrogen Peroxide for 10 mins at room temperature after secondary antibody incubation to reduce the background.

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

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CDK4 antibody [EPR28092-70] - BSA and Azide free (AB324785)
  • IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CDK4 antibody [EPR28092-70] - BSA and Azide free (AB324785)

This data was developed using ab322262, the same antibody clone in a different buffer formulation.

Immunohistochemical analysis of paraffin-embedded Mouse ovary tissue labeling CDK4 with ab322262 at 1/4000 (0.134 ug/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Positive staining on mouse ovary.
The section was incubated with ab322262 for 30 mins at room temperature.
The immunostaining was performed on a Leica Biosystems BOND® RX instrument
Slides were incubated with 3% Hydrogen Peroxide for 10 mins at room temperature after secondary antibody incubation to reduce the background.

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

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CDK4 antibody [EPR28092-70] - BSA and Azide free (AB324785)
  • IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CDK4 antibody [EPR28092-70] - BSA and Azide free (AB324785)

This data was developed using ab322262, the same antibody clone in a different buffer formulation.

Immunohistochemical analysis of paraffin-embedded Mouse squamous cell carcinoma tissue labeling CDK4 with ab322262 at 1/4000 (0.134 ug/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Positive staining on mouse squamous cell carcinoma.
The section was incubated with ab322262 for 30 mins at room temperature.
The immunostaining was performed on a Leica Biosystems BOND® RX instrument
Slides were incubated with 3% Hydrogen Peroxide for 10 mins at room temperature after secondary antibody incubation to reduce the background.

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

Western blot - Anti-CDK4 antibody [EPR28092-70] - BSA and Azide free (AB324785)
  • WB

Supplier Data

Western blot - Anti-CDK4 antibody [EPR28092-70] - BSA and Azide free (AB324785)

This data was developed using ab322262, the same antibody clone in a different buffer formulation.

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

The expression profile/molecular weight observed is consistent with what has been described in the reference (PMID : 24091631), This antibody can recognize multiple isoforms of CDK4 and isoforms have different molecular weight.

Exposure time : Lanes 1-3 : 15 seconds; Lanes 4-5 : 114 seconds

All lanes:

Western blot - Anti-CDK4 antibody [EPR28092-70] (<a href='/en-us/products/primary-antibodies/cdk4-antibody-epr28092-70-ab322262'>ab322262</a>) at 1/1000 dilution

Lane 1:

C2C12 (mouse myoblast) whole cell lysate at 20 µg

Lane 2:

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

Lane 3:

4T1 (mouse mammary gland carcinoma epithelial cell) whole cell lysate at 20 µg

Lane 4:

C6 (rat glial tumor glial cell) whole cell lysate at 20 µg

Lane 5:

PC-12 (rat adrenal gland pheochromocytoma cell) 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: 33 kDa,30 kDa

false

Western blot - Anti-CDK4 antibody [EPR28092-70] - BSA and Azide free (AB324785)
  • WB

Supplier Data

Western blot - Anti-CDK4 antibody [EPR28092-70] - BSA and Azide free (AB324785)

This data was developed using ab322262, the same antibody clone in a different buffer formulation.

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

Low expression : brain (PMID : 30763280), skeletal muscle.

The expression profile/molecular weight observed is consistent with what has been described in the reference (PMID : 24091631), This antibody can recognize multiple isoforms of CDK4 and isoforms have different molecular weight.

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

Exposure time : Lanes 1-4 : 37 seconds; Lanes 5-7 : 114 seconds

All lanes:

Western blot - Anti-CDK4 antibody [EPR28092-70] (<a href='/en-us/products/primary-antibodies/cdk4-antibody-epr28092-70-ab322262'>ab322262</a>) at 1/1000 dilution

Lane 1:

Mouse ovary tissue lysate at 20 µg

Lane 2:

Mouse testis tissue lysate at 20 µg

Lane 3:

Mouse brain tissue lysate at 20 µg

Lane 4:

Mouse skeletal muscle tissue lysate at 20 µg

Lane 5:

Rat testis tissue lysate at 20 µg

Lane 6:

Rat ovary tissue lysate at 20 µg

Lane 7:

Rat skeletal muscle tissue 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: 33 kDa,30 kDa,36 kDa

false

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CDK4 antibody [EPR28092-70] - BSA and Azide free (AB324785)
  • IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CDK4 antibody [EPR28092-70] - BSA and Azide free (AB324785)

This data was developed using ab322262, the same antibody clone in a different buffer formulation.

Immunohistochemical analysis of paraffin-embedded Mouse skeletal muscle tissue labeling CDK4 with ab322262 at 1/4000 (0.134 ug/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Low expression tissue : No staining on mouse skeletal muscle.
The section was incubated with ab322262 for 30 mins at room temperature.
The immunostaining was performed on a Leica Biosystems BOND® RX instrument
Slides were incubated with 3% Hydrogen Peroxide for 10 mins at room temperature after secondary antibody incubation to reduce the background.

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

Dot Blot - Anti-CDK4 antibody [EPR28092-70] - BSA and Azide free (AB324785)
  • Dot

Supplier Data

Dot Blot - Anti-CDK4 antibody [EPR28092-70] - BSA and Azide free (AB324785)

This data was developed using ab322262, the same antibody clone in a different buffer formulation.

Dot blot analysis of CDK4 using ab322262 at 1 : 1000 (0.536 ug/ml) followed by a Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated (ab97051) at 1 : 20,000 dilution.

Lane1 : Purified 293T cells transfected with a mouse CDK4 expression vector containing a His-tag, whole cell lysate
Lane2 : Purified 293T cells transfected with a mouse CDK2 expression vector containing a His-tag, whole cell lysate
Lane3 : Purified 293T cells transfected with a mouse CDK6 expression vector containing a His-tag, whole cell lysate

Exposure time : 180 seconds.

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

This antibody does not cross-react with mouse CDK2 and CDK6.

In Dot Blot, Anti-6X His tag® antibody [EPR20547] - ChIP Grade (ab213204) staining at 1/5000 dilution.

All lanes:

Dot Blot - Anti-CDK4 antibody [EPR28092-70] (<a href='/en-us/products/primary-antibodies/cdk4-antibody-epr28092-70-ab322262'>ab322262</a>) at 1/1000 dilution

Lane 1:

Purified 293T cells transfected with a mouse CDK4 expression vector containing a His-tag, whole cell lysate

Lane 2:

Purified 293T cells transfected with a mouse CDK2 expression vector containing a His-tag, whole cell lysate

Lane 3:

Purified 293T cells transfected with a mouse CDK6 expression vector containing a His-tag, whole cell lysate

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 1/20000 dilution

false

Exposure time: 180s

Key facts

Host species

Rabbit

Clonality

Monoclonal

Clone number

EPR28092-70

Isotype

IgG

Carrier free

Yes

Reacts with

Mouse, Rat

Applications

IHC-P, WB, Dot

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"}, "Dot" : {"fullname" : "Dot Blot", "shortname":"Dot"}, "IHCP" : {"fullname" : "Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)", "shortname":"IHC-P"}, "WB" : {"fullname" : "Western blot", "shortname":"WB"} }, "product-promise": { "all": "all", "testedAndGuaranteed": "tested", "guaranteed": "expected", "predicted": "predicted", "notRecommended": "not-recommended" } }, "values": { "Human": { "Dot-species-checked": "notRecommended", "Dot-species-dilution-info": "", "Dot-species-notes": "", "IHCP-species-checked": "notRecommended", "IHCP-species-dilution-info": "", "IHCP-species-notes": "", "WB-species-checked": "notRecommended", "WB-species-dilution-info": "", "WB-species-notes": "<p></p>" }, "Mouse": { "Dot-species-checked": "guaranteed", "Dot-species-dilution-info": "", "Dot-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.", "WB-species-checked": "testedAndGuaranteed", "WB-species-dilution-info": "", "WB-species-notes": "<p></p>" }, "Rat": { "Dot-species-checked": "guaranteed", "Dot-species-dilution-info": "", "Dot-species-notes": "", "IHCP-species-checked": "guaranteed", "IHCP-species-dilution-info": "", "IHCP-species-notes": "", "WB-species-checked": "testedAndGuaranteed", "WB-species-dilution-info": "", "WB-species-notes": "<p></p>" }, "Transfected cell lysate": { "Dot-species-checked": "testedAndGuaranteed", "Dot-species-dilution-info": "", "Dot-species-notes": "<p></p>", "IHCP-species-checked": "notRecommended", "IHCP-species-dilution-info": "", "IHCP-species-notes": "", "WB-species-checked": "notRecommended", "WB-species-dilution-info": "", "WB-species-notes": "" } } }
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Product details

ab324785 is the carrier-free version of ab322262.

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: 100% 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

Ser/Thr-kinase component of cyclin D-CDK4 (DC) complexes that phosphorylate and inhibit members of the retinoblastoma (RB) protein family including RB1 and regulate the cell-cycle during G(1)/S transition. Phosphorylation of RB1 allows dissociation of the transcription factor E2F from the RB/E2F complexes and the subsequent transcription of E2F target genes which are responsible for the progression through the G(1) phase. Hypophosphorylates RB1 in early G(1) phase. Cyclin D-CDK4 complexes are major integrators of various mitogenenic and antimitogenic signals. Also phosphorylates SMAD3 in a cell-cycle-dependent manner and represses its transcriptional activity. Component of the ternary complex, cyclin D/CDK4/CDKN1B, required for nuclear translocation and activity of the cyclin D-CDK4 complex (By similarity).
See full target information Cdk4

Additional targets

Cdk4
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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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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