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AB302505

Anti-BCKDHA (phospho S293) antibody [EPR27489-76] - BSA and Azide free

  • BOND RX™ Validated
  • RabMAb
  • Recombinant
  • KO Validated
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Rabbit Recombinant Monoclonal BCKDHA phospho S293 antibody. Carrier free. Suitable for IP, IHC-P, WB and reacts with Mouse, Human, Rat samples.

View Alternative Names

Branched-chain alpha-keto acid dehydrogenase E1 component alpha chain, BCKDE1A, BCKDH E1-alpha, BCKDHA

7 Images
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-BCKDHA (phospho S293) antibody [EPR27489-76] - BSA and Azide free (AB302505)
  • IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-BCKDHA (phospho S293) antibody [EPR27489-76] - BSA and Azide free (AB302505)

This data was developed using ab302504, the same antibody clone in a different buffer formulation. Immunohistochemical analysis of paraffin-embedded Panel A. Human colon tissue labeling BCKDHA (phospho S293) with ab302504 at 1/500 (0.948 ug/ml) followed by a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101). Mitochondrial staining on human colon cancer without alkaline phosphatase treatment (Panel A). No signal was detected when tissues were treated with alkaline phosphatase (Panel B) (PMID : 32238881). The section was incubated with ab302504 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 Rabbit specific IHC polymer detection kit HRP/DAB (ab209101). Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution2) for 20 mins

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-BCKDHA (phospho S293) antibody [EPR27489-76] - BSA and Azide free (AB302505)
  • IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-BCKDHA (phospho S293) antibody [EPR27489-76] - BSA and Azide free (AB302505)

This data was developed using ab302504, the same antibody clone in a different buffer formulation. Immunohistochemical analysis of paraffin-embedded Panel A. Human colon tissue labeling BCKDHA (phospho S293) with ab302504 at 1/500 (0.948 ug/ml) followed by a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101). Mitochondrial staining on human colon without alkaline phosphatase treatment (Panel A). No signal was detected when tissues were treated with alkaline phosphatase (Panel B). The section was incubated with ab302504 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 Rabbit specific IHC polymer detection kit HRP/DAB (ab209101). Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution2) for 20 mins

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-BCKDHA (phospho S293) antibody [EPR27489-76] - BSA and Azide free (AB302505)
  • IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-BCKDHA (phospho S293) antibody [EPR27489-76] - BSA and Azide free (AB302505)

This data was developed using ab302504, the same antibody clone in a different buffer formulation. Immunohistochemical analysis of paraffin-embedded Panel A. Wild-type H tissue labeling BCKDHA (phospho S293) with ab302504 at 1/500 (0.948 ug/ml) followed by a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101). Mitochondrial staining on HEK-293T without alkaline phosphatase treatment (Panel A). No signal was detected when cells were treated with alkaline phosphatase (Panel B). No staining on BCKDHA KO HEK-293T cell pellets (Panel C). The section was incubated with ab302504 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 Rabbit specific IHC polymer detection kit HRP/DAB (ab209101). Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution2) for 20 mins

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-BCKDHA (phospho S293) antibody [EPR27489-76] - BSA and Azide free (AB302505)
  • IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-BCKDHA (phospho S293) antibody [EPR27489-76] - BSA and Azide free (AB302505)

This data was developed using ab302504, the same antibody clone in a different buffer formulation. Immunohistochemical analysis of paraffin-embedded Panel A. Mouse liver tissue labeling BCKDHA (phospho S293) with ab302504 at 1/500 (0.948 ug/ml) followed by a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101). Mitochondrial staining on mouse liver cancer without alkaline phosphatase treatment (Panel A). No signal was detected when tissues were treated with alkaline phosphatase (Panel B). The section was incubated with ab302504 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 Rabbit specific IHC polymer detection kit HRP/DAB (ab209101). Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution2) for 20 mins

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-BCKDHA (phospho S293) antibody [EPR27489-76] - BSA and Azide free (AB302505)
  • IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-BCKDHA (phospho S293) antibody [EPR27489-76] - BSA and Azide free (AB302505)

This data was developed using ab302504, the same antibody clone in a different buffer formulation. Immunohistochemical analysis of paraffin-embedded Panel A. Rat kidney tissue labeling BCKDHA (phospho S293) with ab302504 at 1/500 (0.948 ug/ml) followed by a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101). Mitochondrial staining on rat kidney without alkaline phosphatase treatment (Panel A). No signal was detected when tissues were treated with alkaline phosphatase (Panel B). The section was incubated with ab302504 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 Rabbit specific IHC polymer detection kit HRP/DAB (ab209101). Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution2) for 20 mins

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-BCKDHA (phospho S293) antibody [EPR27489-76] - BSA and Azide free (AB302505)
  • IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-BCKDHA (phospho S293) antibody [EPR27489-76] - BSA and Azide free (AB302505)

This data was developed using ab302504, the same antibody clone in a different buffer formulation. Immunohistochemical analysis of paraffin-embedded Panel A. Mouse kidne tissue labeling BCKDHA (phospho S293) with ab302504 at 1/500 (0.948 ug/ml) followed by a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101). Mitochondrial staining on mouse kidney without alkaline phosphatase treatment (Panel A). No signal was detected when tissues were treated with alkaline phosphatase (Panel B). The section was incubated with ab302504 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 Rabbit specific IHC polymer detection kit HRP/DAB (ab209101). Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution2) for 20 mins

Western blot - Anti-BCKDHA (phospho S293) antibody [EPR27489-76] - BSA and Azide free (AB302505)
  • WB

Supplier Data

Western blot - Anti-BCKDHA (phospho S293) antibody [EPR27489-76] - BSA and Azide free (AB302505)

This data was developed using ab302504, the same antibody clone in a different buffer formulation. Blocking and diluting buffer and concentration : Intercept® (TBS) Blocking Buffer diluted with an equal volume of 0.1% TBS Samples were run on a Bis-Tris gel.Lysates/proteins at 20 µg per lane. Performed under reducing conditions. False colour image of Western blot : Anti- BCKDHA (phospho S293) antibody (ab302504) staining at 1/1000 dilution, shown in green; Mouse anti-GAPDH antibody [6C5] (ab8245) loading control staining at 1/20000 dilution, shown in red.In Western blot, ab302504 was shown to bind specifically to BCKDHA (phospho S293) . A band was observed at 49 kDa in wild-type HEK293T cell lysates with no signal observed at this size in BCKDHA knockout cell line ab266439 (knockout cell lysate ab258324). To generate this image, wild-type and BCKDHA knockout HEK293T cell lysates were analyzed. First, samples were run on an SDS-PAGE gel then transferred onto an immobilon-FL PVDF membrane. Membranes were blocked in in Intercept® (TBS) Blocking Buffer diluted with an equal volume of 0.1% TBS before incubation with primary antibodies overnight at 4 °C. Blots were washed four times in TBS-T, incubated with secondary antibodies for 1 h at room temperature, washed again four times then imaged. Secondary antibodies used were Goat anti-Rabbit IgG H&L (IRDye® 800CW) preabsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye® 680RD) preabsorbed (ab216776) at 1/20000 dilution.

All lanes:

Western blot - Anti-BCKDHA (phospho S293) antibody [EPR27489-76] (<a href='/en-us/products/primary-antibodies/bckdha-phospho-s293-antibody-epr27489-76-ab302504'>ab302504</a>) at 1/1000 dilution

Lane 1:

Wild-type HEK293T (human embryonic kidney epithelial cell), whole cell lysate (untreated membrane) 20 µg

Lane 2:

BCKDHA knockout HEK293T whole cell lysate (untreated membrane) 20 µg

Lane 3:

HeLa (human cervix adenocarcinoma epithelial cell), whole cell lysate (untreated membrane) 20 µg

Lane 4:

C6 (rat glial tumor glial cell), whole cell lysate (untreated membrane) 20 µg

Lane 5:

mIMCD3 (mouse inner medlary collecting duct epithelial cell), whole cell lysate 20 (untreated membrane) 20 µg

Lane 6:

Wild-type HEK293T (human embryonic kidney epithelial cell), whole cell lysate (phosphatase treated membrane) 20 µg

Lane 7:

BCKDHA knockout HEK293T whole cell lysate (phosphatase treated membrane) 20 µg

Lane 8:

HeLa (human cervix adenocarcinoma epithelial cell), whole cell lysate (phosphatase treated membrane) 20 µg

Lane 9:

C6 (rat glial tumor glial cell), whole cell lysate (phosphatase treated membrane) 20 µg

Lane 10:

mIMCD3 (mouse inner medlary collecting duct epithelial cell), whole cell lysate 20 (phosphatase treated membrane) 20 µg

Secondary

Lanes 1 - 10:

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

Lanes 1 - 10:

Western blot - Goat anti-Mouse IgG H&L (IRDye® 680RD) preadsorbed (<a href='/en-us/products/secondary-antibodies/goat-mouse-igg-h-l-irdye-680rd-preadsorbed-ab216776'>ab216776</a>) at 1/10000 dilution

Observed band size: 49 kDa

false

Key facts

Host species

Rabbit

Clonality

Monoclonal

Clone number

EPR27489-76

Isotype

IgG

Carrier free

Yes

Reacts with

Rat, Mouse, Human

Applications

IP, IHC-P, WB

applications

Immunogen

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

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Secondary Antibodies

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Reactivity data

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Product details

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

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 duration
1-2 weeks
Appropriate short-term storage conditions
+4°C
Appropriate long-term storage conditions
+4°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:
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Target data

Together with BCKDHB forms the heterotetrameric E1 subunit of the mitochondrial branched-chain alpha-ketoacid dehydrogenase (BCKD) complex. The BCKD complex catalyzes the multi-step oxidative decarboxylation of alpha-ketoacids derived from the branched-chain amino-acids valine, leucine and isoleucine producing CO2 and acyl-CoA which is subsequently utilized to produce energy. The E1 subunit catalyzes the first step with the decarboxylation of the alpha-ketoacid forming an enzyme-product intermediate. A reductive acylation mediated by the lipoylamide cofactor of E2 extracts the acyl group from the E1 active site for the next step of the reaction.
See full target information BCKDHA phospho S293
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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